KINDAI UNIVERSITY


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NAKAYAMA Takashi

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FacultyDepartment of Pharmacy / Graduate School of Medicine
PositionProfessor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/756-nakayama-takashi.html
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Last Updated :2020/09/04

Education and Career

Education

  •  - 2000 , Osaka University
  •  - 2000 , Osaka University, Graduate School, Division of Pharmaceutical Sciences
  •  - 1992 , Kyoto Pharmaceutical University, Pharmaceutical Sciences
  •  - 1992 , Kyoto Pharmaceutical University, Faculty of Pharmaceutical Science

Academic & Professional Experience

  •   2012 ,  - 現在, Faculty of Pharmacy, Kindai University
  •   2004 ,  - 2012 , Faculty of Medicine, Kindai University
  •   1999 ,  - 2003 , Faculty of Medicine, Kindai University

Research Activities

Research Areas

  • Life sciences, Molecular biology
  • Life sciences, Immunology
  • Life sciences, Virology

Research Interests

  • Chemokine

Published Papers

  • Interstitial-resident memory CD8+ T cells sustain frontline epithelial memory in the lung., Takamura S, Kato S, Motozono C, Shimaoka T, Ueha S, Matsuo K, Miyauchi K, Masumoto T, Katushima A, Nakayama T, Tomura M, Matsushima K, Kubo M, Miyazawa M, Journal of Experimental Medicine, Journal of Experimental Medicine, 216(12), Sep. 2019 , Refereed
  • Epstein-Barr virus-positive pyothorax-associated lymphoma expresses CCL17 and CCL22 chemokines that attract CCR4-expressing regulatory T cells., Higuchi T, Matsuo K, Hashida Y, Kitahata K, Ujihara T, Taniguchi A, Yoshie O, Nakayama T, Daibata M, Cancer letters, Cancer letters, 453, 184 - 192, Jul. 2019 , Refereed
  • Brain regulatory T cells suppress astrogliosis and potentiate neurological recovery., Ito M, Komai K, Mise-Omata S, Iizuka-Koga M, Noguchi Y, Kondo T, Sakai R, Matsuo K, Nakayama T, Yoshie O, Nakatsukasa H, Chikuma S, Shichita T, Yoshimura A, Nature, Nature, 565(7738), 246 - 250, Jan. 2019 , Refereed
  • がんゲノム医療の実装と遺伝性疾患の責任遺伝子探索・創薬から予防までのトータルケアー, 西尾 和人, 田村 和朗, 西郷 和真, 杉浦 麗子, 中山 隆志, 奥野 清隆, 菰池 佳史, 万代 昌紀, 竹村 司, 伊藤 彰彦, 大磯 直毅, 浮田 真沙世, 坂井 和子, 日本遺伝カウンセリング学会誌, 日本遺伝カウンセリング学会誌, 38(2), 35 - 36, May 2017
  • Vaccination with Antigen Combined with alpha beta-ATP as a Vaccine Adjuvant Enhances Antigen-Specific Antibody Production via Dendritic Cell Activation, Kazuhiko Matsuo, Satoshi Nishiuma, Yuta Hasegawa, Fumika Kawabata, Kosuke Kitahata, Takashi Nakayama, BIOLOGICAL & PHARMACEUTICAL BULLETIN, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 39(6), 1073 - 1076, Jun. 2016 , Refereed
    Summary:Adjuvants are required to enhance antigen-specific immune responses by vaccines. Extracellular ATP serves as a danger signal to alert the immune system of tissue damage by acting on P2X and P2Y receptors and triggers the activation of dendritic cells (DCs). Here we investigated the in vivo adjuvant efficacy of alpha,beta-methylene-ATP (alpha beta-ATP), a non-hydrolysable form of ATP. We found that intradermal injection of ovalbumin (OVA), as a model antigen, combined with alpha beta-ATP, as the adjuvant, enhanced OVA-specific immune responses more than OVA alone. Additionally, DCs in the skin of mice injected with OVA and alpha beta-ATP had increased expression of major histocompatibility complex class II and co-stimulator molecules, CD40, CD80, and CD86, suggesting that alpha beta-ATP activated DC. These findings indicate that alpha beta-ATP functions as a potent vaccine adjuvant.
  • CXCL16 suppresses liver metastasis of colorectal cancer by promoting TNF-alpha-induced apoptosis by tumor-associated macrophages, Ji-Ye Kee, Aya Ito, Shozo Hojo, Isaya Hashimoto, Yoshiko Igarashi, Koichi Tsuneyama, Kazuhiro Tsukada, Tatsuro Irimura, Naotoshi Shibahara, Ichiro Takasaki, Akiko Inujima, Takashi Nakayama, Osamu Yoshie, Hiroaki Sakurai, Ikuo Saiki, Keiichi Koizumi, BMC CANCER, BMC CANCER, 14, 949, Dec. 2014 , Refereed
    Summary:Background: Inhibition of metastasis through upregulation of immune surveillance is a major purpose of chemokine gene therapy. In this study, we focused on a membrane-bound chemokine CXCL16, which has shown a correlation with a good prognosis for colorectal cancer (CRC) patients. Methods: We generated a CXCL16-expressing metastatic CRC cell line and identified changes in TNF and apoptosis-related factors. To investigate the effect of CXCL16 on colorectal liver metastasis, we injected SL4-Cont and SL4-CXCL16 cells into intraportal vein in C57BL/6 mice and evaluated the metastasis. Moreover, we analyzed metastatic liver tissues using flow cytometry whether CXCL16 expression regulates the infiltration of M1 macrophages. Results: CXCL16 expression enhanced TNF-alpha-induced apoptosis through activation of PARP and the caspase-3-mediated apoptotic pathway and through inactivation of the NF-kappa B-mediated survival pathway. Several genes were changed by CXCL16 expression, but we focused on IRF8, which is a regulator of apoptosis and the metastatic phenotype. We confirmed CXCL16 expression in SL4-CXCL16 cells and the correlation between CXCL16 and IRF8. Silencing of IRF8 significantly decreased TNF-alpha-induced apoptosis. Liver metastasis of SL4-CXCL16 cells was also inhibited by TNF-alpha-induced apoptosis through the induction of M1 macrophages, which released TNF-alpha. Our findings suggest that the accumulation of M1 macrophages and the enhancement of apoptosis by CXCL16 might be an effective dual approach against CRC liver metastasis. Conclusions: Collectively, this study revealed that CXCL16 regulates immune surveillance and cell signaling. Therefore, we provide the first evidence of CXCL16 serving as an intracellular signaling molecule.
  • Abundant expression of CXCL9 (MIG) by stromal cells that include dendritic cells and accumulation of CXCR3+ T cells in lymphocyte-rich gastric carcinoma, Cancer Sci., Cancer Sci., 217(1), 21 - 31, Jan. 2009
  • Aberrant expression of Fra-2 promotes CCR4 expression and cell proliferation in adult T-cell leukemia, Oncogene, Oncogene, 27(23), 3221 - 3232, May 2008
  • 単純ヘルペスウイルス1型に対するケモカインの抗ウイルス活性の検討, 白根 授美, 中山 隆志, 義江 修, 三島 弘, 下村 嘉一, 日本眼科学会雑誌, 日本眼科学会雑誌, 111(臨増), 157 - 157, Mar. 2007
  • Augmentation of the migratory ability of DC-based vaccine into regional lymph nodes by efficient CCR7 gene transduction, Gene Therapy, Gene Therapy, 12(2), 129 - 139, Jan. 2005
  • IgA抗体産生細胞の腸管ホーミングに関与するケモカイン系の解析, 川崎 ゆり, 稗島 州雄, 花本 仁, 中山 隆志, 長久保 大輔, 金丸 昭久, 義江 修, 近畿大学医学雑誌, 近畿大学医学雑誌, 29(2), 55A - 55A, Oct. 2004
  • Tumor-suppressive activities by chemokines introduced into OV-HM cells using fiber-mutant adenovirus vectors, Pharmazie, Pharmazie, 59(3), 238 - 239, Mar. 2004
  • CCL20のヒト角膜組織における発現誘導とマウスヘルペス性角膜実質炎における役割, 白根 授美, 下村 嘉一, 中山 隆志, 長久保 大輔, 稗島 州雄, 義江 修, 日本眼科学会雑誌, 日本眼科学会雑誌, 108(臨増), 277 - 277, Mar. 2004
  • Transcutaneous immunization with a highly active form of XCL1 as a vaccine adjuvant using a hydrophilic gel patch elicits long-term CD8+ T cell responses, Momo Kamei, Kazuhiko Matsuo, Haruka Imanishi, Yuta Hara, Ying-Shu Quen, Fumio Kamiyama, Naoki Oiso, Akira Kawada, Naoki Okada, Takashi Nakayama, J. Pharmacol. Sci., J. Pharmacol. Sci., 143(3), 182 - 187, Jul. 2020 , Refereed
  • A CCR4 antagonist ameliorates atopic dermatitis-like skin lesions induced by dibutyl phthalate and a hydrogel patch containing ovalbumin., Kazuhiko Matsuo, Shota Hatanaka, Yuta Kimura, Yuta Hara, Keiji Nishiwaki, Ying-Shu Quan, Fumio Kamiyama, Naoki Oiso, Akira Kawada, Kenji Kabashima, Takashi Nakayama, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 109, 1437 - 1444, Jan. 2019 , Refereed
    Summary:CCR4 is a chemokine receptor highly expressed by Th2 cells, and regarded as a potential therapeutic target for atopic dermatitis (AD). CCL17 and CCL22 are the CCR4 ligands, and thymic stromal lymphopoietin (TSLP) is shown to promote the expression of CCL17 and CCL22 by dendritic cells. Here, by using dibutyl phthalate (DBP), a TSLP inducer, and a hydrogel patch as a transcutaneous delivery device for ovalbumin, we developed a novel murine AD model and investigated the effect of Compound 22, a CCR4 antagonist. We first found that the mRNA expression of TSLP together with CCL17 and CCL22 was increased in the skins treated with DBP. Furthermore, the topical application of ovalbumin and DBP efficiently and rapidly induced AD-like skin lesions in BALB/c mice, which were characterized by ear swelling accompanied by infiltration of eosinophils, mast cells, and CCR4-expressing Th2 cells in the skin lesions, and elevated total IgE levels in the sera. Using this AD model, we demonstrated that cutaneous administration of Compound 22 inhibited Th2 cell infiltration and ameliorated the AD-like skin lesions. These results suggest that our AD model could be useful for studying new therapeutic strategies. Collectively, CCR4 antagonists may be a promising approach for treating AD.
  • Ascorbic acid derivative DDH-1 ameliorates psoriasis-like skin lesions in mice by suppressing inflammatory cytokine expression., Kosuke Kitahata, Kazuhiko Matsuo, Yuta Hara, Takanori Naganuma, Naoki Oiso, Akira Kawada, Takashi Nakayama, Journal of pharmacological sciences, Journal of pharmacological sciences, 138(4), 284 - 288, Dec. 2018 , Refereed
    Summary:Psoriasis is a chronic inflammatory skin disease in which inflammatory cytokines play a major role in its pathogenesis. Because DDH-1, a novel amphipathic ascorbic acid derivative, has been recently shown to reduce inflammatory cytokine expression in human keratinocytes in vitro, we investigated its effect on imiquimod-induced psoriasis-like skin lesions in C57BL/6 mice. We first found that IL-1β and TNF-α mRNA expression was significantly decreased in the skin lesions treated with DDH-1. Furthermore, cutaneous administration of DDH-1 ameliorated psoriasis-like skin lesions. These results suggest that DDH-1 may be effective in the prevention and supplemental treatment of psoriasis.
  • CCR4 Is Critically Involved in Skin Allergic Inflammation of BALB/c Mice., Kazuhiko Matsuo, Daisuke Nagakubo, Yuhei Komori, Shun Fujisato, Natsumi Takeda, Mizuki Kitamatsu, Keiji Nishiwaki, Ying-Shu Quan, Fumio Kamiyama, Naoki Oiso, Akira Kawada, Osamu Yoshie, Takashi Nakayama, The Journal of investigative dermatology, The Journal of investigative dermatology, 138(8), 1764 - 1773, Aug. 2018 , Refereed
    Summary:Atopic dermatitis is a chronic inflammatory skin disease involving T-helper (Th) 2 cells, eosinophils, and mast cells. Although CCR4 is a major chemokine receptor expressed on Th2 cells and regarded as a potential therapeutic target for allergic diseases, its role in atopic dermatitis remains unclear. Here, by using a hydrogel patch as a transcutaneous delivery device for ovalbumin (an antigen) and Staphylococcus aureus δ-toxin (a mast cell activator), we efficiently induced acute atopic dermatitis-like skin lesions in BALB/c mice, a strain prone to Th2 responses, which were characterized by increased numbers of eosinophils, mast cells, and CCR4-expressing Th2 cells in the skin lesions; elevated levels of total and ovalbumin-specific IgE in the sera; and increased expression of IL-4, IL-17A, IL-22, CCL17, CCL22, and CCR4 in the skin lesions. Of note, the same model was less efficient in C57BL/6 mice, a strain prone to Th1 responses. Using this atopic dermatitis model in BALB/c mice, we demonstrated that CCR4-deficiency or a CCR4 antagonist ameliorated the allergic responses. Collectively, these results demonstrate that CCR4 plays a pivotal role in skin allergic inflammation of BALB/c mice by recruiting CCR4-expressing Th2 cells and Th17 cells.
  • A CCR4 antagonist enhances DC activation and homing to the regional lymph node and shows potent vaccine adjuvant activity through the inhibition of regulatory T-cell recruitment., Shinya Yamamoto, Kazuhiko Matsuo, Daisuke Nagakubo, Shintaro Higashiyama, Keiji Nishiwaki, Naoki Oiso, Akira Kawada, Osamu Yoshie, Takashi Nakayama, Journal of pharmacological sciences, Journal of pharmacological sciences, 136(3), 165 - 171, Mar. 2018 , Refereed
    Summary:CCR4 is a major chemokine receptor expressed by Treg cells that downregulate immune responses. Here, we investigated the role of CCR4-mediated Treg cell recruitment in antigen-specific immune responses. CCR4-deficient mice immunized intramuscularly with ovalbumin (OVA) showed enhanced OVA-specific IgG responses. Furthermore, intramuscular administration of OVA induced the expression of MDC/CCL22, a ligand for CCR4, in macrophages of the muscle tissues, and enhanced the recruitment of CCR4+ Treg cells in wild-type mice, whereas this recruitment of Treg cells was severely impaired in CCR4-deficient mice. Furthermore, OVA-loaded dendritic cells (DCs) derived from the muscle injection site of CCR4-deficient mice had an upregulated expression of the DC activation marker CD40 and 86, and the lymphoid organ homing receptor CCR7 resulting in an increased number of migratory DCs in the regional lymph node. Compound 22, a CCR4 antagonist, also inhibited the recruitment of Treg cells to the muscle tissue, and further enhanced DC activation and homing to the regional lymph node. Consequently, Compound 22 enhanced OVA-specific IgG responses, and the expression levels of IL-4 and IFN-γ in CD4+ T cells and the levels of IFN-γ in CD8+ T cells. Finally, intramuscular administration of OVA and Compound 22 significantly inhibited the growth of OVA-expressing tumors. Collectively, CCR4 plays a pivotal role in Treg cell recruitment to the muscle tissue, and intramuscular administration of CCR4 antagonists may be a promising approach for enhancing vaccine efficacy.
  • CCL28-Deficient Mice Have Reduced IgA Antibody-Secreting Cells and an Altered Microbiota in the Colon., Kazuhiko Matsuo, Daisuke Nagakubo, Shinya Yamamoto, Akiko Shigeta, Shuta Tomida, Mitsugu Fujita, Takako Hirata, Ikuo Tsunoda, Takashi Nakayama, Osamu Yoshie, Journal of immunology (Baltimore, Md. : 1950), Journal of immunology (Baltimore, Md. : 1950), 200(2), 800 - 809, Jan. 15 2018 , Refereed
    Summary:CCL28 induces the migration of IgA Ab-secreting cells (ASCs) via CCR10 and also displays a potent antimicrobial activity in vitro. To explore the role of CCL28 in vivo, we generated CCL28-deficient mice. The mice exhibited a significant reduction and abnormal distribution of IgA ASCs in the lamina propria of the colon. The concentrations of total and Ag-specific IgA in the fecal extracts of CCL28-deficient mice were also drastically reduced. The average amount of IgA secreted by a single IgA ASC derived from the colon was also substantially reduced in CCL28-deficient mice. Furthermore, CCL28 was found to significantly increase the average amount of IgA secreted by a single IgA ASC derived from the colon in vitro. In contrast, the generation of IgA ASCs in Peyer's and cecal patches was not significantly impaired in CCL28-deficient mice. We also found a relative increase in the Class Bacilli in the fecal extracts of CCL28-deficient mice and demonstrated a potent antimicrobial activity of CCL28 against Bacillus cereus and Enterococcus faecalis, both of which belong to Class Bacilli. Thus, CCL28 may also suppress the outgrowth of some bacterial species by its direct antimicrobial activity. Finally, CCL28-deficient mice exhibited a highly aggravated dextran sodium sulfate-induced colitis that was ameliorated by pretreatment with antibiotics. Collectively, CCL28 plays a pivotal role in the homing, distribution, and function of IgA ASCs in the colon and may also affect the intestinal microbiota through its direct antimicrobial activity.
  • A Highly Active Form of XCL1/Lymphotactin Functions as an Effective Adjuvant to Recruit Cross-Presenting Dendritic Cells for Induction of Effector and Memory CD8+ T Cells., Kazuhiko Matsuo, Kosuke Kitahata, Fumika Kawabata, Momo Kamei, Yuta Hara, Shiki Takamura, Naoki Oiso, Akira Kawada, Osamu Yoshie, Takashi Nakayama, Frontiers in immunology, Frontiers in immunology, 9, 2775 - 2775, 2018 , Refereed
    Summary:The chemokine receptor XCR1 is known to be selectively expressed by cross-presenting dendritic cells (DCs), while its ligand XCL1/lymphotactin is mainly produced by activated CD8+ T cells and natural killer cells. Recent studies have shown that XCL1-antigen fusion proteins efficiently induce CD8+ T cell responses by preferentially delivering antigens to XCR1+ DCs. However, XCL1 per se was found to be a poor adjuvant for induction of CD8+ T cell responses. XCL1 is unique because of its lack of one of the two disulfide bonds commonly conserved in all other chemokines and thus has an unstable structure with a relatively weak chemokine activity. In the present study, we generated a variant form of murine XCL1 termed mXCL1-V21C/A59C that contained a second disulfide bond to stabilize its chemokine structure. We confirmed that mXCL1-V21C/A59C had much more potent chemotactic and calcium mobilization activities than the wild type XCL1 (mXCL1-WT). Intradermal injection of mXCL1-V21C/A59C, but not that of mXCL1-WT, significantly increased the accumulation of XCR1+CD103+ DCs in the injection site, and most of the accumulated XCR1+CD103+ DCs were found to take up co-injected ovalbumin (OVA). Furthermore, recruited XCR1+CD103+ DCs efficiently migrated to the draining lymph nodes and stayed for a prolonged period of time. Consequently, mXCL1-V21C/A59C strongly induced OVA-specific CD8+ T cells. The combination of OVA and mXCL1-V21C/A59C well protected mice from E.G7-OVA tumor growth in both prophylactic and therapeutic protocols. Finally, memory CTL responses were efficiently induced in mice immunized with OVA and mXCL1-V21C/A59C. Although intradermal injection of OVA and polyinosinic-polycytidylic acid (poly(I:C)) as an adjuvant also induced CD8+ T cell responses to OVA, poly (I:C) poorly recruited XCR1+CD103+ DCs in the injection site and failed to induce significant memory CTL responses to OVA. Collectively, our findings demonstrate that a highly active form of XCL1 is a promising vaccine adjuvant for cross-presenting DCs to induce antigen-specific effector and memory CD8+ T cells.
  • Impaired lymphocyte trafficking in mice deficient in the kinase activity of PKN1, Rana Mashud, Akira Nomachi, Akihide Hayakawa, Koji Kubouchi, Sally Danno, Takako Hirata, Kazuhiko Matsuo, Takashi Nakayama, Ryosuke Satoh, Reiko Sugiura, Manabu Abe, Kenji Sakimura, Shigeharu Wakana, Hiroyuki Ohsaki, Shingo Kamoshida, Hideyuki Mukai, SCIENTIFIC REPORTS, SCIENTIFIC REPORTS, 7(1), 7663, Aug. 2017 , Refereed
    Summary:Knock-in mice lacking PKN1 kinase activity were generated by introducing a T778A point mutation in the catalytic domain. PKN1[T778A] mutant mice developed to adulthood without apparent external abnormalities, but exhibited lower T and B lymphocyte counts in the peripheral blood than those of wild-type (WT) mice. T and B cell development proceeded in an apparently normal fashion in bone marrow and thymus of PKN1[T778A] mice, however, the number of T and B cell counts were significantly higher in the lymph nodes and spleen of mutant mice in those of WT mice. After transfusion into WT recipients, EGFP-labelled PKN1[T778A] donor lymphocytes were significantly less abundant in the peripheral circulation and more abundant in the spleen and lymph nodes of recipient mice compared with EGFP-labelled WT donor lymphocytes, likely reflecting lymphocyte sequestration in the spleen and lymph nodes in a cell-autonomous fashion. PKN1[T778A] lymphocytes showed significantly lower chemotaxis towards chemokines and sphingosine 1-phosphate (S1P) than WT cells in vitro. The biggest migration defect was observed in response to S1P, which is essential for lymphocyte egress from secondary lymphoid organs. These results reveal a novel role of PKN1 in lymphocyte migration and localization.
  • Contributions of MET activation to BCR-ABL1 tyrosine kinase inhibitor resistance in chronic myeloid leukemia cells., Masanobu Tsubaki, Tomoya Takeda, Toshiki Kino, Kazuko Sakai, Tatsuki Itoh, Motohiro Imano, Takashi Nakayama, Kazuto Nishio, Takao Satou, Shozo Nishida, Oncotarget, Oncotarget, 8(24), 38717 - 38730, Jun. 13 2017 , Refereed
    Summary:Resistance to the breakpoint cluster region-abelson 1 (BCR-ABL1) tyrosine kinase inhibitor (TKI) imatinib poses a major problem when treating chronic myeloid leukemia (CML). Imatinib resistance often results from a secondary mutation in BCR-ABL1. However, in the absence of a mutation in BCR-ABL1, the basis of BCR-ABL1-independent resistance must be elucidated. To gain insight into the mechanisms of BCR-ABL1-independent imatinib resistance, we performed an array-based comparative genomic hybridization. We identified various resistance-related genes, and focused on MET. Treatment with a MET inhibitor resensitized K562/IR cells to BCR-ABL1 TKIs. Combined treatment of K562/IR cells with imatinib and a MET inhibitor suppressed extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK) activation, but did not affect AKT activation. Our findings implicate the MET/ERK and MET/JNK pathways in conferring resistance to imatinib, providing new insights into the mechanisms of BCR-ABL1 TKI resistance in CML.
  • CCR4 is critically involved in effective antitumor immunity in mice bearing intradermal B16 melanoma., Kazuhiko Matsuo, Tatsuki Itoh, Atsushi Koyama, Reira Imamura, Shiori Kawai, Keiji Nishiwaki, Naoki Oiso, Akira Kawada, Osamu Yoshie, Takashi Nakayama, Cancer letters, Cancer letters, 378(1), 16 - 22, Aug. 01 2016 , Refereed
    Summary:CCR4 is a major chemokine receptor expressed by Treg cells and Th17 cells. While Treg cells are known to suppress antitumor immunity, Th17 cells have recently been shown to enhance the induction of antitumor cytotoxic T lymphocytes. Here, CCR4-deficient mice displayed enhanced tumor growth upon intradermal inoculation of B16-F10 melanoma cells. In CCR4-deficient mice, while IFN-γ+CD8+ effector T cells were decreased in tumor sites, IFN-γ+CD8+ T cells and Th17 cells were decreased in regional lymph nodes. In wild-type mice, CD4+IL-17A+ cells, which were identified as CCR4+CD44+ memory Th17, were found to be clustered around dendritic cells expressing MDC/CCL22, a ligand for CCR4, in regional lymph nodes. Compound 22, a CCR4 antagonist, also enhanced tumor growth and decreased Th17 cells in regional lymph nodes in tumor-bearing mice treated with Dacarbazine. In contrast, CCR6 deficiency did not affect the tumor growth and the numbers of Th17 cells in regional lymph nodes. These findings indicate that CCR4 is critically involved in regional lymph node DC-Th17 cell interactions that are necessary for Th17 cell-mediated induction of antitumor CD8+ effector T cells in mice bearing B16 melanoma.
  • Neolignans from the Arils of Myristica fragrans as Potent Antagonists of CC Chemokine Receptor 3, Toshio Morikawa, Ikuko Hachiman, Kazuhiko Matsuo, Eriko Nishida, Kiyofumi Ninomiya, Takao Hayakawa, Osamu Yoshie, Osamu Muraoka, Takashi Nakayama, JOURNAL OF NATURAL PRODUCTS, JOURNAL OF NATURAL PRODUCTS, 79(8), 2005 - 2013, Aug. 2016 , Refereed
    Summary:CC chemokine receptor 3 (CCR3) is expressed selectively in eosinophils, basophils, and some Th2 cells and plays a major role in allergic diseases. A methanol extract from the arils of Myristica fragrans inhibited CC chemokine ligand 11-induced chemotaxis in CCR3-expressing L1.2 cells at 100 mu g/mL. From this extract, eight new neolignans, maceneolignans A-H (1-8), were isolated, and their stereostructures were elucidated from their spectroscopic values and chemical properties. Of those constituents, compounds 1, 4, 6, and 8 and (+)-erythro-(7S,8R)-Delta(8)'-7-hydroxy-3,4-methylenedioxy-3,5'-dimethoxy-8-O-4'-neolignan (11), (-)- (8R)-Delta(8,)-3,4-methylene dioxy-3',5-dimethoxy-8-O-4'-neolignan (17), (+)-licarin A (20), nectandrin B (25), verrucosin (26), and myristicin (27) inhibited CCR3-mediated chemotaxis at, a concentration of 1 mu M. Among them, 1 (EC50 1.6 mu M), 6 (1.5 mu M), and 8 (1.4 mu M) showed relatively strong activities, which were comparable to that of a synthetic CCR3 selective antagonist, SB328437 (0.78 mu M).
  • C-C chemokine receptor type 4 antagonist Compound 22 ameliorates experimental autoimmune encephalomyelitis, Kota Moriguchi, Katsuichi Miyamoto, Noriko Tanaka, Rino Ueno, Takashi Nakayama, Osamu Yoshie, Susumu Kusunoki, JOURNAL OF NEUROIMMUNOLOGY, JOURNAL OF NEUROIMMUNOLOGY, 291, 54 - 58, Feb. 2016 , Refereed
    Summary:Chemokines and chemokine receptors play important roles in the immune response. We previously reported the pathogenic role of C-C chemokine receptor type 4 (CCR4) in experimental autoimmune encephalomyelitis (EAE). Here, we examined whether CCR4 antagonism modulates the disease course of EAE. Wild-type and CCR4-knockout mice were induced EAE and were administered Compound 22, an antagonist of CCR4. Compound 22 significantly ameliorated the severity of EAE in wild-type mice, but not in the CCR4-knockout mice. Compound 22 inhibited Th1 and Th17 polarization of antigen-induced T-cell responses. Therefore, CCR4 antagonists might be potential therapeutic agents for multiple sclerosis. (C) 2015 Elsevier B.V. All rights reserved.
  • Efficient Use of a Crude Drug/Herb Library Reveals Ephedra Herb As a Specific Antagonist for TH2-Specific Chemokine Receptors CCR3, CCR4, and CCR8., Kazuhiko Matsuo, Keiichi Koizumi, Mitsugu Fujita, Toshio Morikawa, Michiko Jo, Naotoshi Shibahara, Ikuo Saiki, Osamu Yoshie, Takashi Nakayama, Frontiers in cell and developmental biology, Frontiers in cell and developmental biology, 4, 54 - 54, 2016 , Refereed
    Summary:Chemokine receptors CCR3 and CCR4 are preferentially expressed by TH2 cells, mast cells, and/or eosinophils, all of which are involved in the pathogenesis of allergic diseases. Therefore, CCR3 and CCR4 have long been highlighted as potent therapeutic targets for allergic diseases. Japanese traditional herbal medicine Kampo consists of multiple crude drugs/herbs, which further consist of numerous chemical substances. Recent studies have demonstrated that such chemical substances appear to promising sources in the development of novel therapeutic agents. Based on these findings, we hypothesize that Kampo-related crude drugs/herbs would contain chemical substances that inhibit the cell migration mediated by CCR3 and/or CCR4. To test this hypothesis, we screened 80 crude drugs/herbs to identify candidate substances using chemotaxis assay. Among those tested, Ephedra Herb inhibited the chemotaxis mediated by both CCR3 and CCR4, Cornus Fruit inhibited that mediated by CCR3, and Rhubarb inhibited that mediated by CCR4. Furthermore, Ephedra Herb specifically inhibited the chemotaxis mediated by not only CCR3 and CCR4 but CCR8, all of which are selectively expressed by TH2 cells. This result led us to speculate that ephedrine, a major component of Ephedra Herb, would play a central role in the inhibitory effects on the chemotaxis mediated by CCR3, CCR4, and CCR8. However, ephedrine exhibited little effects on the chemotaxis. Therefore, we fractionated Ephedra Herb into four subfractions and examined the inhibitory effects of each subfraction. As the results, ethyl acetate-insoluble fraction exhibited the inhibitory effects on chemotaxis and calcium mobilization mediated by CCR3 and CCR4 most significantly. In contrast, chloroform-soluble fraction exhibited a weak inhibitory effect on the chemotaxis mediated by CCR8. Furthermore, maoto, one of the Kampo formulations containing Ephedra Herb, exhibited the inhibitory effects on the chemotaxis mediated by CCR3, CCR4, and CCR8. Taken together, our data suggest that these crude drugs/herbs might be useful sources to develop new drugs targeting TH2-mediated allergic diseases.
  • Role of chemokine CX3CL1 in progression of multiple myeloma via CX3CR1 in bone microenvironments, Akinori Wada, Aya Ito, Hirofumi Iitsuka, Koichi Tsuneyama, Takayoshi Miyazono, Jun Murakami, Naotoshi Shibahara, Hiroaki Sakurai, Ikuo Saiki, Takashi Nakayama, Osamu Yoshie, Keiichi Koizumi, Toshiro Sugiyama, ONCOLOGY REPORTS, ONCOLOGY REPORTS, 33(6), 2935 - 2939, Jun. 2015 , Refereed
    Summary:Several chemokines/chemokine receptors such as CXCL12, CCL3, CXCR4 and CCR1 attract multiple myelomas to specific microenvironments. In the present study, we investigated whether the CX3CL1/CX3CR1 axis is involved in the interaction of the multiple myeloma cells with their microenvironment. The expression of CX3CR1 (also known as fractalkine) was detected in three of the seven human myeloma cell lines. CX3CL1-induced phosphorylation of Akt and ERK1/2 was detected in the CX3CR1-positive cell lines, but not in the CX3CR1-negative cell lines. In addition, CX3CL1-induced cell adhesion to fibronectin and vascular cell adhesion molecule-1 (VCAM-1) in the human myeloma RPMI-8226 cell line. We also investigated whether a relationship existed between myeloma cells and osteoclasts that may function via the CX3CL1/CX3CR1 axis. Conditioned medium from CX3CL1-stimulated RPMI-8226 cells drastically increased the osteoclast differentiation. Collectively, the results from the present study support the concept of the CX3CL1-mediated activation of the progression of the multiple myeloma via CX3CR1. Thus, CX3CR1 may represent a potential therapeutic target for the treatment of multiple myeloma in a bone microenvironment.
  • Structural and agonist properties of XCL2, the other member of the C-chemokine subfamily, Jamie C. Fox, Takashi Nakayama, Robert C. Tyler, Tara L. Sander, Osamu Yoshie, Brian F. Volkman, CYTOKINE, CYTOKINE, 71(2), 302 - 311, Feb. 2015 , Refereed
    Summary:Known for its unusual metamorphic native state structure, XCL1 has been the focus of most efforts to elucidate the structural, functional, and physiological properties of chemokines in the C subfamily. By comparison, its closely related paralog XCL2 remains virtually uncharacterized. Based on the importance of the chemokine N-terminus in receptor activation, it was hypothesized that two amino acid differences in XCL2 would alter its agonist activity relative to XCL1 for their shared receptor XCR1. This present study reveals several properties of XCL2 that were unexamined until now. Structurally, XCL1 and XCL2 are very similar, exchanging between the monomeric chemokine fold and an unrelated dimeric state under physiological NaCl and temperature conditions. Ca2+ flux, chemotaxis, and heparin binding assays showed that the monomer form of XCL2 is responsible for G protein-coupled receptor activation while the dimeric form is important for GAG binding. Despite their high structural similarity, XCL2 displays a slightly higher affinity for heparin than XCL1. Because their in vitro functional profiles are virtually identical, distinct physiological roles for XCL1 and XCL2 are probably encoded at the level of expression. (C) 2014 Elsevier Ltd. All rights reserved.
  • A pleomorphic carcinoma of the lung producing multiple cytokines and forming a rapidly progressive mass-like opacity, Masataka Matsumoto, Takashi Nakayama, Daiki Inoue, Kazufumi Takamatsu, Ryo Itotani, Manabu Ishitoko, Shinko Suzuki, Minoru Sakuramoto, Yoshiaki Yuba, Osamu Yoshie, Masaya Takemura, Motonari Fukui, BMC CANCER, BMC CANCER, 14, 588, Aug. 2014 , Refereed
    Summary:Background: Lung cancer cells have been reported to produce cytokines, resulting in systemic reactions. There have been few reports showing that these cytokines induced the formation of an inflammatory mass around lung cancers. Case presentation: We encountered a patient with a pleomorphic carcinoma of the lung. This tumor produced interleukin (IL)-8, granulocyte colony-stimulating factor and IL-6, which in turn recruited inflammatory cells, such as CD8 positive lymphocytes, around the tumor, resulting in a rapidly growing tumor shadow. Conclusion: 18 F-fluoro-deoxy-glucose positron emission tomography, in addition to a conventional radiological approach such as computed tomography, may detect immunological responses around a tumor.
  • Necrobiosis lipoidica with infiltration of Th17 cells into vascular lesions, Maiko Kato, Naoki Oiso, Tatsuki Itoh, Masako Sato, Kazuhiko Matsuo, Takashi Nakayama, Takao Satou, Akira Kawada, JOURNAL OF DERMATOLOGY, JOURNAL OF DERMATOLOGY, 41(5), 459 - 461, May 2014 , Refereed
  • Neuroprotective effect of (-)-epigallocatechin-3-gallate in rats when administered pre- or post-traumatic brain injury, Tatsuki Itoh, Masaki Tabuchi, Nobuyuki Mizuguchi, Motohiro Imano, Masahiro Tsubaki, Shozo Nishida, Shigeo Hashimoto, Kazuhiko Matsuo, Takashi Nakayama, Akihiko Ito, Hiroshi Munakata, Takao Satou, JOURNAL OF NEURAL TRANSMISSION, JOURNAL OF NEURAL TRANSMISSION, 120(5), 767 - 783, May 2013 , Refereed
    Summary:Our previous study indicated that consuming (-)-epigallocatechin gallate (EGCG) before or after traumatic brain injury (TBI) eliminated free radical generation in rats, resulting in inhibition of neuronal degeneration and apoptotic death, and improvement of cognitive impairment. Here we investigated the effects of administering EGCG at various times pre- and post-TBI on cerebral function and morphology. Wistar rats were divided into five groups and were allowed access to (1) normal drinking water, (2) EGCG pre-TBI, (3) EGCG pre- and post-TBI, (4) EGCG post-TBI, and (5) sham-operated group with access to normal drinking water. TBI was induced with a pneumatic controlled injury device at 10 weeks of age. Immunohistochemistry and lipid peroxidation studies revealed that at 1, 3, and 7 days post-TBI, the number of 8-Hydroxy-2'-deoxyguanosine-, 4-Hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells, and levels of malondialdehyde around the damaged area were significantly decreased in all EGCG treatment groups compared with the water group (P < 0.05). Although there was a significant increase in the number of surviving neurons after TBI in each EGCG treatment group compared with the water group (P < 0.05), significant improvement of cognitive impairment after TBI was only observed in the groups with continuous and post-TBI access to EGCG (P < 0.05). These results indicate that EGCG inhibits free radical-induced neuronal degeneration and apoptotic death around the area damaged by TBI. Importantly, continuous and post-TBI access to EGCG improved cerebral function following TBI. In summary, consumption of green tea may be an effective therapy for TBI patients.
  • SOX4 is a direct target gene of FRA-2 and induces expression of HDAC8 in adult T-cell leukemia/lymphoma, Tomonori Higuchi, Takashi Nakayama, Tokuzo Arao, Kazuto Nishio, Osamu Yoshie, BLOOD, BLOOD, 121(18), 3640 - 3649, May 2013 , Refereed
    Summary:Previously, we have shown that an AP-1 family member, FRA-2, is constitutively expressed in adult T-cell leukemia/lymphoma (ATL) and, together with JUND, upregulates CCR4 and promotes ATL cell growth. Among the identified potential target genes of FRA-2/JUND was SOX4. Here, we examine the expression and function of SOX4 in ATL. SOX4 was indeed consistently expressed in primary ATL cells. FRA-2/JUND efficiently activated the SOX4 promoter via an AP-1 site. Knockdown of SOX4 expression by small interfering RNA (siRNA) strongly suppressed cell growth of ATL cell lines. Microarray analyses revealed that SOX4 knockdown reduced the expression of genes such as germinal center kinase related (GCKR), NAK-associated protein 1 (NAP1), and histone deacetylase 8 (HDAC8). We confirmed consistent expression of GCKR, NAP1, and HDAC8 in primary ATL cells. We also showed direct activation of the HDAC8 promoter by SOX4. Furthermore, siRNA knockdown of GCKR, NAP1, and HDAC8 each significantly suppressed cell growth of ATL cell lines. Taken together, we have revealed an important oncogenic cascade involving FRA-2/JUND and SOX4 in ATL, which leads to the expression of genes such as GCKR, NAP1, and HDAC8.
  • Appearance of neural stem cells around the damaged area following traumatic brain injury in aged rats, Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Takashi Nakayama, Nobuyuki Mizuguchi, Shigeaki Yamanaka, Masaki Tabuchi, Hiroshi Munakata, Shigeo Hashimoto, Akihiko Ito, Takao Satou, JOURNAL OF NEURAL TRANSMISSION, JOURNAL OF NEURAL TRANSMISSION, 120(3), 361 - 374, Mar. 2013 , Refereed
    Summary:We have previously reported free radical production after traumatic brain injury (TBI), which induces neural stem cell (NSC) degeneration and death. However, the effects of aging on NSC proliferation around the damaged area following TBI have not been investigated. Therefore, in this study, we used 10-week (young group) and 24-month-old (aged group) rat TBI models to investigate the effects of aging on NSC proliferation around damaged tissue using immunohistochemical and ex vivo techniques. Young and aged rats received TBI. At 1, 3 and 7 days after TBI, immunohistochemical and lipid peroxidation studies were performed. Immunohistochemistry revealed that the number of nestin-positive cells around the damaged area after TBI in the aged group decreased significantly when compared with those in the young group (P < 0.01). However, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells and the level of peroxidation around the damaged area after TBI significantly increased in the aged group, compared with those in the young group (P < 0.01). Furthermore, almost all ssDNA-positive cells in young and aged groups co-localized with NeuN and nestin staining. Ex vivo studies revealed that neurospheres, which differentiated into neurons and glia in culture, could only be isolated from injured brain tissue in young and aged groups at 3 days after TBI. These results indicate that, although there were fewer NSCs that have the potential to differentiate into neurons and glia, these NSCs escaped free radical-induced degeneration around the damaged area after TBI in the aged rat brain.
  • Chemokine CXCL16 suppresses liver metastasis of colorectal cancer via augmentation of tumor-infiltrating natural killer T cells in a murine model, Ji-Ye Kee, Aya Ito, Shozo Hojo, Isaya Hashimoto, Yoshiko Igarashi, Kazuhiro Tsukada, Tatsuro Irimura, Naotoshi Shibahara, Takashi Nakayama, Osamu Yoshie, Hiroaki Sakurai, Ikuo Saiki, Keiichi Koizumi, Oncology Reports, Oncology Reports, 29(3), 975 - 982, Mar. 2013 , Refereed
    Summary:Colorectal cancer (CRC) is a typical lifestyle-related disease, and it metastasizes mostly to the liver. It is important to understand the molecular mechanisms of CRC metastasis in order to design new and effective treatments for CRC patients. Chemokines are known to have antitumor effects as their chemoattractant properties stimulate the accumulation of infiltrating immune cells (TILs) in tumors. Chemokine (C-X-C motif) ligand 16 (CXCL16), also known as SR-PSOX, is a unique membrane-bound chemokine that induces the expression of its specific receptor CXCR6. We previously reported that the expression of CXCL16 by cancer cells enhances the recruitment of TILs, thereby improving the prognosis of CRC. It has since been reported that CXCL16/CXCR6 expression is involved in the metastasis of various types of cancer. However, there is no report of the association between CXCL16 expression and liver metastasis in CRC. In this study, we investigated the role of cancer-derived CXCL16 and the possibility of gene therapy using CXCL16. Therefore, we examined the metastasis of colon 38 SL4 cells to the liver in an experimental model. Following injection of cancer cells into the intraportal vein, CXCL16-expressing CRC cells drastically inhibited liver metastasis. We also found that CD8 T cells and natural killer T (NKT) cells, known as CXCR6-expressing cells, increased in CXCL16-expressing metastatic tissue. Collectively, the inhibitory effect on metastasis to the liver by CXCL16 was observed in NKT cell-depleted mice but not in CD8 T cell-depleted mice. These results demonstrate the inhibitory effect of CXCL16 on liver metastasis via NKT cells in CRC.
  • Expression and Function of FRA2/JUND in Cutaneous T-Cell Lymphomas, Takashi Nakayama, Tomonori Higuchi, Naoki Oiso, Akira Kawada, Osamu Yoshie, ANTICANCER RESEARCH, ANTICANCER RESEARCH, 32(4), 1367 - 1373, Apr. 2012 , Refereed
    Summary:Adult T-cell leukemia/lymphoma (ATLL) and cutaneous T-cell lymphomas (CTCLs) are known to frequently express CC chemokine receptor 4 (CCR4). Previously, we investigated the transcriptional control of CCR4 expression in ATLL and have found that an activating protein 1 (AP1) family member, FBJ murine osteosarcoma viral oncogene homolog (FOS)-related antigen 2 (FRA2), is consistently expressed at high levels in ATLL and, together with v-JUN avian sarcoma virus 17 oncogene homolog D (JUND), up-regulates the expression of CCR4 as well as that of several proto-oncogenes such as v-MYB myeloblastosis viral oncogene homolog (MYB), murine double minute 2 homolog (MDM2), and B-cell lymphoma 6 (BCL6). Here, we examined the expression of these genes in clinical samples of CTCLs. We detected the transcripts of FRA2, JUND, CCR4, MYB, MDM2, and BCL6 at high levels in CTCL skin lesions. Except for BCL6, we confirmed protein expression of FRA2, JUND, CCR4, MYB, and MDM2 in CTCL skin lesions. Furthermore, siRNA-mediated knockdown of FRA2 or JUND suppressed cell growth and the expression of CCR4, MYB, MDM2, and BCL6 in CTCL cell lines. Our results, thus, demonstrate the presence of a common oncogenic cascade initiated by FRA2/JUND in CCR4-expressing mature T-cell malignancies such as ATLL and CTCLs.
  • In situ expression of the CCL20-CCR6 axis in lymphocyte-rich gastric cancer and its potential role in the formation of lymphoid stroma, Haruo Ohtani, Takashi Nakayama, Osamu Yoshie, PATHOLOGY INTERNATIONAL, PATHOLOGY INTERNATIONAL, 61(11), 645 - 651, Nov. 2011 , Refereed
    Summary:Lymphocyte-rich gastric cancer (Ly-rich GC) is characterized by lymphoid stroma. To understand its formation, we studied the expression of a chemokine ligand (CCL)20 and its receptor CCR6 in 36 and 37 cases of Ly-rich- and conventional GC, respectively. Lymphoid tissues in the alimentary tract were studied in parallel. By quantitative polymerase chain reaction, Ly-rich GC contained CCL20 and CCR6 mRNAs at higher levels than conventional GC. By immunohistochemistry, CCL20 was expressed by cancer cells more frequently in Ly-rich GC than in conventional GC. This was comparable with its expression in epithelial cells of the alimentary tract lymphoid tissues. CCR6 was mostly expressed by dendritic cells (DC) and B cells in Ly-rich GC, which was also comparable with its expression in the alimentary tract lymphoid tissues. Cancer cells also expressed CCR6. However, its expression did not differ between Ly-rich- and conventional GC, nor was it related to the stage of cancer. Given that the CCL20-CCR6 axis is involved in the formation of alimentary tract lymphoid tissue, the similarity between the lymphoid stroma of Ly-rich GC and the alimentary tract lymphoid tissues supports the notion that it plays a significant role in the formation of lymphoid stroma in Ly-rich GC.
  • Eotaxin-3/CC Chemokine Ligand 26 Is a Functional Ligand for CX3CR1, Takashi Nakayama, Yoshiko Watanabe, Naoki Oiso, Tomonori Higuchi, Akiko Shigeta, Nobuyuki Mizuguchi, Fuminori Katou, Kenji Hashimoto, Akira Kawada, Osamu Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 185(11), 6472 - 6479, Dec. 2010 , Refereed
    Summary:Eotaxin-3/CCL26 is a functional ligand for CCR3 and abundantly produced by IL-4-/IL-13-stimulated vascular endothelial cells. CCL26 also functions as a natural antagonist for CCR1, CCR2, and CCR5. In this study, we report that CCL26 is yet a functional ligand for CX3CR1, the receptor for fractalkine/CX3CL1, which is expressed by CD16(+) NK cells, cytotoxic effector CD8(+) T cells, and CD14(low)CD16(high) monocytes. Albeit at relatively high concentrations, CCL26 induced calcium flux and chemotaxis in mouse L1.2 cells expressing human CX3CR1 but not mouse CX3CR1 and competed with CX3CL1 for binding to CX3CR1. In chemotaxis assays using human PBMCs, CCL26 attracted not only eosinophils but also CD16(+) NK cells, CD45RA(+)CD27(-)CD8(+) T cells, and CD14(low)CD16(high) monocytes. Intraperitoneal injection of CCL26 into mice rapidly recruited mouse eosinophils and intravenously transferred human CD16(+) NK cells into the peritoneal cavity. IL-4-stimulated HUVECs produced CCL26 and efficiently induced adhesion of cells expressing CX3CR1. Real-time PCR showed that skin lesions of psoriasis consistently contained CX3CL1 mRNA but not CCL26 mRNA, whereas those of atopic dermatitis contained CCL26 mRNA in all samples but CX3CL1 mRNA in only about half of the samples. Nevertheless, the skin lesions from both diseases consistently contained CX3CR1 mRNA at high levels. Thus, CCL26 may be partly responsible for the recruitment of cells expressing CX3CR1 in atopic dermatitis particularly when the expression of CX3CL1 is low. Collectively, CCL26 is another agonist for CX3CR1 and may play a dual role in allergic diseases by attracting eosinophils via CCR3 and killer lymphocytes and resident monocytes via CX3CR1. The Journal of Immunology, 2010, 185: 6472-6479.
  • Tumor-infiltrating lymphocytes, particularly the balance between CD8(+) T cells and CCR4(+) regulatory T cells, affect the survival of patients with oral squamous cell carcinoma, Yoshiko Watanabe, Fuminori Katou, Haruo Ohtani, Takashi Nakayama, Osamu Yoshie, Kenji Hashimoto, ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTOLOGY, ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTOLOGY, 109(5), 744 - 752, May 2010 , Refereed
    Summary:Objective. The objective of this study was to clarify the prognostic significance of tumor-infiltrating lymphocytes (TILs) in oral squamous cell carcinoma (OSCC); the present study analyzed various TIL-related parameters. Study design. Immunohistochemistry was performed in 87 patients with OSCC for the following TIL-related parameters: nest-CD8(+) T cells, stromal CD8(+) T cells, CD4(+) T cells, total regulatory T cells (Tregs), CCR4(+) Tregs, ratio of nest CD8(+) T cells/CCR4(+) Tregs, and ratio of stromal CD8(+) T cells/CCR4(+) Tregs. Results. In univariate analyses, the following parameters were associated with decreased survival: few nest-and stromal CD8(+) T cells and more stromal CCR4(+) Tregs, but not total Tregs. Low ratios of nest and stromal CD8(+) T cell/CCR4(+) Treg were associated with worse survival. In multivariate analysis, the stromal CD8(+) T cell/CCR4(+) Treg ratio was an independent prognostic factor. Conclusion. Host immune responses in the stroma of OSCC affect the survival of the patients. The in situ balance between effector T cells and regulatory T cells is the most important factor predicting survival. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 744-752)
  • Role of CX3CL1/Fractalkine in Osteoclast Differentiation and Bone Resorption, Keiichi Koizumi, Yurika Saitoh, Takayuki Minami, Nobuhiro Takeno, Koichi Tsuneyama, Tatsuro Miyahara, Takashi Nakayama, Hiroaki Sakurai, Yasuo Takano, Miyuki Nishimura, Toshio Imai, Osamu Yoshie, Ikuo Saiki, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 183(12), 7825 - 7831, Dec. 2009 , Refereed
    Summary:The recruitment of osteoclast precursors toward osteoblasts and subsequent cell-cell interactions are critical for osteoclast differentiation. Chemokines are known to regulate cell migration and adhesion. CX3CL1 (also called fractalkine) is a unique membrane-bound chemokine, that has dual functions for cells expressing its receptor CX3CR1: a potent chemotactic factor in its soluble form and a type of efficient cell adhesion molecule in its membrane-bound form. In this paper, we demonstrate a novel role of CX3CL1 in osteoblast-induced osteoclast differentiation. We found that osteoclast precursors selectively expressed CX3CR1, whereas CX3CL1 is expressed by osteoblasts. We confirmed that soluble CX3CL1 induced migration of bone marrow cells containing osteoclast precursors, whereas immobilized CX3CL1 mediated firm adhesion of osteoclast precursors. Furthermore, a blocking mAb against CX3CL1 efficiently inhibited osteoclast differentiation in mouse bone marrow cells cocultured with osteoblasts. Anti-CX3CL1 also significantly suppressed bone resorption in neonatal mice by reducing the number of bone-resorbing mature osteoclasts. Collectively, CX3CL1 expressed by osteoblasts plays an important role in osteoclast differentiation, possibly through its dual functions as a chemotactic factor and adhesion molecule for osteoclast precursors expressing CX3CR1. The CX3CL1-CX3CR1 axis may be a novel target for the therapeutic intervention of bone resorbing diseases such as rheumatoid arthritis, osteoporosis, and cancer bone metastasis. The Journal of Immunology, 2009, 183: 7825-7831.
  • CXCR7 is inducible by HTLV-1 Tax and promotes growth and survival of HTLV-1-infected T cells, Zhe Jin, Daisuke Nagakubo, Aiko-Konno Shirakawa, Takashi Nakayama, Akiko Shigeta, Kunio Hieshima, Yasuaki Yamada, Osamu Yoshie, INTERNATIONAL JOURNAL OF CANCER, INTERNATIONAL JOURNAL OF CANCER, 125(9), 2229 - 2235, Nov. 2009 , Refereed
    Summary:Human T-lymphotropic virus type 1 (HTLV-1), the etiological agent of adult T-cell leukemia (ATL), encodes the potent transcriptional activator Tax, which is required for HTLV-1-induced immortalization of T cells. CXCR7 is an atypical chemokine receptor frequently expressed by tumor cells and known to promote cell growth and survival. We found that HTLV-1-immortalized T cells expressing Tax consistently expressed CXCR7. Induction of Tax in JPX-9 upregulated CXCR7. Wild-type Tax efficiently activated the CXCR7 promoter via a proximal NF-kappa B site, while a mutant Tax selectively defective in NF-kappa B activation did not. CCX754, a synthetic CXCR7 antagonist, inhibited cell growth and increased apoptosis of HTLV-1-immortalized T cells. Knockdown of CXCR7 by small interfering RNA also reduced cell growth. Stable expression of CXCR7 in a CXCR7-negative ATL cell line promoted cell growth and survival. Taken together, CXCR7 is inducible by Tax and may play an important role in HTLV-1-induced immortalization of T cells by promoting growth and survival of HTLV-1-infected T cells. (C) 2009 UICC
  • NF-kappa B-mediated activation of the chemokine CCL22 by the product of the human cytomegalovirus gene UL144 escapes regulation by viral IE86, Emma Poole, Elizabeth Atkins, Takashi Nakayama, Osamu Yoshie, Ian Groves, Antonio Alcami, John Sinclair, JOURNAL OF VIROLOGY, JOURNAL OF VIROLOGY, 82(9), 4250 - 4256, May 2008 , Refereed
    Summary:The product of the human cytomegalovirus (HCMV) gene UL144, expressed at early times postinfection, is located in the UL/b' region of the viral genome and is related to members of the tumor necrosis factor receptor superfamily, but it does not bind tumor necrosis factor superfamily ligands. However, UL144 does activate NF-kappa B, resulting in NF-kappa B-mediated activation of the cellular chemokine CCL22. Consistent with this finding, isolates of HCW lacking the UL/b' region show no such activation of CCL22. Recently, it has been suggested that activation of NF-kappa B is repressed by the product of the viral gene IE86: IE86 appears to block NF-kappa B binding to DNA but not nuclear translocation of NF-kappa B. Intriguingly, IE86 is detectable throughout an infection with the virus, so how UL144 is able to activate NF-kappa B in the presence of continued IE86 expression is unclear. Here we show that although IE86 does repress the UL144-mediated activation of a synthetic NF-kappa B promoter, it is unable to block UL144-mediated activation of the CCL22 promoter, and this lack of responsiveness to IE86 appears to be regulated by binding of the CREB transcription factor.
  • CCL27-transgenic mice show enhanced contact hypersensitivity to Th2, but not Th1 stimuli, Shinji Kagami, Hidehisa Saeki, Yuichiro Tsunemi, Koichiro Nakamura, Yoshihiro Kuwano, Mayumi Komine, Takashi Nakayama, Osamu Yoshie, Kunihiko Tamaki, EUROPEAN JOURNAL OF IMMUNOLOGY, EUROPEAN JOURNAL OF IMMUNOLOGY, 38(3), 647 - 657, Mar. 2008 , Refereed
    Summary:CCL27 is one of the CC chemokines produced by epidermal keratinocytes and is suggested to be involved in the pathogenesis of inflammatory skin diseases. To clarify the contribution of CCL27 in skin inflammation, we created transgenic C57BL/6 mice that constitutively produce CCL27 in epidermal keratinocytes. These mice had high serum CCL27 levels and did not show any phenotypical change. Thus we stimulated these mice with various reagents by single and repeated application. Interestingly, only contact hypersensitivity to repeated application with fluorescein isothiocyanate was significantly enhanced in transgenic mice compared to non-transgenic mice. Under this condition, the numbers of inflammatory cells, CCR10-positive cells, CCR4-positive cells and cutaneous lymphocyte-associated antigen-positive cells were increased, and IL-4 mRNA expression was higher in the lesional skin of transgenic mice. Increased number of mast cells and higher serum IgE levels, which were similar to atopic dermatitis, were also observed. These results indicated that CCL27 modified inflammation by attracting CCR10-positive and CCR4-positive cells into the lesional skin, and may participate in the pathogenesis of Th2-shifted skin diseases such as atopic dermatitis.
  • 1,25-Dihydroxyvitamin D-3 induces CCR10 expression in terminally differentiating human B cells, Aiko-Konno Shirakawa, Daisuke Nagakubo, Kunio Hieshima, Takashi Nakayama, Zhe Jin, Osamu Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 180(5), 2786 - 2795, Mar. 2008 , Refereed
    Summary:In the B cell lineage, CCR10 is known to be selectively expressed by plasma cells, especially those secreting IgA. In this study, we examined the regulation of CCR10 expression in terminally differentiating human B cells. As reported previously, IL-21 efficiently induced the differentiation of activated human CD19(+) B cells into IgD(-)CD38(+) plasma cells in vitro. A minor proportion of the resulting CD19(+)IgD(-)CD38(+) cells expressed CCR10 at low levels. 1,25-Dihydroxyvitamin D-3 (1,25-(OH)(2)D-3), the active metabolite of vitamine D-3, dramatically increased the proportion of CD19(+)IgD(-)CD38(+) cells expressing high levels of CCR10. The 1,25(OH)2D3 also increased the number of CCR10(+) cells expressing surface IgA, although the majority of CCR10(+) cells remained negative for surface IgA. Thus, 1,25-(OH)(2)D-3 alone may not be sufficient for the induction of IgA expression in terminally differentiating human B cells. To further determine whether 1,25-(OH)(2)D-3 directly induces CCR10 expression in terminally differentiating B cells, we next performed the analysis on the human CCR10 promoter. We identified a proximal Ets-1 site and an upstream potential vitamin D response element to be critical for the inducible expression of CCR10 by 1,25-(OH)(2)D-3. We confirmed the specific binding of Ets-1 and 1,25-(OH)(2)D-3-activated vitamin D receptor to the respective sites. In conclusion, 1,25-(OH)(2)D-3 efficiently induces CCR10 expression in terminally differentiating human B cells in vitro. Furthermore, the human CCR10 promoter is cooperatively activated by Ets-1 and vitamin D receptor in the presence of 1,25-(OH)(2)D-3.
  • Expression of CCL17 and CCL22 by latent membrane protein 1-positive tumor cells in age-related Epstein-Barr virus-associated B-cell lymphoproliferative disorder, Sumio Takegawa, Zhe Jin, Takashi Nakayama, Takashi Oyama, Kunio Hieshima, Daisuke Nagakubo, Aiko-Konno Shirakawa, Toyonori Tsuzuki, Shigeo Nakamura, Osamu Yoshie, CANCER SCIENCE, CANCER SCIENCE, 99(2), 296 - 302, Feb. 2008 , Refereed
    Summary:Age-related Epstein-Barr virus-positive (EBV+) B-cell lymphoproliferative disorder (ALPD) is a disease entity identified from a large-scale re-survey of cases diagnosed as diffuse large B-cell lymphoma. ALPD is a group of EBV+ polymorphic B-cell lymphoma typically seen in elderly patients. An age-associated decline in host immunity against EBV might be partly responsible for the pathogenesis of ALPD. Histologically, ALPD is often characterized by a minor proportion of EBV-encoded RNA-positive tumor cells in a background of extensive cellular infiltration, similar to that of classical Hodgkin's lymphoma. In contrast to Hodgkin and Reed-Sternberg cells, ALPD tumor cells are clearly positive for B cell markers CD20 and/or CD79a. Hodgkin and Reed-Sternberg cells produce various chemokines, including CCL17 and CCL22, that attract chemokine receptor CCR4-expressing Th2 cells and regulatory T cells. Previously, we have shown that EBV-immortalized B cells also produce CCL17 and CCL22 through latent membrane protein 1 (LMP1)-mediated activation of nuclear factor kappa B. Here we examined expression of CCL17 and CCL22 in ALPD. ALPD tumor cells were often heterogeneous in size in accordance with the differential expression of EBV latent genes at the single cell level. LMP1-expressing tumor cells were typically large in size and selectively positive for CCL17 and CCL22. CCR4(+) cells and forkhead box protein 3(+) regulatory T cells were abundantly present, and the majority of forkhead box protein 3(+) cells were CCR4(+). Collectively, our data show production of CCL17 and CCL22 by LMP1(+) large-sized tumor cells and accumulation of CCR4-expressing cells including regulatory T cells in ALPD.
  • Tax-inducible production of CC chemokine ligand 22 by human T cell leukemia virus type 1 (HTLV-1)-Infected T cells promotes preferential transmission of HTLV-1 to CCR4-expressing CD4(+) T cells, Kunio Hieshima, Daisuke Nagakubo, Takashi Nakayama, Aiko-Konno Shirakawa, Zhe Jin, Osamu Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 180(2), 931 - 939, Jan. 2008 , Refereed
    Summary:Adult T cell leukemia is a mature CD4(+) T cell malignancy which predominantly expresses CCR4 and is etiologically associated with human T cell leukemia virus type 1 (HTLV-1). Because HTLV-1 transmission depends on close cell-cell contacts, HTLV-1-infected T cells may preferentially interact with CCR4(+)CD4(+) T cells for efficient viral transmission. In terms of gene expression and protein secretion, we found a strong correlation between HTLV-1 Tax oncoprotein and CCL22, a CCR4 ligand, in HTLV-1-infected T cells. Transient Tax expression in an HTLV-1-negative T cell line activated the CCL22 promoter and induced CCL22. Additionally, tax gene knockdown by small interference RNA reduced CCL22 expression in the infected T cells. These findings indicate that CCL22 is a cellular target gene of Tax. In chemotaxis assays, the culture supernatants of HTLV-1-infected T cells selectively attracted CCR4(+)CD4(+) T cells in PBMCs. This was blocked by pretreating the supernatants with anti-CCL22 Ab or PBMCs with a synthetic CCR4 antagonist. In coculture experiments, primary CCR4(+)CD4(+) T cells significantly adhered to Tax-expressing cells. This adhesion was blocked by the CCR4 antagonist or pertussis toxin. Interestingly, CCR4 was redistributed to the contact region, and in some cases, this was accompanied by a polarized microtubule-organizing center, which is an indicator of virological synapse formation, in the infected T cells. Finally, anti-CCL22 Ab treatment also blocked HTLV-1 transmission to primary CD4(+) T cells in coculture experiments with HTLV-1 producer cells. Thus, HTLV-1-infected T cells produce CCL22 through Tax and selectively interact with CCR4(+)CD4(+) T cells, resulting in preferential transmission of HTLV-1 to CCR4(+)CD4(+) T cells.
  • Differing phenotypes lymphocytes in early between intraepithelial and stromal-stage tongue cancer, Fuminori Katou, Haruo Ohtani, Yoshiko Watanabe, Takashi Nakayama, Osamu Yoshie, Kenji Hashimoto, CANCER RESEARCH, CANCER RESEARCH, 67(23), 11195 - 11201, Dec. 2007 , Refereed
    Summary:The significance of tumor-infiltrating lymphocytes (TIL) has attracted much attention in relation to the prognosis of patients. We herein examined the activation status of the TILs in relation to the tumor microenvironment. By using frozen sections of human early-stage tongue cancers (n = 22), the TILs in the cancer nests and those in the cancer stroma were compared for the expression of PD-1, NKG2A, NKG2D, CD69, and Ki-67. The lymphocytes in oral lichen planus, an active immune response-mediated mucosal disease, were also analyzed for comparison purposes. All of the cancer specimens were abundantly infiltrated by CD8(+) T cells and CD56(+) natural killer (NK) cells in the stroma, as well as in the tumor nest. The tumor nest-infiltrating (intraepith,elial) CD8(+) T cells frequently expressed PD-1, an inhibitory receptor, in sharp contrast to those in the stroma or in the lichen planus. Conversely, the intraepithelial CD8(+) T cells only infrequently expressed NKG2D), an activating receptor, in contrast to those in the stroma or in the lichen planus. No intraepithelial CD8(+) T cells expressed Ki-67, a proliferation-associated marker, whereas those in the stroma frequently expressed it. Furthermore, the intraepithelial NK cells expressed NKG2A, an inhibitory receptor, more frequently than those in the stroma or the lichen planus. Collectively, the intraepithelial CD8(+) T cells and NK cells are phenotypically inactivated, whereas stromal counterparts are phenotypically just as active as those in the lichen planus. These results suggest the first-step occurrence of an immune evasion mechanism in the tumor nest of oral squamous cell carcinoma.
  • High-level expression of chemokine CXCL16 by tumor cells correlates with a good prognosis and increased tumor-infiltrating lymphocytes in colorectal cancer, Shozo Hojo, Keiichi Koizumi, Koichi Tsuneyama, Yoshihisa Arita, Zhengguo Cui, Kanna Shinohara, Takayuki Minami, Isaya Hashimoto, Takashi Nakayama, Hiroaki Sakurai, Yasuo Takano, Osamu Yoshie, Kazuhiro Tsukada, Ikuo Saiki, CANCER RESEARCH, CANCER RESEARCH, 67(10), 4725 - 4731, May 2007 , Refereed
    Summary:CXCL16 is a new member of the chemokine superfamily, which exists in a transmembrane as well as a soluble form. Its receptor CXCR6 is detected on CD4(+) T cells, CD8(+) T cells, and natural killer T cells. Here, we report a significant correlation of CXCL16 expression by tumor cells with the infiltration of T cells and prognosis in colorectal cancer (CRC). We first found that CXCL16 expression was consistently up-regulated more in tumor tissues than in normal mucosa derived from the same CRC patients. Four human CRC cell lines also expressed CXCL16 mRNA and secreted soluble CXCL16. We next examined the expression of CXCL16 and infiltration of lymphocytes in CRC specimens (n = 58) by immunohistochemistry. CRC patients with high levels of CXCL16 expression (n = 43) had higher levels of CD4(+) and CD8(+) tumor-infiltrating lymphocytes (TIL; P < 0.01) than those with tow levels of CXCL16 expression (n = 15). Furthermore, the high CXCL16 expression group showed significantly better prognosis than the low CXCL16 expression group (P < 0.05). Collectively, our data suggest that the expression of CXCL16 by tumor cells enhances the recruitment of TILs, thereby bringing about a better prognosis in CRC. Thus, CXCL16 is a new prognostic biomarker and may be useful for the development of a more effective therapeutic strategy for CRC.
  • Expression of CCR9 in HTLV-1(+) T cells and ATL cells expressing tax, Daisuke Nagakubo, Zhe Jin, Kunio Hieshima, Takashi Nakayama, Aiko-Konno Shirakawa, Yuetsu Tanaka, Hitoshi Hasegawa, Tomayoshi Hayashi, Kunihiro Tsukasaki, Yasuaki Yamada, Osamu Yoshie, INTERNATIONAL JOURNAL OF CANCER, INTERNATIONAL JOURNAL OF CANCER, 120(7), 1591 - 1597, Apr. 2007 , Refereed
    Summary:Adult T-cell leukemia (ATL) is a highly aggressive mature CD4(+) T-cell malignancy that is etiologically associated with human T-lymphotropic virus Type I (HTLV-1). ATL is characterized by frequent infiltration of lymph nodes, spleen, liver, skin and gut. Previously, we and others have shown that the majority of ATL cases are strongly positive for CCR4, which may explain the frequent skin invasion of ATL. Here, we examined whether ATL cells express CCR9, which is involved in T-cell homing to the gastrointestinal tract. Human T cell lines carrying HTLV-1 consistently expressed CCR9 together with the HTLV-1-encoded transcriptional activator Tax. Although ATL cells freshly isolated from peripheral blood hardly expressed CCR9, ATL cells cultured for I day consistently expressed CCR9 in parallel with the upregulation of Tax. Induction of Tax by Cd2+ in JPX-9, a subline of Jurkat human T cell line carrying Tax under the control of metallothionein promoter, led to upregulation of CCR9. A luciferase reporter gene under the control of the CCR9 promoter was expressed by cotransfection of an expression vector for Tax or in Cd2+-treated JPX-9 cells. Furthermore, immunohistochemical staining demonstrated that ATL cells infiltrating gastrointestinal tract were frequently positive for CCR9. Collectively, CCR9 is inducible in ATL cells expressing Tax and may play a role in the gastrointestinal involvement of ATL. (c) 2007 Wiley-Liss, Inc.
  • Survey of chemokine receptor expression reveals frequent co-expression of skin-homing CCR4 and CCR10 in adult T-cell leukemia/lymphoma, Hitomi Harasawa, Yasuaki Yamada, Kunio Hieshima, Zhe Jin, Takashi Nakayama, Osamu Yoshie, Kazuhiro Shimizu, Hiroo Hasegawa, Tomayoshi Hayashi, Yoshitaka Imaizumi, Shuichi Ikeda, Hiroshi Soda, Hisashi Soda, Sunao Atogami, Yumi Takasaki, Kunihiro Tsukasaki, Masao Tomonaga, Ken Murata, Kazuyuki Sugahara, Kazuto Tsuruda, Shimeru Kamihira, LEUKEMIA & LYMPHOMA, LEUKEMIA & LYMPHOMA, 47(10), 2163 - 2173, Oct. 2006 , Refereed
    Summary:Adult T-cell leukemia/lymphoma (ATLL) is a malignancy of mature T-cell origin with multi-organ involvement. Because the chemokine receptors play crucial roles in tissue-specific homing of mature lymphocytes, particular chemokine receptors expressed on ATLL cells may be involved in their tissue infiltration. We thus performed a comprehensive survey on the chemokine receptor expression in ATLL. ATLL cells expressed transcripts of CCR1, CCR4, CCR7, CCR8, CCR10 and CXCR4 but hardly expressed those of CCR2, CCR3, CCR5, CCR6, CCR9, CXCR1, CXCR2, CXCR3 and CXCR5. These results were confirmed at the protein level by flow cytometric analysis. Notably, patients who have skin lesions showed significantly higher levels of CCR10 mRNA expression than patients without skin lesions. ATLL cells migrated efficiently to the CCR4 ligand, CCL22, and moderately to the CCR10 ligands, CCL27 and CCL28. Moreover, ATLL skin lesions consistently contained transcripts of CCR10 and its ligands CCL27 and CCL28 besides those of CCR4 and its ligands CCL17 and CCL22 that have been reported previously. Collectively, the frequent co-expression of CCR4 and CCR10, the known pair of skin-homing chemokine receptors, may play an important role in ATLL invasion into the skin.
  • CCL17 transgenic mice show an enhanced Th2-type response to both allergic and non-allergic stimuli, Yuichiro Tsunemi, Hidehisa Saeki, Koichiro Nakamura, Daisuke Nagakubo, Takashi Nakayama, Osamu Yoshie, Shinji Kagami, Kiyo Shimazu, Takafumi Kadono, Makoto Sugaya, Mayumi Komine, Koji Matsushima, Kunihiko Tamaki, EUROPEAN JOURNAL OF IMMUNOLOGY, EUROPEAN JOURNAL OF IMMUNOLOGY, 36(8), 2116 - 2127, Aug. 2006 , Refereed
    Summary:CC chemokine ligand (CCL)17 is implicated in the pathogenesis of atopic dermatitis (AD). To study the effect of CCL17 produced by keratinocytes (KC) during inflammation, we created transgenic (Tg) mice in which CCL17 is overexpressed in KC. Th2-type contact hypersensitivity (CHS) was enhanced and Th1-type CHS was suppressed in these mice. Increased numbers of CC chemokine receptor (CCR)4(+) cells and mast cells infiltrated in Tg mice. Levels of IL-4 mRNA were higher and those of IFN-gamma mRNA were lower in both acute and chronic CHS. Higher levels of serum IgE were observed after CHS. Numbers of CCR4(+) cells among PBMC were increased in Tg mice challenged acutely on the trunk. Chronic irritation with croton oil induced dermatitis and an elevation of serum IgE levels. Tg mice showed enhanced ear swelling after tape stripping. CCL17 was thought to modify the inflammation caused by sensitizing reagents as well as irritant reagents by attracting CCR4(+) cells into the lesional skin and creating a Th2-dominant condition. AD-like conditions such as increased number of mast cells and elevated levels of serum IgE were observed. Thus, CCL17 may participate in the pathogenesis of skin diseases such as AD by regulating both allergic and irritant inflammation.
  • Novel antiviral activity of chemokines, T Nakayama, J Shirane, K Hieshima, M Shibano, M Watanabe, Z Jin, D Nagakubo, T Salto, Y Shimomura, O Yoshie, VIROLOGY, VIROLOGY, 350(2), 484 - 492, Jul. 2006 , Refereed
    Summary:Antimicrobial peptides are a diverse family of small, mostly cationic polypeptides that kill bacteria, fungi and even some enveloped viruses, while chemokines are a group of mostly cationic small proteins that induce directed migration of leukocytes through interactions with a group of seven transmembrane G protein-coupled receptors. Recent studies have shown that antimicrobial peptides and chemokines have substantially overlapping functions. Thus, while some antimicrobial peptides are chemotactic for leukocytes, some chemokines can kill a wide range of bacteria and fungi. Here, we examined a possible direct antiviral activity of chemokines against an enveloped virus HSV-1. Among 22 human chemokines examined, chemokines such as MIP-1 alpha/CCL3,MIP-1 beta/CCL4 and RANTES/CCL5 showed a significant direct antiviral activity against HSV-1. It is intriguing that these chemokines are mostly known to be highly expressed by effector CD8(+) T cells. The chemokines with a significant anti-HSV-1 activity commonly bound to HSV-1 virions via envelope glycoprotein gB. Electron microscopy revealed that the chemokines with a significant anti-HSV-1 activity were commonly capable of generating pores in the envelope of HSV-1. Thus, some chemokines have a significant direct antiviral activity against HSV-1 in vitro and may have a potential role in host defense against HSV-1 as a direct antiviral agent. (c) 2006 Elsevier Inc. All rights reserved.
  • Antagonist of interferon-inducible protein 10/CXCL10 ameliorates the progression of autoimmune sialadenitis in MRL/lpr mice, H Hasegawa, A Inoue, M Kohno, M Muraoka, T Miyazaki, M Terada, T Nakayama, O Yoshie, M Nose, M Yasukawa, ARTHRITIS AND RHEUMATISM, ARTHRITIS AND RHEUMATISM, 54(4), 1174 - 1183, Apr. 2006 , Refereed
    Summary:Objective. Mononuclear cell infiltration of the salivary glands is a major feature of Sjogren's syndrome (SS) and its animal model. Local generation of chemokines and the presence of chemokine receptors on the infiltrating cells may be involved in this process. We undertook the present study to investigate the expression of chemokines during the development of autoimmune sialadenitis in MRL/lpr mice and the therapeutic effect of chemokine antagonists on sialadenitis. Methods. NH2-terminal-truncated interferon-inducible protein 10 (IP-10)/CXCL10 analogs were transfected into a nonmetastatic fibroblastoid cell line, MRL/N-1, and injected subcutaneously into MRL/lpr mice, and the effects on sialadenitis were monitored. Results. IP-10 analogs truncated by 5 or more amino acid residues from the N-terminal failed to induce chemotaxis and calcium influx by CXCR3-expressing cells. Of these, the most potent antagonist (AT) (IP-10-AT) was a molecule with methionine added after removal of the 5 N-terminal amino acid residues. Significantly increased expression of the Th1-associated chemokines IP-10, monokine induced by interferon-gamma/ CXCL9, and interferon-inducible T cell chemo-attractant/CXCL11 was induced in the ductal epithelium by interferon-gamma produced in the salivary glands, whereas expression of the Th2-associated chemokines thymus and activation-regulated chemokine (TARC)/ CCL17 and monocyte-derived chemokine/CCL22 was almost undetectable during sialadenitis. Inoculation of IP-10-AT into MRL/lpr mice during the early stage of sialadenitis significantly reduced periductal mononuclear cell infiltration and parenchymal destruction compared with these features in control and TARC-AT-bearing mice. This was due to a significant reduction in infiltration of CXCR3+ T cells, predominantly Th1 cells, resulting in decreased interferon-gamma production. Conclusion. We prepared a novel potent IP-10 antagonist and demonstrated its ability to ameliorate the progression of autoimmune sialadenitis. This agent may provide a new therapeutic approach to SS.
  • RANKL-induced CCL22/macrophage-derived chemokine produced from osteoclasts potentially promotes the bone metastasis of lung cancer expressing its receptor CCR4, Eliane Shizuka Nakamura, Keiichi Koizumi, Mitsuo Kobayashi, Yurika Saitoh, Yoshihisa Arita, Takashi Nakayama, Hiroaki Sakurai, Osamu Yoshie, Ikuo Saiki, CLINICAL & EXPERIMENTAL METASTASIS, CLINICAL & EXPERIMENTAL METASTASIS, 23(1), 9 - 18, Mar. 2006 , Refereed
    Summary:Chemokines are now known to play an important role in cancer growth and metastasis. Here we report that differentiating osteoclasts constitutively produce CCL22 (also called macrophage-derived chemokine) and potentially promote bone metastasis of lung cancer expressing its receptor CCR4. We first examined expression of chemokines by differentiating osteoclasts. CCL22 was selectively upregulated in osteoclast-like cells derived from RAW264.7 cells and mouse bone marrow cells upon stimulation with RANKL (receptor activator of nuclear factor-kappa B ligand). In addition, a human lung cancer cell line SBC-5 that efficiently metastasized to bone when intravenously injected into NK cell-depleted SCID mice was found to express CCR4. Stimulation of SBC-5 cells with CCL22 induced cell migration and also enhanced phosphorylation of protein kinase B/Akt and extracellular signal-regulated kinase (ERK). Furthermore, immunohistochemical analysis of bone metastasis lesions demonstrated close co-localization of tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts expressing CCL22 and SBC-5 cells expressing CCR4. Collectively, these results suggest that osteoclasts may promote bone metastasis of cancer cells expressing CCR4 in the bone marrow by producing its ligand CCL22.
  • Role of the CXCL12/CXCR4 axis in peritoneal carcinomatosis of gastric cancer, K Yasumoto, K Koizumi, A Kawashima, Y Saitoh, Y Arita, K Shinohara, T Minami, T Nakayama, H Sakurai, Y Takahashi, O Yoshie, Saiki, I, CANCER RESEARCH, CANCER RESEARCH, 66(4), 2181 - 2187, Feb. 2006 , Refereed
    Summary:Peritoneal carcinomatosis is a frequent cause of death in patients with advanced gastric carcinoma. Because chemokines are now considered to play an important role in the metastasis of various malignancies, we hypothesized that they may be involved in the development of peritoneal carcinomatosis by gastric carcinoma. Human gastric carcinoma cell lines, which were all highly efficient in generating malignant ascites in nude mice upon i.p. inoculation, selectively expressed CXCR4 mRNA and protein. In particular, NUGC4 cells expressed CXCR4 mRNA at high levels and showed vigorous migratory responses to its ligand CXCL12. CXCL12 enhanced proliferation and rapid increases in phosphorylation of protein kinase B/Akt and extracellular signal-regulated kinase of NUGC4 cells. We also showed that AMD3100 (a specific CXCR4 antagonist) effectively reduced tumor growth and ascitic fluid formation in nude mice inoculated with NUGC4 cells. Additionally, we examined human clinical samples. Malignant ascitic fluids from patients with peritoneal carcinomatosis contained high concentrations of CXCL12 (4.67 ng/mL). Moreover, immunohistochemical analysis showed that 22 of 33 primary gastric tumors with peritoneal metastasis were positive for CXCR4 expression (67%), whereas only 4 of 16,with other distant metastasis were positive (25%). Notably, 22 of 26 CXCR4-expressing primary tumors developed peritoneal metastases (85%). CXCR4 positivity of primary gastric carcinomas significantly correlated with the development of peritoneal carcinomatosis (P < 0.001). Collectively, our results strongly suggest that the CXCR4/CXC12 axis plays an important role in the development of peritoneal carcinomatosis from gastric carcinoma. Thus, CXCR4 may be a potential therapeutic target for peritoneal carcinomatosis of gastric carcinoma.
  • Dopamine selectively induces migration and homing of naive CD8(+) T cells via dopamine receptor D3, Y Watanabe, T Nakayama, D Nagakubo, K Hieshima, Z Jin, F Katou, K Hashimoto, O Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 176(2), 848 - 856, Jan. 2006 , Refereed
    Summary:The nervous systems affect immune functions by releasing neurohormones and neurotransmitters. A neurotransmitter dopamine signals via five different seven-transmembrane G protein-coupled receptors termed D1 to D5. The secondary lymphoid tissues are highly innervated by sympathetic nerve fibers that store dopamine at high contents. Lymphocytes also produce dopamine. In this study, we examined expression and function of dopamine receptors in lymphocytes. We found that D3 was the predominant subtype of dopamine receptors in the secondary lymphoid tissues and selectively expressed by naive CD8(+) T cells of both humans and mice. Dopamine induced calcium flux and chemotaxis in mouse L1.2 cells stably expressing human D3. These responses were almost completely inhibited by pertussis toxin, indicating that D3 was coupled with the G alpha i class of G proteins. Consistently, dopamine selectively induced chemotactic responses in naive CD8(+) T cells of both humans and mice in a manner sensitive to pertussis toxin and D3 antagonists. Dopamine was highly synergistic with CCL19, CCL21, and CXCL12 in induction of chemotaxis in naive CD8(+) T cells. Dopamine selectively induced adhesion of naive CD8(+) T cells to fibronectin and ICAM-1 through activation of integrins. Intraperitoneal injection of mice with dopamine selectively attracted naive CD8(+) T cells into the peritoneal cavity. Treatment of mice with a D3 antagonist U-99194A selectively reduced homing of naive CD8(+) T cells into lymph nodes. Collectively, naive CD8(+) T cells selectively express D3 in both humans and mice, and dopamine plays a significant role in migration and homing of naive CD8(+) T cells via D3.
  • Selective infiltration of CCR5(+)CXCR3(+) T lymphocytes in human colorectal carcinoma, H Musha, H Ohtani, T Mizoi, M Kinouchi, T Nakayama, K Shiiba, K Miyagawa, H Nagura, O Yoshie, Sasaki, I, INTERNATIONAL JOURNAL OF CANCER, INTERNATIONAL JOURNAL OF CANCER, 116(6), 949 - 956, Oct. 2005 , Refereed
    Summary:T cell infiltration in colorectal cancer is associated with a favorable prognosis, suggesting an occurrence of a certain degree of anti-tumor immunity. T helper type 1 (Th1) and Th2 cells are now known to selectively express CC-chemokine receptor 5 (CCR5)/CXC-chemokine receptor 3 (CXCR3) and CCR4, respectively. To clarify the mechanism of T cell infiltration, we examined in situ expression of these chemokine receptors and their respective chemokine ligands in 40 cases of human colorectal cancer. Immunohistochemistry showed a predominant accumulation of T cells expressing CCR5 and CXCR3 mainly along the invasive margin, whereas those expressing CCR4 were rare. Flow cytometric analysis showed that more than half of CD8(+) T cells and a fraction of CD4(+) cells isolated from fresh tumor tissues co-expressed CCR5 and CXCR3, and CD8(+) T cells and CD4+ cells predominantly produced interferon-gamma (IFN-gamma) over interleukin-4 (IL-4) after in vitro stimulation. RANTES/CCL5, a ligand of CCR5, was localized within infiltrating CD8+ T cells in a granular pattern, whereas IP-10/CXCL10, a ligand of CXCR3, was localized in cancer cells and macrophages along the invasive margin. These data were consistent with an active recruitment of T cells expressing CCR5 or CXCR3 into the invasive margin of colorectal cancer. With the previous clinicopathological studies showing a favorable prognostic impact of T cell infiltration in colorectal cancer, our study supports the occurrence of a certain level of Th1-shifted cellular immune responses in human colorectal cancer. (C) 2005 Wiley-Liss, Inc.
  • Anti-tumor responses induced by chemokine CCL19 transfected into an ovarian carcinoma model via fiber-mutant adenovirus vector, JQ Gao, T Sugita, N Kanagawa, K Iida, N Okada, H Mizuguchi, T Nakayama, T Hayakawa, O Yoshie, Y Tsutsumi, T Mayumi, S Nakagawa, BIOLOGICAL & PHARMACEUTICAL BULLETIN, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 28(6), 1066 - 1070, Jun. 2005 , Refereed
    Summary:Considerable attention has recently been paid to the application of chemokines to cancer immunotherapy because of their chemotactic affinity for a variety of immune cells and because several chemokines are strongly angiostatic. In the present study, the recombinant adenovirus vectors encoding chemokine CCL19 or XCL1 in an El cassette (AdRGD-mCCL19 and AdRGD-mXCL1) were developed. The constructed fiber-mutant adenovirus vector, which contained the integrin-targeting Arg-Gly-Asp (RGD) sequence in the fiber knob, notably enhanced the transfection efficiency to OV-HM ovarian carcinoma cells compared to that induced by conventional adenovirus vector. The results of an in vitro chemotaxis assay for chemokine-encoding vector demonstrated that both AdRGD-mCCL19 and AdRGD-mXCL1 could induce the migration of cells expressing specific chemokine receptors. Of the two chemokine-en coding vectors evaluated in vivo, AdRGD-mCCL19 showed significant tumor-suppressive activity in R6C3F1 mice via transduction into OV-HM cells, whereas XCL1 did not exhibit any notable anti-tumor effects, suggesting that CCL19 may be a candidate for cancer immunotherapy.
  • CC chemokine ligands 25 and 28 play essential roles in intestinal extravasation of IgA antibody-secreting cells, K Hieshima, Y Kawasaki, H Hanamoto, T Nakayama, D Nagakubo, A Kanamaru, O Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 173(6), 3668 - 3675, Sep. 2004 , Refereed
    Summary:CCL25 (also known as thymus-expressed chemokine) and CCL28 (also known as mucosae-associated epithelial chemokine) play important roles in mucosal immunity by recruiting IgA Ab-secreting cells (ASCs) into mucosal lamina propria. However, their exact roles in vivo still remain to be defined. In this study, we first demonstrated in mice that IgA ASCs in small intestine expressed CCR9, CCR10, and CXCR4 on the cell surface and migrated to their respective ligands CCL25, CCL28, and CXCL12 (also known as stromal cell-derived factor 1), whereas IgA ASCs in colon mainly expressed CCR10 and CXCR4 and migrated to CCL28 and CXCL12. Reciprocally, the epithelial cells of small intestine were immunologically positive for CCL25 and CCL28, whereas those of colon were positive for CCL28 and CXCL12. Furthermore, the venular endothelial cells in small intestine were positive for CCL25 and CCL28, whereas those in colon were positive for CCL28, suggesting their direct roles in extravasation of IgA ASCs. Consistently, in mice orally immunized with cholera toxin (CT), anti-CCL25 suppressed homing of CT-specific IgA ASCs into small intestine, whereas anti-CCL28 suppressed homing of CT-specific IgA ASCs into both small intestine and colon. Reciprocally, CT-specific ASCs and IgA titers in the blood were increased in mice treated with anti-CCL25 or anti-CCL28. Anti-CXCL12 had no such effects. Finally, both CCL25 and CCL28 were capable of enhancing alpha(4) integrin-dependent adhesion of IgA ASCs to mucosal addressin cell adhesion molecule-1 and VCAM-1. Collectively, CCL25 and CCL28 play essential roles in intestinal homing of IgA ASCs primarily by mediating their extravasation into intestinal lamina propria.
  • Liver-expressed chemokine/CC chemokine ligand 16 attracts eosinophils by interacting with histamine H4 receptor, T Nakayama, Y Kato, K Hieshima, D Nagakubo, Y Kunori, T Fujisawa, O Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 173(3), 2078 - 2083, Aug. 2004 , Refereed
    Summary:Liver-expressed chemokine (LEC)/CCL16 is a human CC chemokine that is constitutively expressed by the liver parenchymal cells and present in the normal plasma at high concentrations. Previous studies have shown that CCL16 is a low-affinity ligand for CCR1, CCR2, CCR5, and CCR8 and attracts monocytes and T cells. Recently, a novel histamine receptor termed type 4 (H4) has been identified and shown to be selectively expressed by eosinophils and mast cells. In this study, we demonstrated that CCL16 induced pertussis toxin-sensitive calcium mobilization and chemotaxis in murine L1.2 cells expressing H4 but not those expressing histamine receptor type 1 (H1) or type 2 (H2). CCL16 bound to H4 with a K-d of 17 nM. By RT-PCR, human and mouse eosinophils express H4 but not H3. Accordingly, CCL16 induced efficient migratory responses in human and mouse eosinophils. Furthermore, the responses of human and mouse eosinophils to CCL16 were effectively suppressed by thioperamide, an antagonist for H3 and H4. Intravenous injection of CCL16 into mice induced a rapid mobilization of eosinophils from bone marrow to peripheral blood, which was also suppressed by thioperamide. Collectively, CCL16 is a novel functional ligand for H4 and may have a role in trafficking of eosinophils.
  • Chemokines generally exhibit scavenger receptor activity through their receptor-binding domain, T Shimaoka, T Nakayama, K Hieshima, N Kume, N Fukumoto, M Minami, K Hayashida, T Kita, O Yoshie, S Yonehara, JOURNAL OF BIOLOGICAL CHEMISTRY, JOURNAL OF BIOLOGICAL CHEMISTRY, 279(26), 26807 - 26810, Jun. 2004 , Refereed
    Summary:Chemokines are a family of cytokines that induce directed migration of various types of leukocytes through specific interactions with a group of seven transmembrane receptors. Scavenger receptors are a heterogenous family of transmembrane molecules that commonly bind and uptake oxidized low density lipoprotein and bacteria. Here, we show that not only CXC chemokine 16 (CXCL16)/SR-PSOX, a transmembrane chemokine with scavenger receptor activity, but also 12 out of 15 chemokines examined efficiently bound scavenger receptor ligands in competition with cells expressing their specific chemokine receptors. Furthermore both the chemotactic and scavenger receptor activities of SR-PSOX/CXCL16 were similarly impaired in a series of mutants altered in the chemokine domain, indicating that SR-PSOX/CXCL16 binds scavenger receptor ligands as well as CXCR6 using highly overlapping binding motifs. Taken together, chemokines generally have scavenger receptor-like activity through their receptor-binding domain, suggesting a close evolutionary relationship between chemokines and scavenger receptors.
  • Corneal epithelial cells and stromal keratocytes efficently produce CC chemokine-ligand 20 (CCL20) and attract cells expressing its receptor CCR6 in mouse herpetic stromal keratitis, J Shirane, T Nakayama, D Nagakubo, D Izawa, K Hieshima, Y Shimomura, O Yoshie, CURRENT EYE RESEARCH, CURRENT EYE RESEARCH, 28(5), 297 - 306, May 2004 , Refereed
    Summary:Purpose. CC chemokine-ligand 20 (CCL20) is known to be selectively expressed by surface-lining mucosal epithelial cells and skin epidermal keratinocytes and to attract cells such as immature dendritic cells and effector T cells via CCR6. This study evaluated the ability of corneal epithelial cells and stromal keratocytes to produce CCL20 in vitro and in vivo. Methods. Human corneal epithelial cells (HCE) and corneal keratocytes (HCK) were treated without or with various cytokines and expression of CCL20 mRNA and secreion of its protein were evaluated by RT-PCR and ELISA. Induction of CCL20 mRNA in HCE and HCK was also examined upon in vitro infection with HSV-1. Using a mouse model of herpetic stromal keratitis (HSK), induction of CCL20 expression and accumulation of cells expressing CCR6 were evaluated by RT-PCR and immunohistochemistry. Results. Not only corneal epithelial cells but also stromal keratocytes efficiently expressed CCL20 mRNA and protein upon stimulation with IL-1beta and TNF-alpha. In vitro infection with HSV-1 also induced CCL20 mRNA in both types of cells. In a mouse herpetic stromal keratitis model, prominent accumulation of CCL20 and CCR6 mRNA was revealed in HSV-1-infected corneas. Furthermore, immunohistochemistry demonstrated production of CCL20 by corneal epithelial cells as well as, stromal keratocytes and stromal infiltration of DEC205(+) dendritic cells, CD4(+) T cells and CD8(+) T cells. Double staining revealed that CCR6-expressing cells were mostly MHC class II+ dendritic cells. Conclusions. Not only epithelial cells but also stromal keratocytes are efficient producers of CCL20 in the cornea and recruit CCR6-expressing cells such as dendritic cells into inflamed cornea.
  • Anti-tumor activity of chemokine is affected by both kinds of tumors and the activation state of the host's immune system: implications for chemokine-based cancer immunotherapy, N Okada, JQ Gao, A Sasaki, M Niwa, Y Okada, T Nakayama, O Yoshie, H Mizuguchi, T Hayakawa, T Fujita, A Yamamoto, Y Tsutsumi, T Mayumi, S Nakagawa, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 317(1), 68 - 76, Apr. 2004 , Refereed
    Summary:In this study, we screened the anti-tumor activity of murine chemokines including CCL17. CCL19, CCL20, CCL21, CCL22, CCL27, XCL1, and CX3CL1 by inoculating murine B16BL6, CT26, or OV-HM tumor cells, all of which were transfected with chemokine-expressing fiber-mutant adenovirus vector, into immunocompetent mice. A tumor-suppressive effect was observed in mice inoculated with CCL19/B16BL6 and XCK1/B16BL6, and CCL22/OV-HM showed considerable retardation in tumor growth. In the cured mice inoculated with CCL22/OV-HM, a long-term specific immune protection against parental tumor was developed. However, we were unable to identify the chemokine that had a suppressive activity common to all three tumor models. Furthermore, an experiment using chemokine-transfected B16BL6 cells was also performed on mice sensitized with melanoma-associated antigen. A drastic enhancement of the frequency of complete rejection was observed in mice inoculated with CCL17-, CCL19-, CCL22-, and CCL27-transfected B16BL6. Altogether, our results suggest that the tumor-suppressive activity of chemokine-gene immunotherapy is greatly influenced by the kind of tumor and the activation state of the host's immune system. (C) 2004 Elsevier Inc. All rights reserved.
  • Infiltration of CD8(+) T cells containing RANTES/CCL5(+) cytoplasmic granules in actively inflammatory lesions of human chronic gastritis, N Ohtani, H Ohtani, T Nakayama, H Naganuma, E Sato, T Imai, H Nagura, O Yoshie, LABORATORY INVESTIGATION, LABORATORY INVESTIGATION, 84(3), 368 - 375, Mar. 2004 , Refereed
    Summary:Chronic gastritis is frequently associated with infection of Helicobacter pylori and characterized by tissue infiltration of neutrophils, lymphocytes, and plasma cells. To address the mechanism of lymphocyte infiltration in chronic gastritis, we examined the expression of chemokines and their receptors using frozen sections of chronic gastritis, obtained from 23 patients who underwent gastrectomy for gastric cancer. By immuno histochemistry, lymphocytes in inflamed gastric mucosa expressed CCR5 abundantly, CXCR3 less frequently, and CCR4 sparsely. The numbers of CCR5(+) cells, which were composed of mainly CD8(+) and partly CD4(+) T cells, were positively correlated with the degree of neutrophil infiltration, and decreased in areas with intestinal metaplasia or mucosal atrophy. RANTES/CCL5, one of the ligands of CCR5, was localized mainly in CD8(+) and partly CD4(+) T cells with a characteristic dotted pattern, and such lymphocytes were most densely distributed around the neck region of gastric glands. In situ hybridization confirmed the expression of CCLS mRNA in these cells, and immunoelectron microscopy revealed localization of CCL5 in the membrane-bound granules, which most probably corresponded to the cytolytic granules of cytotoxic T cells. The numbers of CCL5(+) lymphocytes showed a close correlation with the degree of neutrophil infiltration and markedly decreased in intestinal metaplasia. In conclusion, our data suggest that, together with neutrophils, CCL5+ T cells, presumably activated cytotoxic T cells, would play important roles in the active inflammatory process of chronic gastritis. Our data also suggest a self-recruiting mechanism involving CCR5 and CCL5 for tissue accumulation of such T cells.
  • Expression of CCL28 by Reed-Sternberg cells defines a major subtype of classical Hodgkin's disease with frequent infiltration of eosinophils and/or plasma cells, H Hanamoto, T Nakayama, H Miyazato, S Takegawa, K Hieshima, Y Tatsumi, A Kanamaru, O Yoshie, AMERICAN JOURNAL OF PATHOLOGY, AMERICAN JOURNAL OF PATHOLOGY, 164(3), 997 - 1006, Mar. 2004 , Refereed
    Summary:Classical Hodgkin's disease (HD) is characterized by rare neoplastic Hodgkin and Reed-Sternberg (H-RS) cells within abundant reactive cellular backgrounds. In most cases, H-RS cells originate from the B-cell lineage, but their immunophenotypes are unusual. Here we newly found frequent expression of chemokine receptors CXCR6 and CCR10 and their respective ligands CXCL16 and CCL28 in RD-derived cell lines. CCR10 is known to be selectively expressed by plasma cells, whereas CCL28 attracts eosinophils via CCR3 and plasma cells via CCR10 and CCR3. Therefore, we examined their expression in HD tissues by immunohistochemistry. We found that H-RS cells in 15 of 19 cases were positive for CCL28. Among them, seven cases were also positive for CCR10, suggesting a potential autocrine effect. In situ hybridization confirmed the expression of CCL28 mRNA in H-RS cells. The CCL28 positivity in H-RS cells did not significantly correlate with that of LMP-1, CCL17, CCL22, or CCL11. However, it significantly correlated with the background accumulation of eosinophils, plasma cells, and CCR10(+) cells. Thus, the production of CCL28 by H-RS cells may play a major role in tissue accumulation of eosinophils and/or plasma cells in classical HD. The frequent expression of CCR10 in H-RS cells themselves also supports their close relationship to plasma cells.
  • Cell surface-anchored SR-PSOX/CXC chemokine ligand 16 mediates firm adhesion of CXC chemokine receptor 6-expressing cells, T Shimaoka, T Nakayama, N Fukumoto, N Kume, S Takahashi, J Yamaguchi, M Minami, K Hayashida, T Kita, J Ohsumi, O Yoshie, S Yonehara, JOURNAL OF LEUKOCYTE BIOLOGY, JOURNAL OF LEUKOCYTE BIOLOGY, 75(2), 267 - 274, Feb. 2004 , Refereed
    Summary:Direct contacts between dendritic cells (DCs) and T cells or natural killer T (NKT) cells play important roles in primary and secondary immune responses. SR-PSOX/CXC chemokine ligand 16 (CXCL16), which is selectively expressed on DCs and macrophages, is a scavenger receptor for oxidized low-density lipoprotein and also the chemokine ligand for a G protein-coupled receptor CXC chemokine receptor 6 (CXCR6), expressed on activated T cells and NKT cells. SR-PSOX/CXCL16 is the second transmembrane-type chemokine with a chemokine domain fused to a mucin-like stalk, a structure very similar to that of fractalkine (FNK). Here, we demonstrate that SR-PSOX/CXCL16 functions as a cell adhesion molecule for cells expressing CXCR6 in the same manner that FNK functions as a cell adhesion molecule for cells expressing CX3C chemokine receptor 1 (CX(3)CR1) without requiring CX(3)CR1-mediated signal transduction or integrin activation. The chemokine domain of SR-PSOX/CXCL16 mediated the adhesion of CXCR6-expressing cells, which was not impaired by treatment with pertussis toxin, a Galphai protein blocker, which inhibited chemotaxis of CXCR6-expressing cells induced by SR-PSOX/CXCL16. Furthermore, the adhesion activity was up-regulated by treatment of SR-PSOX/CXCL16-expressing cells with a metalloprotease inhibitor, which increased surface expression levels of SRPSOX/CXCL16. Thus, SR-PSOX/CXCL16 is a unique molecule that not only attracts T cells and NKT cells toward DCs but also supports their firm adhesion to DCs.
  • Selective induction of Th2-attracting chemokines CCL17 and CCL22 in human B cells by latent membrane protein 1 of Epstein-Barr virus, T Nakayama, K Hieshima, D Nagakubo, E Sato, M Nakayama, K Kawa, O Yoshie, JOURNAL OF VIROLOGY, JOURNAL OF VIROLOGY, 78(4), 1665 - 1674, Feb. 2004 , Refereed
    Summary:Chemokines are likely to play important roles in the pathophysiology of diseases associated with Epstein-Barr virus (EBV). Here, we have analyzed the repertoire of chemokines expressed by EBV-infected B cells. EBV infection of B cells induced expression of TARC/CCL17 and MDC/CCL22, which are known to attract Th2 cells and regulatory T cells via CCR4, and also upregulated constitutive expression of MIP-1alpha/CCL3, MIP-1beta/CCL4, and RANTES/CCL5, which are known to attract Th1 cells and cytotoxic T cells via CCR5. Accordingly, EBV-immortalized B cells secreted these chemokines, especially CCL3, CCL4, and CCL22, in large quantities. EBV infection or stable expression of LMP1 also induced CCL17 and CCL22 in a B-cell line, BJAB. The inhibitors of the TRAF/NF-kappaB pathway (BAY11-7082) and the p38/ATF2 pathway (SB202190) selectively suppressed the expression of CCL17 and CCL22 in EBV-immortalized B cells and BJAB-LMP1. Consistently, transient-transfection assays using CCL22 promoter-reporter constructs demonstrated that two NF-kappaB sites and a single AP-1 site were involved in the activation of the CCL22 promoter by LMP1. Finally, serum CCL22 levels were significantly elevated in infectious mononucleosis. Collectively, LMP1 induces CCL17 and CCL22 in EBV-infected B cells via activation of NF-kappaB and probably ATF2. Production of CCL17 and CCL22, which attract Th2 and regulatory T cells, may help EBV-infected B cells evade immune surveillance by Th1 cells. However, the concomitant production of CCL3, CCL4, and CCL5 by EBV-infected B cells may eventually attract Th1 cells and cytotoxic T cells, leading to elimination of EBV-infected B cells at latency III and to selection of those with limited expression of latent genes.
  • Cutting edge: SR-PSOX/CXC chemokine ligand 16 mediates bacterial phagocytosis by APCs through its chemokine domain, T Shimaoka, T Nakayama, N Kume, S Takahashi, J Yamaguchi, M Minami, K Hayashida, T Kita, J Ohsumi, O Yoshie, S Yonehara, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 171(4), 1647 - 1651, Aug. 2003 , Refereed
    Summary:SR-PSOX and CXC chemokine ligand (CXCL) 16, which were originally identified as a scavenger receptor and a transmembrane-type chemokine, respectively, are indicated to be identical, In this study, we demonstrate that membrane-bound SR-PSOX/CXCL16 mediates adhesion and phagocytosis of both Gram-negative and Gram-positive bacteria. Importantly, our prepared anti-SR-PSOX mAb, which suppressed chemotactic activity of SR-PSOX, signficantly inhibited bacterial phagocytosis by human APCs including dendritic cells. Various scavenger receptor ligands inhibited the bacterial phagocytosis of SR-PSOX. In addition, the recognition specificity for bacteria was determined by only the chemokine domain of SR-PSOX/CXCL16. Thus, SR-PSOXICXCL16 may play an important role in facilitating uptake of various pathogens and chemotaxis of T and NKT cells by APCs through its chemokine domain.
  • Antitumor effect by interleukin-11 receptor alpha-locus chemokine/CCL27, introduced into tumor cells through a recombinant adenovirus vector, JQ Gao, Y Tsuda, K Katayama, T Nakayama, Y Hatanaka, Y Tani, H Mizuguchi, T Hayakawa, O Yoshie, Y Tsutsumi, T Mayumi, S Nakagawa, CANCER RESEARCH, CANCER RESEARCH, 63(15), 4420 - 4425, Aug. 2003 , Refereed
    Summary:In this study, we examined. antitumor activity of a mouse CC chemokine ILC/CCL27 and a mouse CX3C chemokine fractalkine/CX(3)CL1 in vivo. We generated recombinant adenovirus vectors with a fiber mutation, encoding mILC (Ad-RGD-mILC) and mFKN (Ad-RGD-mFKN). We confirmed tumor cells infected with Ad-RGD-mILC and Ad-RGD-mFKN to express and release these chemokines. Tumor rejection experiments in vivo were carried out by inoculating OV-HM cells infected with Ad-RGD-mILC or Ad-RGD-mFKN into immunocompetent mice. mILC significantly suppressed the tumor growth, whereas no such significant effect was observed by mFKN. The antitumor activity induced by mILC was T cell dependent, involving both CD4(+) and CD8(+) T cells. Immunohistochemical analysis revealed accumulation of both CD3(+) lymphocytes and NK cells in the tumor tissue transduced with mILC and mFKN. However, there was a significant difference in the distribution of infiltrating cells. Furthermore, mFKN appeared to hake an angiogenic activity, which might have masked its tumor suppressive activity. Collectively, ILC/CCL27 may be a good candidate molecule for cancer gene therapy.
  • Infiltrating CD8(+) T cells in oral lichen planus predominantly express CCR5 and CXCR3 and carry respective chemokine ligands RANTES/CCL5 and IP-10/CXCL10 in their cytolytic granules - A potential self-recruiting mechanism, W Iijima, H Ohtani, T Nakayama, Y Sugawara, E Sato, H Nagura, O Yoshie, T Sasano, AMERICAN JOURNAL OF PATHOLOGY, AMERICAN JOURNAL OF PATHOLOGY, 163(1), 261 - 268, Jul. 2003 , Refereed
    Summary:Lichen planus is a chronic inflammatory disease of the skin and oral mucosa in which the cell-mediated cytotoxicity is regarded as a major mechanism of pathogenesis. To understand its pathophysiology further, the present study examined the in situ expression of chemokines and chemokine receptors in oral lichen planus. Immunohistochemical. analysis of 15 cases has consistently revealed that infiltrating CD4(+) and CD8(+) T cells in the submucosa predominantly expressed CCR5 and CXCR3. Furthermore, infiltrating T cells, particularly CD8(+) T cells, were positive for RANTES/CCL5 and IP-10/CXCL10, the ligands of CCR5 and CXCR3, respectively. By immunoelectron microscopy, these chemokines were localized in the cytolytic granules of CD8(+) T cells. Lesional keratinocytes also overexpressed the ligands of CXCR3, namely, MIG/CXCL9, CXCL10, and I-TAC/CXCL11. Our data suggest that the chemokines signaling via CCR5 and CXCR3, which are known to be selectively expressed by type 1 T cells, are predominantly involved in T-cell infiltration of oral lichen planus. Furthermore, the presence of CCL5 and CXCL10 in the cytolytic granules of tissue-infiltrating CD8(+) T cells expressing CCR5 and CXCR3 reveals a potential self-recruiting mechanism involving activated effector cytotoxic T cells.
  • Cutting edge: Profile of chemokine receptor expression on human plasma cells accounts for their efficient recruitment to target tissues, T Nakayama, K Hieshima, D Izawa, Y Tatsumi, A Kanamaru, O Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 170(3), 1136 - 1140, Feb. 2003 , Refereed
    Summary:We systematically examined the repertoire of chemokine receptors expressed by human plasma cells. Fresh bone marrow plasma cells and myeloma cells consistently expressed CXCR4, CXCR6, CCR10, and CCR3. Accordingly, plasma cells responded to. their respective ligands in chemotaxis and very late Ag-4-dependent cell adhesion to fibronectin. Immobilized CXC chemokine ligand (CXCL)16, a novel transmembrane-type chemokine and CXCR6 ligand, also directly induced adhesion of plasma cells without requiring G(alphai) signaling or divalent cations. Furthermore, we revealed consistent expression of CXCL12 (CXCR4 ligand), CXCL16 (CXCR6 ligand), and CC chemokine ligand 28 (CCR10 and CCR3 ligand) in tissues enriched with plasma cells including bone marrow, and constitutive expression. of CXCL12, CXCL16, and CC chemokine ligand 28 by cultured human bone marrow stromal cells. Collectively, plasma cells are likely to be recruited to bone marrow and other target tissues via CXCR4, CXCR6, CCR10, and CCR3. CXCR6 may also contribute to tissue localization of plasma cells through its direct binding to membrane-anchored CXCL16.
  • CCL28 has dual roles in mucosal immunity as a chemokine with broad-spectrum antimicrobial activity, K Hieshima, H Ohtani, M Shibano, D Izawa, T Nakayama, Y Kawasaki, F Shiba, M Shiota, F Katou, T Saito, O Yoshie, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 170(3), 1452 - 1461, Feb. 2003 , Refereed
    Summary:CCL28 is a CC chemokine signaling via CCR10 and CCR3 that is selectively expressed in certain mucosal tissues such as exocrine glands, trachea, and colon. Notably, these tissues commonly secrete low-salt fluids. RT-PCR analysis demonstrated that salivary glands expressed CCL28 mRNA at the highest levels among various mouse tissues. Single cells prepared from mouse parotid glands indeed contained a major fraction of CD3(-)B220(low) cells that expressed CCR10 at high levels and CCR3 at low levels and responded to CCL28 in chemotaxis assays. Morphologically, these cells are typical plasma cells. By immunohistochemistry, acinar epithelial cells in human and mouse salivary glands were strongly positive for CCL28. Furthermore, human saliva and milk were found to contain CCL28 at high concentrations. Moreover, the C terminus of human CCL28 has a significant sequence similarity to histatin-5, a histidine-rich candidacidal peptide in human saliva. Subsequently, we demonstrated that human and mouse CCL28 had a potent antimicrobial activity against Candida albicans, Gram-negative bacteria, and Gram-positive bacteria. The C-terminal 28-aa peptide of human CCL28 also displayed a selective candidacidal activity. In contrast, CCL27, which is most similar to CCL28 and shares CCR10, showed no such potent antimicrobial activity. Like most other antimicrobial peptides, CCL28 exerted its antimicrobial activity in low-salt conditions and rapidly induced membrane permeability in target microbes. Collectively, CCL28 may play dual roles in mucosal immunity as a chemoattractant for cells expressing CCR10 and/or CCR3 such as plasma cells and also as a broad-spectrum antimicrobial protein secreted into low-salt body fluids.
  • Differential expression of CCL19 by DC-Lamp(+) mature dendritic cells in human lymph node versus chronically inflamed skin, F Katou, H Ohtani, T Nakayama, H Nagura, O Yoshie, K Motegi, JOURNAL OF PATHOLOGY, JOURNAL OF PATHOLOGY, 199(1), 98 - 106, Jan. 2003 , Refereed
    Summary:De novo formation of lymphoid tissue is one of the characteristic features of chronic inflammation. The formation of T cell-mature dendritic cell (DC) clusters has been previously demonstrated in chronically inflamed skin infected with Candida albicans. A functional similarity was also found between chronic inflammation and the T-cell zone of lymph nodes (LNs), since a substantial fraction of phenotypically mature DCs in both tissues expressed CCL22 (macrophage-derived chemokine; MDC) and were closely surrounded by memory-type T cells expressing its receptor, CCR4. To analyse the nature of T cell-mature DC interactions further in chronically inflamed skin and LNs, the present study focuses on another chemokine system, namely CCL19 (EBI1 ligand chemokine; ELC), CCL21 (secondary lymphoid tissue chemokine; SLC) and their shared receptor, CCR7. RT-PCR analysis revealed expression of CCL19, CCL21, and CCR7 at high levels in LNs and at low levels in inflamed skin. Using immunohistochemistry, the majority of DC-Lamp(+) mature DCs in the T-cell area of LNs expressed CCL19 and were surrounded by CCR7(+) naive-type lymphocytes, while CCL21 was expressed in reticular stromal cells and vascular endothelial cells. Very few mature DCs in LNs were found to express CCR7. In contrast, the majority of DC-Lamp+ mature DCs in inflamed skin were totally negative for CCL19 and were surrounded by CCR7(-) memory-type T cells. Furthermore, CCL21 expression in the inflamed skin was detected in dermal lymphatic endothelial cells and rare CCR7(+) mature DCs were mostly seen within the lymphatic vessels. In normal skin, on the other hand, no cells immunoreactive for CCL19, CCL21, or CCR7 were found. The present study thus reveals a striking difference in the function of mature DCs between LNs and chronically inflamed skin. Copyright (C) 2002 John Wiley Sons, Ltd.
  • IFN-gamma-inducible expression of thymus and activation-regulated chemokine/CCL17 and macrophage-derived chemokine/CCL22 in epidermal keratinocytes and their roles in atopic dermatitis, T Horikawa, T Nakayama, Hikita, I, H Yamada, R Fujisawa, T Bito, S Harada, A Fukunaga, D Chantry, PW Gray, A Morita, R Suzuki, T Tezuka, M Ichihashi, O Yoshie, INTERNATIONAL IMMUNOLOGY, INTERNATIONAL IMMUNOLOGY, 14(7), 767 - 773, Jul. 2002 , Refereed
    Summary:Thymus and activation-regulated chemokine (TARC)/CCL17 and macrophage-derived chemokine (MDC)/CCL22 are a pair of CC chemokines known to selectively attract T(h)2 type memory T cells via CCR4. Here we examined circulating levels of TARC and MDC in patients with atopic dermatitis (AD) and control subjects by using plasma samples, which reflect blood contents of chemokines more accurately than serum samples. The plasma levels of TARC and MDC were significantly elevated in AD patients. These values also strongly correlated with disease severity and serum lactate dehydrogenase levels, and weakly correlated with serum total IgE levels and blood eosinophilia. Previous studies demonstrated TARC immunoreactivity in the epidermal layer of AD lesional skin and production of TARC by a human keratinocytic cell line HaCaT upon stimulation with IFN-gamma. Here we demonstrated MDC immunoreactivity in the epidermal layer of AD skin at levels stronger than that of TARC. Furthermore, primary epidermal keratinocytes expressed both TARC and MDC mRNA upon stimulation with IFN-gamma, but efficiently secreted only MDC. These results suggest a post-transcriptional regulation in TARC production. IFN-gamma also induced TARC and MDC mRNA in mouse skin. Collectively, both TARC and MDC play important roles in the local accumulation of T(h)2 cells in AD lesional skin. Production of T(h)2-attracting chemokines by epidermal keratinocytes upon treatment with IFN-gamma, which is also the potent inducer of T(h)1-attracting chemokines, may underline the pivotal role of IFN-gamma in the chronic phase of AD where both T(h)1 and T(h)2 responses are mixed.
  • Presence of high contents of thymus and activation-regulated chemokine in platelets and elevated plasma levels of thymus and activation-regulated chemokine and macrophage-derived chemokine in patients with atopic dermatitis, T Fujisawa, R Fujisawa, Y Kato, T Nakayama, A Morita, H Katsumata, H Nishimori, K Iguchi, H Kamiya, PW Gray, D Chantry, R Suzuki, O Yoshie, JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 110(1), 139 - 146, Jul. 2002 , Refereed
    Summary:Background: T(H)2 cells and eosinophils selectively express CC chemokine receptor 4 and CCR3, respectively, and their chemokine ligands are likely to play important roles in the pathogenesis of atopic dermatitis (AD). Objective: The purpose of this study was to demonstrate the presence of thymus and activation-regulated chemokine (TARC) in platelets and its release during clotting and to evaluate the circulating levels of TARC, macrophage-derived chemokine (MDC), and eotaxin in control subjects and patients with AD. Methods: We compared plasma and serum contents of TARC, MDC, and eotaxin. We measured TARC contents in platelet lysates. We analyzed the correlation of plasma levels of TARC, MDC, and eotaxin with various clinicolaboratory parameters in patients with AD. Results: Serum contents of TARC rapidly, increased during clotting, whereas those or MDC and eotaxin increased only slightly. We demonstrated that platelets contained TARC, and its levels were dramatically elevated in patients with AD. Platelets also released TARC on stimulation with thrombin. We therefore evaluated circulating levels of these chemokines in control subjects and patients with AD by using plasma samples. Plasma TARC levels were significantly increased in patients with AD (P <.0001) and showed significant correlations with severity scoring of atopic dermatitis (SCORAD) index (r = 0.665, P <.00001), serum lactate dehydrogenese levels (r = 0.696, P= .00001), cosinophil counts (r = 0.381, P = .007), and platelet counts (r = 0.562, P <.0001). Similarly, plasma MDC levels were significantly increased in patients with AD (P <.0001) and showed significant correlations with SCORAD index (r = 0.727, P <.0001), serum lactate dehydrogenese levels (r = 0.861, P <.0001), eosinophil counts (r = 0.505, P = .005), and platelet counts (r = 0.370, P = .01). On treatment, plasma TARC and MDC levels were dramatically decreased in accordance with improved SCORAD scores (P = .0012 and P = .0007, respectively). On the other hand, plasma eotaxin levels did not show any significant increase or correlation with any of the clinical parameters in patients with AD. Conclusion: Platelets from patients with AD contain high levels of TARC. Thus platelets might play an important role in AD pathogenesis by releasing T(H)2-attracting TARC on activation. Furthermore, circulating levels of TARC and MDC, but not those of eotaxin, correlate well with the disease activity of AD.
  • Dual functions of fractalkine/CX3C ligand 1 in trafficking of perforin(+)/granzyme B+ cytotoxic effector lymphocytes that are defined by CX3CR1 expression, M Nishimura, H Umehara, T Nakayama, O Yoneda, K Hieshima, M Kakizaki, N Dohmae, O Yoshie, T Imai, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 168(12), 6173 - 6180, Jun. 2002 , Refereed
    Summary:Fractalkine/CX3C ligand 1 and its receptor CX3CR1 are known to mediate both cell adhesion and cell migration. Here we show that CX3CR1 defines peripheral blood cytotoxic effector lymphocytes commonly armed with intracellular perforin and granzyme B, which include NK cells, gammadelta T cells, and terminally differentiated CD8(+) T cells. In addition, soluble fractalkine preferentially induced migration of cytotoxic effector lymphocytes. Furthermore, interaction of cytotoxic effector lymphocytes with membrane-bound fractalkine promoted subsequent migration to the secondary chemokines, such as macrophage inflammatory protein-1beta/CC ligand 4 or IL-8/CXC ligand 8. Thus, fractalkine expressed on inflamed endothelium may function as a vascular regulator for cytotoxic effector lymphocytes, regardless of their lineage and mode of target cell recognition, through its ability to capture them from blood flow and to promote their emigration in response to other chemokines.
  • Frequent expression of CCR4 in adult T-cell leukemia and human T-cell leukemia virus type 1-transformed T cells, O Yoshie, R Fujisawa, T Nakayama, H Harasawa, H Tago, D Izawa, K Hieshima, Y Tatsumi, K Matsushima, H Hasegawa, A Kanamaru, S Kamihira, Y Yamada, BLOOD, BLOOD, 99(5), 1505 - 1511, Mar. 2002 , Refereed
    Summary:Chemokines and chemokine receptors play important roles In migration and tissue localization of various lymphocyte subsets. Here, we report the highly frequent expression of CCR4 In adult T-cell leukemia (ATL) and human T-cell leukemia virus type 1 (HTLV-1)-immortalized T cells. Flow cytometric analysis revealed that ATL and HTLV-1-immortalized T-cell lines consistently expressed CCR4. Inducible expression of HTLV-1 transcriptional activator tax in a human T-cell line Jurkat did not, however, up-regulate CCR4 mRNA. In vitro Immortalization of peripheral blood T cells led to preferential outgrowth of CD4(+) T cells expressing CCR4. We further demonstrated highly frequent expression of CCR4 in fresh ATL cells by (1) reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of CCR4 expression in peripheral blood mononuclear cells (PBMCs) from patients with ATL and healthy controls; (2) flow cytometric analysis of CCR4-expressing cells in PBMCs from patients with ATL and healthy controls; (3) CCR4 staining of routine blood smears from patients with ATL; and (4) an efficient migration of fresh ATL cells to the CCR4 ligands, TARC/CCL17 and MDC/CCL22, in chemotaxis assays. Furthermore, we detected strong signals for CCR4, TARC, and MDC in ATL skin lesions by RT-PCR. Collectively, most ATL cases have apparently derived from CD4+ T cells expressing CCR4. It is now known that circulating CCR4(+) T cells are mostly polarized to Th2 and also contain essentially all skin-seeking memory T cells. Thus, HTLV-1-infected CCR4+ T cells may have growth advantages by deviating host immune responses to Th2. CCR4 expression may also account for frequent infiltration of ATL into tissues such as skin and lymph nodes. (C) 2002 by The American Society of Hematology.
  • Human B cells immortalized with Epstein-Barr virus upregulate CCR6 and CCR10 and downregulate CXCR4 and CXCR5, T Nakayama, R Fujisawa, D Izawa, K Hieshima, K Takada, O Yoshie, JOURNAL OF VIROLOGY, JOURNAL OF VIROLOGY, 76(6), 3072 - 3077, Mar. 2002 , Refereed
    Summary:Compared to peripheral blood resting B cells, Epstein-Barr virus (EBV)-immortalized B cells consistently express CCR6 and CCR10 at high levels and CXCR4 and CXCR5 at low levels. Accordingly, these cells vigorously responded to the ligands of CCR6 and CCR10 but not to those of CXCR4 and CXCR5. In a human EBV-negative B-cell line, BJAB, stable expression of EBNA2 upregulated CCR6, while stable expression of EBNA2 as well as LMP1 downregulated CXCR4. On the other hand, upregulation of CCR10 or downregulation of CXCR5 was not induced in BJAB by stable expression of EBNA2 or LMP1. Thus, these changes may be due to a plasmablast-like stage of B-cell differentiation fixed by EBV immortalization. EBV-infected B cells in infectious mononucleosis are known to avoid germinal centers and accumulate under the mucosal surfaces. EBV-associated opportunistic lymphomas also tend to occur in extranodal sites. These preferred sites of in vivo localization are consistent with the unique profile of chemokine receptor expression exhibited by EBV-immortalized B cells.
  • Proinflammatory cytokines induce liver and activation-regulated chemokine/macrophage inflammatory protein-3 alpha/CCL20 in mucosal epithelial cells through NK-kappa B, S Fujiie, K Hieshima, D Izawa, T Nakayama, R Fujisawa, H Ohyanagi, O Yoshie, INTERNATIONAL IMMUNOLOGY, INTERNATIONAL IMMUNOLOGY, 13(10), 1255 - 1263, Oct. 2001 , Refereed
    Summary:Liver and activation-regulated chemokine (LARC)/CCL20 is expressed by surface-lining epithelial and epidermal cells, and is likely to link innate and acquired immunity by attracting immature dendritic cells, effector memory T cells and B cells via CCR6. Here we examined the mechanism of LARC expression in epithelial-type cells. Either IL-1 beta or tumor necrosis factor (TNF)-alpha strongly induced LARC mRNA in intestinal cell lines Caco-2 and T84, while both were effective on HEK 293T cells. Induction of LARC was also demonstrated in the intestinal epithelium of BALB/c mice upon treatment with IL-1 alpha or TNF-alpha. Transient transfection assays using murine LARC promoter-reporter constructs identified a region essential for IL-1 beta, or TNF-alpha -induced promoter activation in Caco-2 and 293T cells. Using site-directed mutagenesis, we demonstrated that an NF-kappaB site located between -96 and -87 bp upstream from the transcriptional start site was both necessary and sufficient for IL-1 beta- or TNF-alpha -induced promoter activation in Caco-2 and 293T cells. Electrophoretic mobility shift assays demonstrated that p50/p65 heterodimer and p65 homodimer of NF-kappaB bound to this site in 293T cells upon treatment with IL-1 beta and TNF-alpha, and p50/p65 heterodimer bound to this site in Caco-2 cells upon treatment with IL-1 beta. Co-expression of constitutively active p65 strongly activated the promoter construct carrying the intact NF-kappaB site in 293T and Caco-2 cells. Collectively, LARC expression in intestinal epithelial-type cells is induced by proinflammatory cytokines such as IL-1 and TNF-alpha primarily through activation of NF-kappaB.
  • Human CC chemokine liver-expressed chemokine/CCL16 is a functional ligand for CCR1, CCR2 and CCR5, and constitutively expressed by hepatocytes, H Nomiyama, K Hieshima, T Nakayama, T Sakaguchi, R Fujisawa, S Tanase, H Nishiura, K Matsuno, H Takamori, Y Tabira, T Yamamoto, R Miura, O Yoshie, INTERNATIONAL IMMUNOLOGY, INTERNATIONAL IMMUNOLOGY, 13(8), 1021 - 1029, Aug. 2001 , Refereed
    Summary:Liver-expressed chemokine (LEC)/CCL16 is a human CC chemokine selectively expressed in the liver. Here, we investigated its receptor usage by calcium mobilization and chemotactic assays using mouse L1.2 pre-B cell lines stably expressing a panel of 12 human chemokine receptors. At relatively high concentrations, LEG induced calcium mobilization and chemotaxis via CCR1 and CCR2. LEG also induced calcium mobilization, but marginal chemotaxis via CCR5. Consistently, LEG was found to bind to CCR1, CCR2 and CCR5 with relatively low affinities. The binding of LEG to CCR8 was much less significant. In spite of its binding to CCR5, LEG was unable to inhibit infection of an R5-type HIV-1 to activated human peripheral blood mononuclear cells even at high concentrations. In human liver sections, hepatocytes were strongly stained by anti-LEC antibody. HepG2, a human hepatocarcinoma cell line, was found to constitutively express LEC. LEG was also present in the plasma samples from healthy adult donors at relatively high concentrations (0.3-4 nM). Taken together, LEG is a new low-affinity functional ligand for CCR1, CCR2 and CCR5, and is constitutively expressed by liver parenchymal cells. The presence of LEG in normal plasma at relatively high concentrations may modulate inflammatory responses.
  • Macrophage-derived chemokine (MDC/CCL22) and CCR4 are involved in the formation of T lymphocyte-dendritic cell clusters in human inflamed skin and secondary lymphoid tissue, F Katou, H Ohtani, T Nakayama, K Ono, K Matsushima, A Saaristo, H Nagura, O Yoshie, K Motegi, AMERICAN JOURNAL OF PATHOLOGY, AMERICAN JOURNAL OF PATHOLOGY, 158(4), 1263 - 1270, Apr. 2001 , Refereed
    Summary:Our previous study demonstrated formation of T cell-dendritic cell (DC) clusters in inflamed dermis of intraorally autotransplanted skin flaps. Such T cell-DC clusters are supposed to be important for close interactions between T cells and DCs including the specific antigen presentation. Here we show the involvement of the macrophage-derived chemokine (MDC/CCL22) and its specific receptor CC chemokine receptor 4 (CCR4) in the formation of T cell-DC clusters. Reverse transcriptase-polymerase chain reaction analysis revealed high levels of mRNA expression for MDC and CCR4 in inflamed skin and neck lymph nodes (LNs), but not In normal skin. Immunohistochemically, MDC+ cells and CCR4(+) cells were mainly located within the T cell-DC clusters both in the dermis of inflamed skin and the T cell area of LNs. MDC+ cells were identified to be DCs both in inflamed skin and LNs. The majority of CCR4+ cells were CD4(+) T cells, accounting for approximately one-third of total CD4(+) T cells in the inflamed skin. Our data suggest that the MDC-CCR4 system plays an important role in the formation of T cell-DC clusters both in inflamed skin and LNs.
  • Intervention of thymus and activation-regulated chemokine attenuates the development of allergic airway inflammation and hyperresponsiveness in mice, S Kawasaki, H Takizawa, H Yoneyama, T Nakayama, R Fujisawa, M Izumizaki, T Imai, O Yoshie, Homma, I, K Yamamoto, K Matsushima, JOURNAL OF IMMUNOLOGY, JOURNAL OF IMMUNOLOGY, 166(3), 2055 - 2062, Feb. 2001 , Refereed
    Summary:Thymus- and activation-regulated chemokine (TARC; CCL17) is a lymphocyte-directed CC chemokine that specifically chemoattracts CC chemokine receptor 4-positive (CCR4(+)) Th2 cells. To establish the pathophysiological roles of TARC in vivo, we investigated here whether an mAb against TARC could inhibit the induction of asthmatic reaction in mice elicited by OVA, TARC was constitutively expressed in the lung and was up-regulated in allergic inflammation. The specific Ab against TARC attenuated OVA-induced airway eosinophilia and diminished the degree of airway hyperresponsiveness with a concomitant decrease in Th2 cytokine levels. Our results for the first time indicate that TARC is a pivotal chemokine for the development of Th2-dominated experimental allergen-induced asthma with eosinophilia and AHR. This study also represents the first success in controlling Th2 cytokine production in vivo by targeting a chemokine.
  • Inducible expression of a CC chemokine liver- and activation-regulated chemokine (LARC)/macrophage inflammatory protein (MIP)-3 alpha/CCL20 by epidermal keratinocytes and its role in atopic dermatitis, T Nakayama, R Fujisawa, H Yamada, T Horikawa, H Kawasaki, K Hieshima, D Izawa, S Fujiie, T Tezuka, O Yoshie, INTERNATIONAL IMMUNOLOGY, INTERNATIONAL IMMUNOLOGY, 13(1), 95 - 103, Jan. 2001 , Refereed
    Summary:Liver- and activation-regulated chemokine (LARC)/macrophage inflammatory protein (MIP)3 alpha/ CCL20 is a CC chemokine which is constitutively expressed by follicle-associated epithelial cells in the mucosa, and attracts cells expressing CCR6 such as immature dendritic cells and alpha (4)beta (high)(7) intestine-seeking memory T cells. Here, we examine LARC/CCL20 expression in the skin. LARC/CCL20 mRNA and protein were induced in primary human keratinocytes upon stimulation with proinflammatory cytokines such as IL-1 alpha and tumor necrosis factor (TNF)-alpha, In mice, intradermal injection of IL-l a and TNF-a rapidly induced a local accumulation of transcripts for LARC/CCL20 and its receptor CCR6 with a lag of several hours in the latter. In humans, immunostaining of LARC/CCL20 was weak if any in normal skin tissues but strongly augmented in lesional skin tissues with atopic dermatitis. Furthermore, massive infiltration of cells with markers such as CD1a, CD3 or HLA-DR was present in atopic skin lesions. Many infiltrating cells were also found to be CCR6(+) by a newly generated monoclonal anti-CCR6, However, Langerhans cells residing within the epidermis were hardly stained by anti-CCR6 in normal and atopic skin tissues. Furthermore, plasma levels of LARC/CCL20 were found to be elevated in patients with atopic dermatitis. Collectively, our results suggest that epidermal keratinocytes produce LARC/CCL20 upon stimulation with proinflammatory cytokines such as IL-1 alpha and TNF-alpha, and attract CCR6-expressing immature dendritic cells and memory/effector T cells into the dermis of inflamed skin such as atopic dermatitis. LARC/CCL20 may not, however, play a major role in homeostatic migration of Langerhans cells into the skin.
  • Molecular cloning of a novel CC chemokine, interleukin-11 receptor alpha-locus chemokine (ILC), which is located on chromosome 9p13 and a potential homologue of a CC chemokine encoded by molluscum contagiosum virus, Ishikawa-Mochizuki, I, M Kitaura, M Baba, T Nakayama, D Izawa, T Imai, H Yamada, K Hieshima, R Suzuki, H Nomiyama, O Yoshie, FEBS LETTERS, FEBS LETTERS, 460(3), 544 - 548, Nov. 1999 , Refereed
    Summary:Molluscum contagiosum virus (MCV) encodes a CC chemokine MC148R which is likely to have been acquired from the host. By a homology search employing MC148R as a probe, we hale identified a novel CC chemokine whose gene exists next to the IL-11 receptor alpha (IL-11R alpha) gene in both humans and mice. Thus, this chemokine maps to chromosome 9p13 in humans where IL-11R alpha has been assigned, We term this novel chemokine IL-11R alpha-locus chemokine (ILC), ILC has the highest homology to MC148R among the known human CC chemokines. Furthermore, ILC is strongly and selectively expressed in the skin where infection of MCV also takes place, Thus, ILC is likely to be the original chemokine of MC148R. (C) 1999 Federation of European Biochemical Societies.
  • Molecular cloning of mXCR1, the murine SCM-1/lymphotactin receptor, T Yoshida, D Izawa, T Nakayama, K Nakahara, M Kakizaki, T Imai, R Suzuki, M Miyasaka, O Yoshie, FEBS LETTERS, FEBS LETTERS, 458(1), 37 - 40, Sep. 1999 , Refereed
    Summary:Single C motif-1 (SCM-1)/lymphotactin is a C-type member of the chemokine superfamily, Previously, we identified its specific receptor XCR1. Here we isolated the murine homologue of XCR1 (mXCR1). To demonstrate its biological activity, we produced recombinant mouse SCM-1 by the baculovirus expression system, B300-19 murine pre-B cells expressing mXCR1 responded to mSCM-1 in chemotactic and calcium-mobilization assays. mXCR1 mRNA was weakly expressed in spleen and lung of normal C57BL/6 mice. In spleen, CD8(+) cells and NK1.1(+) cells were found to express mXCR1. Identification of mXCR1 will now allow us to study the role of this unique cytokine system in the mouse models of inflammation and immunity, (C) 1999 Federation of European Biochemical Societies.
  • Ca2+-dependent interaction of N-copine, a member of the two C2 domain protein family, with OS-9, the product of a gene frequently amplified in osteosarcoma, T Nakayama, T Yaoi, G Kuwajima, O Yoshie, T Sakata, FEBS LETTERS, FEBS LETTERS, 453(1-2), 77 - 80, Jun. 1999 , Refereed
    Summary:N-copine is a novel two C2 domain protein that shows Ca2+-dependent phospholipid binding and membrane association. By using yeast two-hybrid assays, we identified OS-9 as a protein capable of interacting with N-copine, We further revealed that the second C2 domain of N-copine bound with the carboxy-terminal region of OS-9, Their interaction in vivo nas also confirmed by co-immunoprecipitation from 293E cells co-expressing transfected N-copine and OS-9, In vitro binding assays showed that this interaction was Ca2+-dependent. By Northern blot analysis, N-copine and OS-9 were co-expressed in the same regions of human brain. These results reveal that OS-9 is a potential target of N-copine. (C) 1999 Federation of European Biochemical Societies.
  • Localization and subcellular distribution of N-copine in mouse brain, T Nakayama, T Yaoi, G Kuwajima, JOURNAL OF NEUROCHEMISTRY, JOURNAL OF NEUROCHEMISTRY, 72(1), 373 - 379, Jan. 1999 , Refereed
    Summary:N-Copine is a novel protein with two C2 domains. Its expression is brain specific and up-regulated by neuronal activity such as kainate stimulation and tetanus stimulation evoking hippocampal CA1 long-term potentiation. We examined the localization and subcellular distribution of N-copine in mouse brain. In situ hybridization analysis showed that N-copine mRNA was expressed exclusively in neurons of the hippocampus and in the main and accessory olfactory bulb, where various forms of synaptic plasticity and memory formation are known to occur. In immunohistochemical analyses, N-copine was detected mainly in the cell bodies and dendrites in the neurons, whereas presynaptic proteins such as synaptotagmin I and rab3A were detected in the regions where axons pass through. In fractionation experiments of brain homogenate, N-copine was associated with the membrane fraction in the presence of Ca2+ but not in its absence. As a GST-fusion protein with the second C2 domain of N-copine showed Ca2+-dependent binding to phosphatidylserine, this domain was considered to be responsible for the Ca2+-dependent association of N-copine with the membrane. Thus, N-copine may have a role as a Ca2+ sensor in postsynaptic events, in contrast to the known roles of "double C2 domain-containing proteins," including synaptotagmin I, in presynaptic events.
  • N-copine: a novel two C2-domain-containing protein with neuronal activity-regulated expression, T Nakayama, T Yaoi, M Yasui, G Kuwajima, FEBS LETTERS, FEBS LETTERS, 428(1-2), 80 - 84, May 1998 , Refereed
    Summary:Neuronal activity is often associated with changes in gene expression. By a two-dimensional cDNA-display system, restriction landmark cDNA scanning, we identified a novel gene whose expression in the hippocampus was up-regulated by kainate stimulation. The mRNA expression was detected only in brain and up-regulated by the stimulation making CA3-CA1 long-term potentiation. The encoded protein contains two copies of C2-domain, known as the Ca2+-binding domain of PKC-gamma and shows 49% identity with human copine I. We designated this protein N-copine (neuronal-copine), N-copine may have a role in synaptic plasticity. (C) 1998 Federation of European Biochemical Societies.

Conference Activities & Talks

  • Induction of FosB by Tax induces CCL22 in HTLV-1-infected T cells, leading to HTLV-1 transmission to CCR4+ T cells.,   2012 10
  • Fra-2-SOX4 oncogenic cascade plays a critical role in cell growth of CCR4+ mature T-cell malignancies, JSICR-MMCB2011,   2011 05 , JSICR-MMCB2011
  • Common oncogenic role of Fra-2 in mature T-cell malignancies expressing CCR4., Gordon Reserch Conferences,   2010 06 , Gordon Reserch Conferences
  • Identification of Fra-2-SOX4 oncogenic cascade in CCR4+ mature T-cell malignancies, 14th International Congress of Immunology,   2010 , 14th International Congress of Immunology
  • Common oncogenic role of FRA-2 in mature T-cell lymphoma expressing CCR4., International Association of Inflammation Societie T-cell Lymphoma Forum,   2010 01 , International Association of Inflammation Societie T-cell Lymphoma Forum
  • Eotaxin-3/CCL26 interacts with CX3CR1 and induces killer lymphocyte migration.,   2009 12
  • CXCR7 induced by Tax promotes proliferation of HTLV-1 infected T cells, The 9th World Congress on Inflammation,   2009 07 , The 9th World Congress on Inflammation
  • The role of Fra-2 and c-Myb oncogenic cascade in CD4+ mature T-cell lymphomas,   2009
  • Fra-2 and c-Myb oncogenic cascade is involved in cell growth in CD4+ mature T-cell lymphomas,   2009
  • Functional analysis of the Th2-related transcription factor c-Maf in HTLV-1 infection,   2008 10
  • Aberrantly expressed Fra-2 promotes CCR4 expression and cell proliferation in adult T-cell leukemia., International Investigative Dermatology,   2008 05 , International Investigative Dermatology
  • Induction of CCR10 expression by 1,25-Dihydroxyvitamin D3 in Terminally Differentiating Human B Cells, Keystone Symposia 2008,   2008 01 , Keystone Symposia 2008
  • Role of Fra-2/JunD heterodimer in CCR4 expression and cell proliferation in adult T-cell leulemia,   2007 11
  • Induction of CCR10 expression by 1,25-Dihydroxyvitamin D3 in Terminally Differentiating Human B Cells,   2007 11
  • HTLV-1 Tax-induced CCL22 promotes primary infection of HTLV-1 to peripheral blood CCR4+CD4+ T cells,   2007 10
    Summary:We examined the effect of Tax on expression of CCL22, a ligand of CCR4. Most Tax-expressing HTLV-1+ T cell lines produced a large amount of CCL22 in their culture supernatants. Transient expression of Tax also induced CCL22. Knockdown of Tax gene by siRNA resulted in a reduction of CCL22 mRNA. These results indicate CCL22 is one of the target genes of Tax. Culture supernatants from the HTLV-1+ T cell lines selectively attracted primary CCR4+CD4+ T cells in chemotaxis assays. The migration was blocked by pretreatment of the supernatants with anti-CCL22 Ab or pretreatment of PBMCs with a CCR4-specific antagonist. Further, HTLV-1+ T cell lines showed significant CCR4-dependent cell-to-cell interactions with sorted primary CCR4+CD4+ T cells, and anti-CCL22 Ab inhibited HTLV-1 primary infection. These results indicate that HTLV-1-infected T cells produce CCL22 upon expression of Tax and selectively recruit CCR4+CD4+ T cells, resulting in preferential transmission of HTLV-1 to these cells.
  • Heterodimer formation of Fra-2 and JunD promotes CCR4 expression and cell proliferation in adult T-cell leukemia,   2007 10
  • Potential role of CCR9 in gastrointestinal involvement of ATL cells., IMMUNOLOGY 2007, The American Association of Immunologists,   2007 05 , IMMUNOLOGY 2007, The American Association of Immunologists
  • Involvement of fra-2 and JunD in CCR4 expression and cell proliferation in adult T cell leukemia, 13th International Conference on Human Retrovirology HTLV and Related Viruses,   2007 05 , 13th International Conference on Human Retrovirology HTLV and Related Viruses
  • HTLV-1 Tax-induced CCL22 promotes cell-cell interactions between HTLV-1-infected cells and peripheral blood CCR4+CD4+ T cells, The 13th International Conference on Human Retrovirology; HTLV and Related Viruses,   2007 05 , The 13th International Conference on Human Retrovirology; HTLV and Related Viruses
    Summary:Most Tax+ HTLV-1+ T cell lines produced large amounts of CCL22 in their culture supernatants. Transient Tax expression also expressed CCL22. siRNA-induced knockdown of Tax resulted in a significant reduction of CCL22 mRNA in Tax+ cells. These results indicated CCL22 is one of the target genes of Tax. By using PBMCs from healthy donors in chemotaxis assays, it was demonstrated that culture supernatants of Tax+ cells strongly mobilized CCR4+CD4+ T cells. The migration was blocked by pretreatment of the supernatants with anti-CCL22 neutralizing antibody or pretreatment of the PBMCs with a CCR4-specific inhibitor. Further, Tax+ cells showed significant CCR4-dependent cell-cell interactions with the sorted CCR4+CD4+ T cells. These results imply that HTLV-1+ T cells, which were vertically transmitted from the breast milk of a carrier mother, produced CCL22 upon expression of Tax and preferentially recruited CCR4+CD4+ T cells in the child.
  • CCL27 transgenic mice showed enhanced contact hypersensitivity reaction to repeated challenges with fluorescein isothiocyanate., Society for Investigative Dermatology Annual Meeting,   2007 05 , Society for Investigative Dermatology Annual Meeting
  • Induction of chemokine in human corneal epithelial cells and corneal keratocytes,   2004 11
  • roles of chemokine in herpetic keratitis,   2004 07
  • Corneal epithelial cells and stromal keratocytes efficiently produce CC chemokine-ligand(CCL20) and attract inflammatory cells in mouse herpetic stromal keratitis,   2004 04
  • MEC/CCL28 Has Dual Functions in Mucosal Immunity as a Chemoattractant with Broad-Spectrum Antimicrobial Activity,   2003 06
  • CCL28 has a dual role in mucosal immunity as chemokine with a broad-spectrum antimicrobial activity., Keystone Symposia 2003,   2003 01 , Keystone Symposia 2003
  • Anti-tumor effect by a CC chemokine, CCL27, introduces into tumor cells through a recombinant adenovirus vector,   2003

Misc

  • Chemokine receptors and cell migration, 松尾一彦, 中山隆志, 日本血栓止血学会誌, 30, 4, 610‐618(J‐STAGE),   2019 , 10.2491/jjsth.30.610, https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201902244635586992
  • Search for chemokine receptor CCR3 selective antagonist as anti-allergic principles from mace, the arils of Myristica fragrans, Morikawa Toshio, Matsuo Kazuhiko, Hachiman Ikuko, Ninomiya Kiyofumi, Muraoka Osamu, Nakayama Takashi, Symposium on the Chemistry of Natural Products, symposium papers, 56, 0,   2018 07 19 , http://ci.nii.ac.jp/naid/130007399508
    Summary:<p>Chemokines play pivotal roles in health and disease by controlling migration and tissue localization of specific types of cells expressing their cognate receptors. They are grouped into four subfamilies (CXC, CC, CX3C, and XC) by the structural motif of the N-terminal conserved cysteine residues. Among the CC chemokine receptors, CCR3 is selectively expressed on eosinophils, basophils, and some Th2 cells, and plays an essential role in the pathogenesis of allergic diseases by modifying the kinetics of these cells. In the course of our characterization studies on anti-allergic principles from natural resources, we found that a methanol extract from mace, the arils of Myristica fragrans, showed inhibitory effect on eotaxin/CCL11-induced cell migration in L1.2 cells expressing CCR3 at a concentration of 100 mg/mL. Among the isolates from the extract, three new neolignans 3 (EC<sub>50</sub> = 1.62 mM), 4 (1.47 mM), and 5 (1.35 mM) were identified as the constituents responsible for the activity, and these activities were equivalent to that of SB328437 (EC<sub>50</sub> = 0.78 mM), a CCR3 selective antagonist. These results indicated that the neolignan constituents (3–5) are potential candeidates for the new anti-allergic agents.</p>
  • メース(Myristica fragrans,仮種皮)のケモカイン受容体CCR3アンタゴニスト様作用を指標とした抗アレルギー作用成分の探索, 森川敏生, 森川敏生, 八幡郁子, 松尾一彦, 西田枝里子, 二宮清文, 二宮清文, 義江修, 村岡修, 村岡修, 中山隆志, 食品薬学シンポジウム講演要旨集, 7th, 134‐136,   2017 10 10 , http://jglobal.jst.go.jp/public/201702219970879037
  • がんゲノム医療の実装と遺伝性疾患の責任遺伝子探索・創薬から予防までのトータルケアー, 西尾 和人, 田村 和朗, 西郷 和真, 杉浦 麗子, 中山 隆志, 奥野 清隆, 菰池 佳史, 万代 昌紀, 竹村 司, 伊藤 彰彦, 大磯 直毅, 浮田 真沙世, 坂井 和子, 日本遺伝カウンセリング学会誌, 38, 2, 35, 36,   2017 05
  • CCR4阻害剤はTregの筋肉組織への遊走を阻害することでワクチン効果を向上させる, 東山慎太郎, 松尾一彦, 松永奈緒子, 山田祐毅, 西脇敬二, 義江修, 中山隆志, 日本薬学会年会要旨集(CD-ROM), 137th, ROMBUNNO.26PB‐am071S,   2017 , http://jglobal.jst.go.jp/public/201702227068290497
  • ケモカイン受容体CCR4の欠損はメラノーマ担癌モデルマウスの病態を増悪させる, 高橋周平, 松尾一彦, 小山篤, 西脇敬二, 義江修, 中山隆志, 日本薬学会年会要旨集(CD-ROM), 137th, ROMBUNNO.26PB‐am068S,   2017 , http://jglobal.jst.go.jp/public/201702242388466738
  • CCR4欠損マウスおよびCCR6欠損マウスを用いたイミキモド誘発性乾癬の解析, 伊藤茉奈, 松尾一彦, 長沼孝典, 西脇敬二, 義江修, 中山隆志, 日本薬学会年会要旨集(CD-ROM), 137th, ROMBUNNO.26PB‐am070S,   2017 , http://jglobal.jst.go.jp/public/201702254581423306
  • ケモカイン受容体CCR4の欠損はアトピー様皮膚炎の病態を改善させる, 木村勇太, 松尾一彦, 小森悠平, 畑中翔太, 西脇敬二, 義江修, 中山隆志, 日本薬学会年会要旨集(CD-ROM), 137th, ROMBUNNO.26PB‐am069S,   2017 , http://jglobal.jst.go.jp/public/201702260154362718
  • CS分析(Customer Satisfaction analysis)による薬剤師のためのフィジカルアセスメント講習会の評価と改善, 大鳥 徹, 井上 知美, 細見 光一, 中川 博之, 高島 敬子, 近藤 尚美, 高田 亜美, 伊藤 栄次, 中山 隆志, 和田 哲幸, 石渡 俊二, 前川 智弘, 船上 仁範, 中村 真也, 窪田 愛恵, 平出 敦, 松山 賢治, 西田 升三, 社会薬学, 35, 2, 94, 101,   2016 12 , 10.14925/jjsp.35.2_94
    Summary:フィジカルアセスメント講習会に参加した143名を対象にアンケートを実施した。CS分析(Customer Satisfaction analysis)とテキストマイニングにより検討した。男性が39%、女性が61%と女性が多かった。勤務先は保険調剤薬局が38%、病院が62%となっており、病院勤務者が多かった。改善度は、数値が大きいほど改善の優先順位は高くなるが、改善度の値が大きい順に五つ項目を上げると、「心電図の読み方」「呼吸音の聴診」「SBAR」「時間は適当でしたか」「心音の聴診」であった。CSグラフにおいて、優先的に改善する必要がある項目がプロットされる「要改善項目」(第4象限)として抽出されてきたのは、「呼吸音の聴診」、「SBAR」であった。「心音・腸音の聴診」、「心電図の読み方」、「血圧測定」は評価が低く、総合評価への影響度も低い項目である「改善検討項目」として第3象限にプロットされた。「講義内容」、「症例に興味が持てたか」、「フィジカルアセスメントモデルの機能」、「時間は適当でしたか」は評点が高く、総合評価への影響度も高い項目である「重要維持項目」として第1象限にプロットされた。
  • ケモカイン受容体を標的とした天然由来シーズの探索研究(2):ロウバイカ(Chimonanthus praecox花蕾部)のCCR6およびCCR7アンタゴニスト活性成分, 森川敏生, 松尾一彦, 奥川修平, 奥川修平, 北川仁一朗, 北川仁一朗, 二宮清文, 中西勇介, 村岡修, 村岡修, 中山隆志, 中山隆志, 日本生薬学会年会講演要旨集, 63rd, 146,   2016 08 25 , http://jglobal.jst.go.jp/public/201602251743247114
  • 和漢薬ライブラリーを利用したケモカイン受容体CCR3及びCCR4のアンタゴニスト成分の探索, 北田 卓也, 松尾 一彦, 小泉 桂一, 義江 修, 中山 隆志, 日本薬学会年会要旨集, 135年会, 3, 167, 167,   2015 03
  • メースの機能性成分(4);ケモカイン受容体CCR3選択的アゴニスト作用成分の探索, 森川敏生, 八幡郁子, 松尾一彦, 二宮清文, 村岡修, 中山隆志, 日本生薬学会年会講演要旨集, 61st, 118,   2014 08 27 , http://jglobal.jst.go.jp/public/201402294364708616
  • Search for chemokine receptor CCR3 selective antagonist as anti-allergic principles from mace, the arils of Myristica fragrans, Morikawa Toshio, Matsuo Kazuhiko, Hachiman Ikuko, Ninomiya Kiyofumi, Muraoka Osamu, Nakayama Takashi, Symposium on the Chemistry of Natural Products, symposium papers, 56, 0,   2014 , http://ci.nii.ac.jp/naid/130007399508
    Summary:<p>Chemokines play pivotal roles in health and disease by controlling migration and tissue localization of specific types of cells expressing their cognate receptors. They are grouped into four subfamilies (CXC, CC, CX3C, and XC) by the structural motif of the N-terminal conserved cysteine residues. Among the CC chemokine receptors, CCR3 is selectively expressed on eosinophils, basophils, and some Th2 cells, and plays an essential role in the pathogenesis of allergic diseases by modifying the kinetics of these cells. In the course of our characterization studies on anti-allergic principles from natural resources, we found that a methanol extract from mace, the arils of Myristica fragrans, showed inhibitory effect on eotaxin/CCL11-induced cell migration in L1.2 cells expressing CCR3 at a concentration of 100 mg/mL. Among the isolates from the extract, three new neolignans 3 (EC<sub>50</sub> = 1.62 mM), 4 (1.47 mM), and 5 (1.35 mM) were identified as the constituents responsible for the activity, and these activities were equivalent to that of SB328437 (EC<sub>50</sub> = 0.78 mM), a CCR3 selective antagonist. These results indicated that the neolignan constituents (3–5) are potential candeidates for the new anti-allergic agents.</p>
  • 和漢薬の科学基盤 共同研究による先駆的統合的解明 標準和漢薬ライブラリーを用いたケモカイン受容体CCR3およびCCR4アンタゴニストの探索 アレルギー疾患を標的として, 中山 隆志, 田中 宏幸, 松尾 一彦, 小泉 桂一, 義江 修, 日本薬学会年会要旨集, 133年会, 1, 146, 146,   2013 03
  • 形質細胞におけるCCR10発現の活性型ビタミンD3による誘導, 白川愛子, 長久保大輔, 稗島州雄, 中山隆志, 金哲, 義江修, 日本栄養・食糧学会大会講演要旨集, 65th, 105,   2011 04 25 , http://jglobal.jst.go.jp/public/201102204242615128
  • CCR4陽性成熟型T細胞リンパ脚におけるFra‐2‐c‐Myb発癌遺伝子カスケードの役割, 樋口智紀, 中山隆志, 長久保大輔, 重田暁子, 白川愛子, 稗島州雄, 義江修, 日本分子生物学会年会講演要旨集, 32nd, Vol.1, 232,   2009 , http://jglobal.jst.go.jp/public/201002264746169833
  • 大腸癌におけるCXCL16の発現と予後との関連, 北條 荘三, 小泉 桂一, 有田 貴久, 篠原 看奈, 南 貴之, 橋本 伊佐也, 中山 隆志, 櫻井 宏明, 義江 修, 塚田 一博, 済木 育夫, 日本癌学会総会記事, 65回, 337, 338,   2006 09
  • Chemokine receptor CXCR4阻害による胃癌腹膜播種の抑制, 安本 和生, 小泉 桂一, 済木 育夫, 川島 篤弘, 中山 隆志, 義江 修, 高橋 豊, 日本癌学会総会記事, 65回, 455, 455,   2006 09
  • Senile/Age‐Related EBV+B‐Cell Lymphoproliferative Disorder(SLPD)におけるケモカイン発現と腫瘍局所浸潤T細胞の解析, JIN Zhe, 中山隆志, 白川愛子, 長久保大輔, 稗島州雄, 中村栄男, 義江修, 日本癌学会学術総会記事, 65th, 125,   2006 08 28 , http://jglobal.jst.go.jp/public/200902272358399675
  • ドーパミンレセプターD3を介したナイーブCD8陽性T細胞の遊走とリンパ節ホーミング, 渡辺賀子, 中山隆志, 長久保大輔, 稗島州雄, 義江修, 日本免疫学会総会・学術集会記録, 35, 166,   2005 11 15 , http://jglobal.jst.go.jp/public/200902219193714815
  • Senile EBV+B‐Cell Lymphoproliferative Disorder(SLPD)におけるケモカイン発現と病態形成, JIN Zhe, 中山隆志, 長久保大輔, 稗島州雄, 義江修, 日本免疫学会総会・学術集会記録, 35, 163,   2005 11 15 , http://jglobal.jst.go.jp/public/200902228046747294
  • 成人T細胞白血病細胞の腸管浸潤におけるケモカイン受容体CCR9の役割, 長久保大輔, JIN Zhe, 中山隆志, 稗島州雄, 義江修, 日本免疫学会総会・学術集会記録, 35, 163,   2005 11 15 , http://jglobal.jst.go.jp/public/200902295385752247
  • CCL21により誘導されるマウス肺癌細胞の転移関連因子の検索およびその受容体CCR7の発現機構の解析, 有田 貴久, 小泉 桂一, 中山 隆志, 稗島 州雄, 義江 修, 櫻井 宏明, 済木 育夫, 日本癌学会総会記事, 64回, 99, 99,   2005 09
  • 破骨細胞分化誘導に伴うCCL22の産生とヒト小細胞肺癌株SBC-5細胞の骨転移機構の解明, 小泉 桂一, 小林 光夫, 中村 エリアネ静, 斉藤 百合花, 中山 隆志, 櫻井 宏明, 亀田 陽一, 義江 修, 済木 育夫, 日本癌学会総会記事, 64回, 168, 169,   2005 09
  • ヒト前立腺癌細胞株におけるDHT-AR誘導によるケモカイン受容体の発現亢進と細胞機能の変化について, 明石 拓也, 有田 貴久, 小泉 桂一, 永川 修, 斎藤 百合花, 篠原 看奈, 南 貴之, 中山 隆志, 義江 修, 櫻井 宏明, 済木 育夫, 布施 秀樹, 日本癌学会総会記事, 64回, 294, 294,   2005 09
  • Chemokine Receptor CXCR4発現胃癌が腹膜播種形成に関与する, 安本 和生, 小泉 桂一, 櫻井 宏明, 済木 育夫, 中山 隆志, 稗島 州雄, 義江 修, 川島 篤弘, 高橋 豊, 磨伊 正義, 日本癌学会総会記事, 64回, 387, 387,   2005 09
  • ヘルペス性眼疾患―DNAから個体まで―, 下村嘉一, 松本長太, 福田昌彦, 丸山耕一, 日比野剛, 桧垣史郎, 宇野直樹, 菅原大輔, 妙中直子, 出合達則, 白根授美, 有村英子, 渡辺敬三, 義江修, 中山隆志, 長久保大輔, ひえ島州雄, 林こう三郎, 臨床眼科, 59, 6, 816, 825,   2005 06 15 , 10.11477/mf.1410100272, http://jglobal.jst.go.jp/public/200902230773170107
  • IL-12とCCL27の併用による抗腫瘍効果増強機構の解明, 杉田 敏樹, 高 建青, 金川 尚子, 飯田 恵介, 本村 吉章, 畑中 豊, 谷 一, 中山 隆志, 義江 修, 水口 裕之, 早川 尭夫, 岡田 直貴, 堤 康央, 弓 忠範, 中川 晋作, 日本薬学会年会要旨集, 125年会, 2, 154, 154,   2005 03
  • EBV感染B細胞におけるケモカイン発現の網羅的解析, 中山隆志, 長久保大輔, 稗島州雄, 義江修, 日本分子生物学会年会プログラム・講演要旨集, 27th, 500,   2004 11 25 , http://jglobal.jst.go.jp/public/200902285263298876
  • ドーパミンレセプターD3を介したCD8陽性T細胞の遊走, 渡辺賀子, 中山隆志, 長久保大輔, ひ島州雄, 義江修, 日本免疫学会総会・学術集会記録, 34, 44,   2004 11 05 , http://jglobal.jst.go.jp/public/200902214478804964
  • IgA抗体産生細胞の腸管粘膜固有層へのホーミングを誘導するケモカインの解析, 稗島州雄, 中山隆志, 長久保大輔, 義江修, 日本免疫学会総会・学術集会記録, 34, 37,   2004 11 05 , http://jglobal.jst.go.jp/public/200902215834091516
  • Liver‐Expressed Chemokine/CCL16のヒスタミンH4レセプターに対する作用と好酸球遊走, 中山隆志, 加藤佳子, 稗島州雄, 長久保大輔, 藤沢隆夫, 義江修, 日本免疫学会総会・学術集会記録, 34, 38,   2004 11 05 , http://jglobal.jst.go.jp/public/200902290825086552
  • ケモカイン発現ベクターの腫瘍内投与による抗腫瘍効果と免疫細胞浸潤, 岡田 直貴, 中川 晋作, 畑中 豊, 谷 洋一, 中山 隆志, 義江 修, 水口 裕之, 早川 高夫, 藤田 卓也, 山本 昌, 日本免疫学会総会・学術集会記録, 34, 215, 215,   2004 11
  • IL-12及びCCL27発現アデノウイルスベクターの併用投与による抗腫瘍効果と免疫系細胞の浸潤, 杉田 敏樹, 畑中 豊, 谷 洋一, 中山 隆志, 義江 修, 水口 裕之, 早川 高夫, 岡田 直貴, 堤 康央, 真弓 忠範, 中川 晋作, 日本免疫学会総会・学術集会記録, 34, 215, 215,   2004 11
  • マウス肺癌のリンパ節転移に対するCCR7の発現とCCL21/Secondary Lymphoid-tissue Chemokine(SLC)の影響, 有田 貴久, 小林 光夫, 小澤 陽子, 大橋 養賢, 中村 エリアネ静, 青塚 保志, 齊藤 百合花, 櫻井 宏明, 中山 隆志, 稗島 州雄, 義江 修, 小泉 桂一, 済木 育夫, 日本癌学会総会記事, 63回, 263, 263,   2004 09
  • リンパ組織移行性を増強した樹状細胞ワクチンの創製, 森直樹, 岡田直貴, 是友良介, 岡田裕香, 中山隆志, 義江修, 水口裕之, 早川尭夫, 中川晋作, 日本薬学会年会要旨集, 124th, 4, 112, 112,   2004 03 05 , http://jglobal.jst.go.jp/public/200902222945713539
  • ケモカイン発現ベクターの腫よう内投与を併用した樹状細胞ワクチンの抗腫よう効果, 佐々木明徳, 岡田直貴, 丹羽正和, 岡田裕香, 中山隆志, 義江修, 畑中豊, 水口裕之, 中川晋作, 日本薬学会年会要旨集, 124th, 4, 112,   2004 03 05 , http://jglobal.jst.go.jp/public/200902249210413685
  • Cytokine, chemokine, and their receptors in allergic inflammation of the skin and its treatment. Control of skin-specific chemokine production., 義江修, 中山隆志, 稗島州雄, 長久保大輔, 免疫アレルギー疾患予防・治療研究事業研究報告書 平成15年度 第2分冊, 66, 70,   2004 , http://jglobal.jst.go.jp/public/200902224553872275
  • ケモカインCCL28/MECの唾液腺における役割, 稗島州雄, 大谷明夫, 伊沢大, 中山隆志, 川崎ゆり, 斎藤卓也, 義江修, 日本唾液腺学会抄録集, 48th, 28, 29,   2003 12 13 , http://jglobal.jst.go.jp/public/200902275816259763
  • 腸管免疫におけるケモカインCCL28/CCR10系の役割, 稗島州雄, 中山隆志, 長久保大輔, 義江修, 川崎ゆり, 花本仁, 日本免疫学会総会・学術集会記録, 33, 228,   2003 11 05 , http://jglobal.jst.go.jp/public/200902263629276397
  • EBV不死化B細胞におけるケモカイン発現とその制御機構の解析, 中山隆志, 長久保大輔, 稗島州雄, 義江修, 日本免疫学会総会・学術集会記録, 33, 231,   2003 11 05 , http://jglobal.jst.go.jp/public/200902275628918784
  • ATLLにおけるケモカインレセプター発現の網羅的解析, 義江修, 中山隆志, 長久保大輔, 稗島州雄, 原沢仁美, 山田恭あき, 日本免疫学会総会・学術集会記録, 33, 232,   2003 11 05 , http://jglobal.jst.go.jp/public/200902292728203628
  • ケモカイン遺伝子導入を併用したDC癌免疫療法プロトコールの有効性評価, 佐々木明徳, 岡田直貴, 中山隆志, 義江修, 畑中豊, 谷洋一, 中川晋作, 早川高夫, 水口裕之, 日本免疫学会総会・学術集会記録, 33, 327, 327,   2003 11 05 , http://jglobal.jst.go.jp/public/200902223996572987
  • CCケモカインレセプター7(CCR7)遺伝子を導入した樹状細胞のin vitroおよびin vivo遊走能, 森直樹, 岡田直貴, 井上恵美子, 中山隆志, 義江修, 中川晋作, 真弓忠範, 早川高夫, 水口裕之, 日本免疫学会総会・学術集会記録, 33, 327,   2003 11 05 , http://jglobal.jst.go.jp/public/200902283158505561
  • 樹状細胞免疫療法の有効性改善を目指したケモカイン発現ベクターの併用, 佐々木明徳, 岡田裕香, 中山隆志, 義江修, 水口裕之, 早川尭夫, 中川晋作, 真弓忠範, 山本昌, Drug Deliv Syst, 18, 3, 248, 248,   2003 05 10 , http://jglobal.jst.go.jp/public/200902256841575772
  • ケモカイン発現RGDファイバーミュータントアデノウイルスベクターの樹状細胞免疫療法への応用に関する基礎的検討, 佐々木明徳, 岡田直貴, 大久保米起, 岡田裕香, 中山隆志, 水口裕之, 中川晋作, 藤田卓也, 山本昌, 日本薬学会年会要旨集, 123rd, 4, 81, 81,   2003 03 05 , http://jglobal.jst.go.jp/public/200902236884993313
  • ヒト非小細胞肺癌(NSCLC)に対するSecondary Lymphoid-tissue Chemokine(SLC)のリンパ節転移亢進因子としての可能性, 小泉 桂一, 小澤 陽子, 大橋 養賢, 中村 エリアネ静, 中山 隆志, 櫻井 宏明, 義江 修, 済木 育夫, 日本癌学会総会記事, 61回, 110, 110,   2002 10
  • 大腸癌先進部におけるケモカイン及びケモカインレセプターの発現, 武者 宏昭, 溝井 賢幸, 中山 隆志, 大谷 明夫, 椎葉 健一, 義江 修, 佐々木 巖, 日本癌学会総会記事, 61回, 360, 361,   2002 10
  • 大腸癌先進部におけるケモカイン及びケモカインレセプターの発現, 武者 宏昭, 椎葉 健一, 溝井 賢幸, 中山 隆志, 大谷 明夫, 義江 修, 佐々木 巖, 日本免疫学会総会・学術集会記録, 32, 162, 162,   2002 10
  • ケモカインを用いた癌免疫療法の基礎的検討, 津田育宏, 中川晋作, 塚田有希子, 水口裕之, 早川尭夫, 中山隆志, 義江修, 真弓忠範, 薬剤学, 61, Supplement, 74, 74,   2001 03 05 , http://jglobal.jst.go.jp/public/200902102259345587
  • Chemokine Receptor Antagonists Ameliorate Experimental Autoimmune Encephalomyelitis., Katsuichi Miyamoto, Kota Moriguchi, Rino Ueno, Takashi Nakayama, Susumu Kusunoki, MULTIPLE SCLEROSIS JOURNAL, 22, 3, 433, 433,   2016 03
  • Tax-inducible production of CC chemokine ligand 22 by human T cell leukemia virus type 1 (HTLV-1)-infected T cells promotes preferential transmission of HTLV-1 to CDR4-expressing CD4(+) T cells (vol 180, pg 931, 2008), K. Hieshima, D. Nagakubo, T. Nakayama, A. -K. Shirakawa, Z. Jin, O. Yoshie, JOURNAL OF IMMUNOLOGY, 180, 12, 8470, 8470,   2008 06
  • HTLV-1 tax-induced CCL22 promotes cell-cell interactions between HTLV-1-infected cells and peripheral blood CCR4(+) CD4(+) T cells, Kunio Hieshima, Daisuke Nagakubo, Takashi Nakayama, Aiko-Konno Shirakawa, Osamu Yoshie, AIDS RESEARCH AND HUMAN RETROVIRUSES, 23, 4, 646, 646,   2007 04
  • Involvement of fra-2 and JunD in CCR4 expression and cell proliferation in adult T cell leukemia, Takashi Nakayama, Kunio Hieshima, Zhe Jin, Daisuke Nagakubo, Aiko-Konno Shirakawa, Yasuaki Yamada, Masahiro Fujii, Osamu Yoshie, AIDS RESEARCH AND HUMAN RETROVIRUSES, 23, 4, 642, 643,   2007 04
  • CCL27 transgenic mice showed enhanced contact hypersensitivity reaction to repeated challenges with fluorescein isothiocyanate, S. Kagami, H. Saeki, Y. Tsunemi, K. Nakamura, T. Nakayama, O. Yoshie, M. Komine, K. Tamaki, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 127, S113, S113,   2007 04
  • Potential role of CCR9 in gastrointestinal involvement of ATL cells, Daisuke Nagakubo, Zhe Jin, Kunio Hieshima, Takashi Nakayama, Aiko-Konno Shirakawa, Osamu Yoshie, JOURNAL OF IMMUNOLOGY, 178,   2007 04
  • CXCR4 in peritoneal carcinomatosis of gastric carcinoma (vol 66, pg 2181, 2006), K Yasumoto, K Koizumi, A Kawashimu, Y Saitoh, Y Arita, K Shinohara, T Minami, T Nakayama, H Sakurai, Y Takahashi, O Yoshie, Saiki, I, CANCER RESEARCH, 66, 7, 3957, 3957,   2006 04
  • CCL17 transgenic mice show enhanced Th2-type response to both allergic and non-allergic stimuli, Y. Tsunemi, H. Saeki, K. Nakamura, D. Nagakubo, T. Nakayama, O. Yoshie, S. Kagami, K. Shimazu, T. Kadono, M. Sugaya, M. Komine, K. Matsushima, K. Tamaki, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 126, 109, 109,   2006 04
  • Enhanced anti-tumor responses induced by the combination of a couple of stimulators: IL-12 and CCL27, T Sugita, JQ Gao, N Kanagawa, Y Motomura, T Nakayama, O Yoshie, Y Hatanaka, Y Tani, H Mizuguchi, Y Tsutsumi, S Nakagawa, JOURNAL OF GENE MEDICINE, 8, 3, 397, 397,   2006 03
  • Role of fractalkine/CX3CL1 in trafficking of circulating cytotoxic effector lymphocytes that are defined by CX3CR1 expression., M Nishimura, H Umehara, T Nakayama, O Yoneda, K Hieshima, M Kakizaki, N Dohmae, O Yoshie, T Imai, FASEB JOURNAL, 16, 5, A1212, A1212,   2002 03
  • Human megakaryoblastic cell line, MEG-01, produces thymus and activation-regulated chemokine (TARC): Possible involvement of platelets in allergic inflammation, H Nishimori, T Fujisawa, Y Kato, Y Komadu, H Kamiya, T Nakayama, O Yoshie, JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 109, 1, S64, S64,   2002 01
  • Proinflammatory cytokines induce liver and activation-regulated chemokine/macrophage inflammatory protein-3 alpha/CCL20 in mucosal epithelial cells through NF-kappaB (vol 13, pg 1255, 2001), S Fujiie, K Hieshima, D Izawa, T Nakayama, R Fujisawa, H Ohyanagi, O Yoshie, INTERNATIONAL IMMUNOLOGY, 13, 11, U2, U2,   2001 11

Research Grants & Projects

  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Memory CTL-inducing vaccine sistem targeting P2 recepors
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Establishment of skin inflammation drug to accumulate high-functional regulatory T cells by single-cell gene expression analysis, We identified several regulatory T cell (Treg) subsets in inflamed skin tissue of mouse line expressing photoconvertible protein KikGR, by which we can distinguish skin-coming and -remaining lymphocytes, by multi-parameter single-cell gene and protein expression analysis. Each subset expressed each set of functional and migration-related molecules and showed different capabilities to remain in inflamed skin. We distinguished skin-coming and -remaining Tregs without KikGR expression data by multi-parameter single-cell expression analysis of functional and migration-related molecules. We also tried to identify master molecules of Treg subset differentiation and establish method to collect the Treg subsets. This research suggests that Tregs expressing different functional molecules showed different migration status in inflamed tissues.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Role of CCL28 for mucosal immunity, CCL28 is expressed in the mucosal tissues and to attract IgA-antibody secreting cells (IgA-ASCs) via CCR10. CCL28 has an antimicrobial activity against microbes in vitro. However, in vivo evidence for the role of CCL28 in the mucosal immunity remains scanty. We generated CCL28-deficient mice and demonstrated that CCL28-deficient mice showed reduced numbers and altered distribution of IgA-ASCs in the colon. The IgA contents in the fecal extracts were low. The average amounts of IgA secreted by a single IgA-ASC isolated from the lamina propria of the colon was reduced. Furthermore, the 16S rRNA sequencing analysis of feces revealed an increase of the Class Bacilli. Consistent with the low IgA production and altered microbiota in the colon, CCL28-deficient mice had aggravated colitis upon treatment with dextran sulfate, which was ameliorated by oral antibiotics. Therefore, CCL28 has an role in the mucosal immunity of the colon as a chemoattractant with a direct antimicrobial activity.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Screening of natural medicines with chemokine receptor antagonists for allergy therapy, Chemokine receptors CCR3 and CCR4 have been paid attention as potent therapeutic targets for allergic diseases. First, we screened 80 crude drugs/herbs to identify candidate substances using chemotaxis assay. Ephedra Herb inhibited the chemotaxis mediated by both CCR3 and CCR4. Thus, we examined inhibitory effects of ephedrine, a major component of Ephedra Herb. However, ephedrine exhibited little effects on the chemotaxis mediated by CCR3, CCR4, and CCR8. Therefore, we fractionated Ephedra Herb into four subfractions and examined the inhibitory effects of each subfraction. Ethyl acetate-insoluble fraction exhibited the inhibitory effects on chemotaxis and calcium mobilization mediated by CCR3 and CCR4 most significantly. Taken together, our data suggest that these crude drugs/herbs might be useful sources to develop new drugs targeting Th2-mediated allergic diseases.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Transcriptional regulation of CCR7 expression in mature T-cell malignancies, Previously, we have examined the transcriptional regulation of CCR4 expression in adult T-cell leukemia/lymphoma (ATL) and demonstrated that an AP-1 family member FRA-2 is aberrantly expressed in ATL and in association with JUND promotes CCR4 expression and cell growth together with upregulation of various proto-oncogenes such as c-Myb and SOX4 in ATL. In the present study, we have extended our analysis to CCR7, which is highly expressed in ATL and cutaneous T cell lymphomas (CTCLs), and found a novel DNA-binding element GAGGAG in the CCR7 promoter and binding of ZFYVE19 to this element. Our study suggests an existence of another oncogenic cascade in ATL and CTCLs, providing useful information on the diagnosis and treatment of mature T cell malignancies.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), The role of Fra-2 and SOX4 oncogene cascade in ATL and CTCLs, Previously, we have shown that Fra-2 is consistently expressed and involved in CCR4 expression as well as cell growth in CCR4+ mature T-cell leukemias/lymphomas, including adult T-cell leukemia/lymphoma (ATL) and cutaneous T-cell lymphomas (CTCLs). Here, we we examine the expression and function of SOX4 in the oncogenesis of ATL and CTCLs. Fra-2 and SOX4 were consistently co-expressed in the clinical samples of ATL and CTCL. SOX4 promoter analysis demonstrated that Fra-2-JunD directly activates the SOX4 promoter via an AP-1 site. Furthermore, SOX4 siRNA significantly suppressed cell growth of ATL and CTCL cell lines. We found that SOX4 knockdown reduced the expression of genes such as GCKR, NAP1 and HDAC8. We also showed direct activation of the HDAC8 promoter by SOX4. Furthermore, HDAC8 knockdown significantly suppressed cell growth of ATL and CTCL cell lines. Taken together, these finding suggest that the Fra-2-SOX4 pathway has an important oncogenic role in ATL and CTCLs.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), The role of c-Myb and ABCG2/BCRP in the drug resistance of mature T-cell lymphomas, Previously, we have shown that an AP-1 family member, FRA-2, is consistently expressed at high levels in Adult T-cell leukemia/lymphoma (ATLL) and up-regulates the expression of CCR4 as well as that of several proto-oncogenes such as c-Myb, MDM2, and BCL6. Here, we identified an ABC transporter, ABCG2/BCRP, as a target gene of c-Myb in mature T-cell lymphomas including ATLL and cutaneous T-cell lymphomas (CTCLs). We detected the transcripts and proteins of c-Myb and ABCG2/BCRP in ATLL cells. Furthermore, Ko143, a selective ABCG2/BCRP inhibitor, enhanced the antitumor activity of doxorubicin against ATLL cell lines. Taken together, our findings support the notion that the c-Myb-ABCG2/BCRP pathway plays a role in chemoresistance of mature T-cell lymphomas.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), The role of Fra-2 and c-Myb in the oncogenesis of cutaneous T-cell lymphomas, In this study, we showed that Fra-2 and c-Myb were consistently co-expressed in cutaneous T-cell lymphomas (CTCLs), and contributed to cell proliferation. Furthermore, we showed that c-Myb was a direct target gene of Fra-2 in CTCLs. In addition, we identified that RASGRP2 was a c-Myb target gene and contributed to cell proliferation in CTCLs. Taken together, our findings support the notion that the Fra-2-c-Myb oncogenic pathway plays a major role in the oncogenesis of CTCLs.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Study on the pathophysiological role of the Th2-type chemokine receptor CCR4, CCR4 is a chemokine receptor selectively expressed by Th2 cells, regulatory T cells, and skin-homing T cells. We have shown a potent ther apeutic effect of a small molecule CCR4 inhibitor on Th2-induced airway inflammation in mice. We have also shown that CCR4 and its ligand MDC/CCL22 mediate immunosuppressive interactions of dendritic cells and regulatory T cells in mesenteric lymphonodes. Furthermore, human oncogenic viruses such as EBV and HTLV-1 commonly induce expression of MDC/CCL22 in infected host cells to attract Th2 cells and regulatory T cells for immunoevasion and virus propagation.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Identification of eotaxin-3/CCL26 as a novel functional ligand for CX3CR1
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Transcriptional regulation of CCR10 expression in plasma cells, CCR10 has been reported to be expressed by almost all IgA-secreting plasma cells, while its ligand CCL28 is widely expressed in various mucosal tissues. Thus, the CCR10-CCL28 system is likely to contribute a wide distribution of IgA-secreting plasma cells in the common mucosal immune system. Here we examined the mechanism of CCR10 expression in terminally differentiating B cells. As reported previously, activated human CD19^+ B cells treated with IL-21 efficiently differentiated into IgD^-CD38^+ plasma cells. Even though IgD^-CD38^+ cells did not spontaneously expressed CCR10, a substantial fraction turned to express CCR10 if the differentiation was induced in the presence of 1,25-dihydroxyvitamin D3 (1,25-(OH)_2D_3), which is known to promote common mucosal immune responses if used as an adjuvant. However, we observed little increases in IgA^+ cells by 1,25-(OH)_2D_3. To determine the transcriptional mechanism regulating 1,25-(OH)_2D_3-inducible expression of CCR10 in terminally differentiating B cells, we next carried out a series of analysis on the CCR10 promoter. The reporter assays involving a series of 5'-deleted promoter fragments and promoter fragments with site-directed mutations revealed that a proximal Ets-1 site and an upstream vitamin D3 response element (VDRE) were critical for CCR10 expression. The NoShift transcription factor binding assays using nuclear extracts from CCR10-expressing myeloma cell lines confirmed specific binding of Ets-1 and 1,25-(OH)_2D_3-activated vitamin D3 receptor (VDR) to the respective elements. Collectively, 1,25-(OH)2D3 efficiently induces CCR10 expression in IL-21-induced human plasma cells in vitro. Furthermore, the CCR10 promoter is cooperatively activated by 1,25-(OH)_2D_3-activated VDR and Ets-1.
  • Grant-in-Aid for Scientific Research, Study on Chemokine