詳細

宮澤　正顯 （ミヤザワ　マサアキ）

  • 医学科　客員教授
Last Updated :2023/11/29

コミュニケーション情報 byコメンテータガイド

  • コメント

    エイズウイルスやインフルエンザウイルスに対する宿主の抵抗性を決める遺伝子を研究しています。厚生労働科研費の研究代表者を長く務め、イタリア･フランス･タイなどと共同研究しています。

  • 報道関連出演・掲載一覧

    ＜報道関連出演・掲載一覧＞ ●2021/12/16 　読売テレビ「かんさい情報ネットten.」 　3回目のワクチン接種について ●2021/11/30 　関西テレビ「報道ランナー」 　オミクロン株について ●2021/11/9 　関西テレビ「報道ランナー」 　ワクチン接種後の抗体量が減少した場合の影響について ●2021/10/29 　関西テレビ「報道ランナー 　ワクチン3回目接種について ●2021/9/10 　読売テレビ「かんさい情報ネットten.」 　ノババックス製の新型コロナワクチンについて ●2021/8/27 　読売テレビ「かんさい情報ネットten.」 　学校、職場、家庭内で感染を防ぐために必要なことについて ●2021/8/23 　関西テレビ「報道ランナー」 　ワクチンについて ●2021/8/10 　関西テレビ「報道ランナー」 　ワクチンについて ●2021/6/25 　読売テレビ「かんさい情報ネットten.」 　新型コロナのインド型変異ウイルス「デルタ株」について ●2021/6/3 　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルスの変異株について ●2021/5/28 　読売テレビ「かんさい情報ネットten」 　新型コロナウイルスのインド株について ●2021/5/20 　読売テレビ「かんさい情報ネットten.」 　ワクチン承認について ●2021/5/9 　フジテレビ「Mr.サンデー」 　新型コロナウイルスの変異ウイルスについて ●2021/5/7 　読売テレビ「かんさい情報ネットten.」 　オリンピック選手へのファイザー製ワクチン提供ついて ●2021/4/2 　読売テレビ「かんさい情報ネットten」 　新型コロナ第4波について ●2021/1/26 　読売テレビ「かんさい情報ネットten.」 　コロナウイルスのワクチン接種の痛みについて ●2021/1/14 　読売テレビ「情報ライブミヤネ屋」 　新型コロナウイルスについて ●2020/10/16 　読売テレビ「かんさい情報ネットten.」 　コロナウイルスの外国での流行について ●2020/9/28 　毎日新聞 　世界の死者が100万人に迫っていることから冬にむけて感染者増加について ●2020/9/19 　産経新聞 　イベントの人数制限が緩和されることからコロナウイルス感染再拡大のリスクについて ●2020/9/11 　読売テレビ「かんさい情報ネットten.」 　WHOパンデミック宣言から半年が経つ今、現状や今後について ●2020/8/23 　朝日新聞 　夏風邪とコロナの見分け方 ●2020/8/12 　読売テレビ「情報ライブミヤネ屋」 　コロナウイルスについて ●2020/7/9 　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルス感染拡大について ●2020/6/7 　読売新聞 　コロナウイルスのワクチンについて ●2020/6/5 　NHK「かんさい熱視線」 　新型コロナウイルスについて ●2020/6/4 　読売新聞 　コロナウイルスの「交差免疫説」について ●2020/6/1 　読売テレビ「情報ライブミヤネ屋」 　コロナウイルスについて ●2020/5/26 　時事通信 　外出自粛が解除後、「第2波」への懸念について ●2020/5/25 　読売テレビ「情報ライブミヤネ屋」 　コロナウイルスによる緊急事態宣言の全面解除について ●2020/5/22 　読売新聞　産経新聞 　コロナウイルスによる緊急事態宣言の解除を受けて ●2020/5/15 　読売新聞 　コロナウイルスの抗体検査について ●2020/5/8 　夕刊フジ 　レムデシビルについて ●2020/5/7 　読売テレビ「情報ライブミヤネ屋」 　コロナウイルスの抗体検査について ●2020/5/6 　読売テレビ「情報ライブミヤネ屋」 　コロナウイルスについて ●2020/5/2 　毎日新聞 　大阪府のコロナウイルス感染者数推移について ●2020/4/24　 　NHK「かんさい熱視線」 　新型コロナウイルス感染症について ●2020/4/23　 　読売テレビ「情報ライブ ミヤネ屋」 　新型コロナウイルス感染症について ●2020/4/22　 　産経新聞 　新型コロナウイルス感染症について ●2020/4/21 　毎日放送「ミント！」「Newsミント！」 　新型コロナウイルス感染症について ●2020/4/20　 　スポーツ報知 　新型コロナウイルス感染症について ●2020/4/16　 　読売新聞　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルス感染症について ●2020/4/15　 　朝日新聞 　新型コロナウイルス感染症について ●2020/4/10　 　関西テレビ「報道ランナー」 　新型コロナウイルス感染症について ●2020/4/9　 　産経新聞　毎日放送「ミント！」 　新型コロナウイルス感染症について ●2020/4/8　 　信濃毎日新聞　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルス感染症について ●2020/4/7　 　サンケイスポーツ　関西テレビ「報道ランナー」　岩手めんこいテレビ「mit Live News」 　新型コロナウイルス感染症について ●2020/4/6 　朝日放送ラジオ「おはようパーソナリティ道上洋三です」 　コロナウイルスについて電話出演 ●2020/4/4　 　産経新聞 　新型コロナウイルス感染症について ●2020/4/2　 　毎日放送「ちちんぷいぷい」　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルス感染症について ●2020/4/1　 　毎日放送「ミント！」 　新型コロナウイルス感染症について ●2020/3/20　 　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルス感染症について ●2020/3/16 　毎日放送「ミント！」 　新型コロナウイルス感染症について ●2020/3/13　 　読売テレビ「かんさい情報ネットten.」 　新型コロナウイルス感染症について ●2020/3/5　 　毎日放送「ちちんぷいぷい」 　新型コロナウイルス感染症について ●2020/3/4　 　毎日放送「ミント！」 　新型コロナウイルス感染症について ●2020/2/20　 　毎日放送「ちちんぷいぷい」 　新型コロナウイルス感染症について ●2020/2/18　 　毎日放送「ミント！」 　新型コロナウイルス感染症について ●2020/2/14　 　毎日放送「ちちんぷいぷい」 　新型コロナウイルス感染症について ●2020/2/10　 　読売テレビ「情報ライブ ミヤネ屋」 　新型コロナウイルス感染症について ●2020/2/3　 　読売テレビ「情報ライブ ミヤネ屋」 　新型コロナウイルス感染症について ●2020/1/30　 　夕刊フジ 　新型コロナウイルス感染症について ●2019/4/16 　朝日放送「キャスト」 　リウマチとがんの関係について ●2019/1/14 　朝日新聞 　ウィルス遺伝子について●2015/11/17 　日本テレビ系列「情報ライブ ミヤネ屋」 　HIVの治療法について ●2015/6/25 　朝日放送「キャスト」 　ＨＩＶ感染に関する医学的根拠について ●2014/8/13 　毎日放送「ちちんぷいぷい」 　エボラ出血熱について ●2014/8/5 　毎日放送「ちちんぷいぷい」 　STAP細胞について"

研究者情報

学位

  • 医学博士（1987年09月 東北大学）
  • 医学士（1982年03月 東北大学）

ホームページURL

J-Global ID

プロフィール

  • 東北大学医学部1982年卒業、医師、医学博士(東北大学)
    東北大学助手、米国NIH/NIAID Visiting Associate、三重大学医学部助教授を経て、1996年近畿大学医学部教授(免疫学教室担当)
    近畿大学大学院医学研究科長、近畿大学医学部共同研究施設長、近畿大学遺伝子組換え実験安全主任者・バイオセーフティ委員長を歴任


    2023年3月　近畿大学定年退職、4月　近畿大学名誉教授


    2023年4月より、鹿児島市に本店を置く（株）新日本科学で経鼻粘膜ワクチン研究開発センター長

研究キーワード

  • エピトープ   CD4陽性T細胞   Tリンパ球   ワクチン   効果的なワクチンの開発   自己免疫病   免疫応答遺伝子   感染抵抗性   Autoimmune disease   Immune regulatory genes   Resistance to infection   

現在の研究分野（キーワード）

    エイズウイルスやインフルエンザウイルスに対する宿主の抵抗性を決める遺伝子を研究しています。厚生労働科研費の研究代表者を長く務め、イタリア･フランス･タイなどと共同研究しています。

研究分野

  • ライフサイエンス / 免疫学
  • ライフサイエンス / ウイルス学
  • ライフサイエンス / 実験病理学
  • ライフサイエンス / 医化学

経歴

  • 2023年04月 - 現在  近畿大学医学部客員教授
  • 2023年04月 - 現在  株式会社新日本科学経鼻粘膜ワクチン研究開発センターセンター長・理事
  • 1996年04月 - 2023年03月  近畿大学医学部免疫学教室主任教授
  • 2016年10月 - 2022年09月  近畿大学大学院医学研究科研究科長
  • 1995年08月 - 1996年03月  Associate Professor, Mie University School of Medicine
  • 1995年 - 1996年  三重大学医学部 助教授Faculty of Medicine
  • 1984年01月 - 1995年07月  Research Associate, Tohoku University School of Medicine
  • 1984年 - 1995年  東北大学医学部 助手Faculty of Medicine
  • 1986年09月 - 1990年01月  Visiting Associate, NIH, NIAID, U.S.A.
  • 1986年 - 1990年  アメリカ合衆国国立感染症アレルギー研究所客員研究員

学歴

  •         - 1983年   東北大学   医学研究科   病理学系専攻
  •         - 1983年   東北大学   Division of Medical Science
  •         - 1982年   東北大学   医学部   医学科
  •         - 1982年   東北大学   Faculty of Medicine

所属学協会

  • 日本ワクチン学会   アメリカ微生物学会(American Society for Microbiology)   日本生体防御学会   日本癌学会   日本免疫学会   日本病理学会   American Society for Microbiology   

研究活動情報

論文

  • Yoshiyuki Hakata; Kazuma Yamashita; Sonoko Hashimoto; Takashi Ohtsuki; Masaaki Miyazawa; Mizuki Kitamatsu
    Pharmaceutics 15 4 2023年03月 
    A connection of a functional peptide with a cell-penetrating peptide (CPP) used a heterodimeric coiled-coil as a molecular zipper can improve the intracellular delivery and activity of the functional peptide. However, the chain length of the coiled coil required for functioning as the molecular zipper is unknown at present. To solve the problem, we prepared an autophagy-inducing peptide (AIP) that conjugates with the CPP via heterodimeric coiled-coils consisting of 1 to 4 repeating units (K/E zipper; AIP-Kn and En-CPP), and we investigated the optimum length of the K/E zipper for effective intracellular delivery and autophagy induction. Fluorescence spectroscopy showed that K/E zippers with n = 3 and 4 formed a stable 1:1 hybrid (AIP-K3/E3-CPP and AIP-K4/E4-CPP, respectively). Both AIP-K3 and AIP-K4 were successfully delivered into cells by the corresponding hybrid formation with K3-CPP and K4-CPP, respectively. Interestingly, autophagy was also induced by the K/E zippers with n = 3 and 4, more intensively by the former than by the latter. The peptides and K/E zippers used in this study did not show significant cytotoxicity. These results indicate that the effective induction of autophagy occurs via an exquisite balance of the association and dissociation of the K/E zipper in this system.
  • Tomomasa Izumiyama; Masaaki Miyazawa
    Modern Rheumatology 2022年09月 [査読有り]
     
    ABSTRACT Rheumatoid arthritis (RA) has long been characterized by synovitis and bone erosions typically developing symmetrically in small joints. However, recent advances in imaging modalities have indicated frequent association of tenosynovitis with RA, and some consider tenosynovitis to be not just a complication but a major trait of RA. Further, as there are cases with tenosynovitis preceding the clinical detection of inflammatory arthritis in predisposed individuals, tenosynovitis may constitute an important biomarker in defining the pre-RA phase of disease development. Tenosynovitis itself must be treated as it causes functional impairment and physical as well as socioeconomic burden, and its treatment may result in effective prevention of RA development at a pre-arthritic stage. Thus, further efforts need to be taken in detecting and treating tenosynovitis in pre-RA stage, which can be facilitated by ultrasonography and magnetic resonance imaging.
  • Masaaki Miyazawa; Mario Clerici
    Frontiers in Virology 2 2022年06月 [査読有り][招待有り]
  • Kenta Teruya; Ayumi Oguma; Satoko Takahashi; Miki Watanabe-Matsui; Sachiyo Tsuji-Kawahara; Masaaki Miyazawa; Katsumi Doh-ura
    International Immunopharmacology 107 108672 - 108672 2022年06月 [査読有り]
     
    The anti-prion activity of cellulose ether (CE) has been reported in rodents, but the mechanism of action is not well understood. As defects in early T-cell development have been reported in Tga20 mice which show only a slight effect of CE administration, we investigated the involvement of immune functions in the CE action. We confirmed an insertion of the prion protein transgene into the pre T-cell antigen receptor α gene of Tga20 mice, and its impaired expression in the thymus and other tissues. The influence of immune suppression on the CE effect was then examined in high CE-responder mice treated with immunosuppressive agents or neonatal thymectomy. As neonatal thymectomy significantly reduced the CE effect, we compared the influence of various T-cell defects in mice with similar genetic backgrounds. The CE effect was increased or unchanged in mice with defects in the αβ T-cell lineage, whereas it was abolished in T-cell receptor δ deficient mice. Further, when other immune defects were examined, the CE effect was reduced in mice with lysosomal trafficking dysfunction, but was unchanged in mice deficient in B-cell differentiation or toll-like receptor 4 signaling. These findings collectively suggest that the mechanism of CE action may involve γδ T cells and lytic granule function, as well as immune factors like natural killer T cells which are lacking in pre T-cell antigen receptor α deficient mice and neonatally thymectomized mice.
  • Shota Tsukimoto; Yoshiyuki Hakata; Sachiyo Tsuji-Kawahara; Takuji Enya; Tetsuo Tsukamoto; Seiya Mizuno; Satoru Takahashi; Shinichi Nakao; Masaaki Miyazawa
    Viruses 14 4 832 - 832 2022年04月 [査読有り]
     
    Tissue and subcellular localization and its changes upon cell activation of virus-restricting APOBEC3 at protein levels are important to understanding physiological functions of this cytidine deaminase, but have not been thoroughly analyzed in vivo. To precisely follow the possible activation-induced changes in expression levels of APOBEC3 protein in different mouse tissues and cell populations, genome editing was utilized to establish knock-in mice that express APOBEC3 protein with an in-frame FLAG tag. Flow cytometry and immunohistochemical analyses were performed prior to and after an immunological stimulation. Cultured B cells expressed higher levels of APOBEC3 protein than T cells. All differentiation and activation stages of freshly prepared B cells expressed significant levels of APOBEC3 protein, but germinal center cells possessed the highest levels of APOBEC3 protein localized in their cytoplasm. Upon immunological stimulation with sheep red blood cells in vivo, germinal center cells with high levels of APOBEC3 protein expression increased in their number, but FLAG-specific fluorescence intensity in each cell did not change. T cells, even those in germinal centers, did not express significant levels of APOBEC3 protein. Thus, mouse APOBEC3 protein is expressed at distinctively high levels in germinal center B cells. Antigenic stimulation did not affect expression levels of cellular APOBEC3 protein despite increased numbers of germinal center cells.
  • Shunichi Shiozawa; Ken Tsumiyama; Yumi Miyazaki; Kenichi Uto; Keiichi Sakurai; Toshie Nakashima; Hiroko Matsuyama; Ai Doi; Miho Tarui; Manabu Izumikawa; Mai Kimura; Yuko Fujita; Chisako Satonaka; Takahiko Horiuchi; Tsukasa Matsubara; Motohiro Oribe; Takashi Yamane; Hidetoshi Kagawa; Quan-Zhen Li; Keiko Mizuno; Yohei Mukai; Kazuhiro Murakami; Takuji Enya; Shota Tsukimoto; Yoshiyuki Hakata; Masaaki Miyazawa; Kazuko Shiozawa
    iScience 25 1 103537 - 103537 2022年01月 [査読有り]
     
    Pathogens including autoantigens all failed to induce systemic lupus erythematosus (SLE). We, instead, studied the integrity of host's immune response that recognized pathogen. By stimulating TCR with an antigen repeatedly to levels that surpass host's steady-state response, self-organized criticality, SLE was induced in mice normally not prone to autoimmunity, wherein T follicular helper (Tfh) cells expressing the guanine nucleotide exchange factor DOCK8 on the cell surface were newly generated. DOCK8+Tfh cells passed through TCR re-revision and induced varieties of autoantibody and lupus lesions. They existed in splenic red pulp and peripheral blood of active lupus patients, which subsequently declined after therapy. Autoantibodies and disease were healed by anti-DOCK8 antibody in the mice including SLE-model (NZBxNZW) F1 mice. Thus, DOCK8+Tfh cells generated after repeated TCR stimulation by immunogenic form of pathogen, either exogenous or endogenous, in combination with HLA to levels that surpass system's self-organized criticality, cause SLE.
  • 新規に生成されたDOCK8発現濾胞性ヘルパーT細胞が全身性エリテマトーデスを引き起こす(Newly generated DOCK8-expressing T follicular helper cells cause systemic lupus erythematosus)
    Shiozawa Shunichi; Tsumiyama Ken; Sakurai Keiichi; Matsubara Tsukasa; Yamane Takashi; Miyazawa Masaaki
    日本免疫学会総会・学術集会記録 50 Proceedings 2 - P 2021年11月
  • Newly generated DOCK8-expressing T follicular helper cells cause systemic lupus erythematosus(和訳中)
    Shiozawa Shunichi; Tsumiyama Ken; Sakurai Keiichi; Matsubara Tsukasa; Yamane Takashi; Miyazawa Masaaki
    日本免疫学会総会・学術集会記録 50 Proceedings 2 - P 2021年11月
  • 近交系卵巣明細胞癌マウスモデルの樹立と腫瘍免疫の解析
    村上 幸祐; 宮川 知保; 加藤 茂樹; 高村 史記; 宮澤 正顯; 松村 謙臣
    日本婦人科腫瘍学会学術講演会プログラム・抄録集 63回 199 - 199 (公社)日本婦人科腫瘍学会 2021年07月
  • Yoshiyuki Hakata; Masaaki Miyazawa
    Microorganisms 8 12 1976 - 1976 2020年12月 [査読有り]
     
    Apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3 (APOBEC3) proteins (APOBEC3s) are deaminases that convert cytosines to uracils predominantly on a single-stranded DNA, and function as intrinsic restriction factors in the innate immune system to suppress replication of viruses (including retroviruses) and movement of retrotransposons. Enzymatic activity is supposed to be essential for the APOBEC3 antiviral function. However, it is not the only way that APOBEC3s exert their biological function. Since the discovery of human APOBEC3G as a restriction factor for HIV-1, the deaminase-independent mode of action has been observed. At present, it is apparent that both the deaminase-dependent and -independent pathways are tightly involved not only in combating viruses but also in human tumorigenesis. Although the deaminase-dependent pathway has been extensively characterized so far, understanding of the deaminase-independent pathway remains immature. Here, we review existing knowledge regarding the deaminase-independent antiretroviral functions of APOBEC3s and their molecular mechanisms. We also discuss the possible unidentified molecular mechanism for the deaminase-independent antiretroviral function mediated by mouse APOBEC3.
  • Masaaki Miyazawa
    Inflammation and Regeneration 40 1 39  2020年10月 [査読有り][招待有り]
     
    Abstract Factors determining the progression of frequently mild or asymptomatic severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection into life-threatening pneumonia remain poorly understood. Viral and host factors involved in the development of diffuse alveolar damage have been extensively studied in influenza virus infection. Influenza is a self-limited upper respiratory tract infection that causes acute and severe systemic symptoms and its spread to the lungs is limited by CD4+ T-cell responses. A vicious cycle of CCL2- and CXCL2-mediated inflammatory monocyte and neutrophil infiltration and activation and resultant massive production of effector molecules including tumor necrosis factor (TNF)-α, nitric oxide, and TNF-related apoptosis-inducing ligand are involved in the pathogenesis of progressive tissue injury. SARS-CoV-2 directly infects alveolar epithelial cells and macrophages and induces foci of pulmonary lesions even in asymptomatic individuals. Mechanisms of tissue injury in SARS-CoV-2-induced pneumonia share some aspects with influenza virus infection, but IL-1β seems to play more important roles along with CCL2 and impaired type I interferon signaling might be associated with delayed virus clearance and disease severity. Further, data indicate that preexisting memory CD8+ T cells may play important roles in limiting viral spread in the lungs and prevent progression from mild to severe or critical pneumonia. However, it is also possible that T-cell responses are involved in alveolar interstitial inflammation and perhaps endothelial cell injury, the latter of which is characteristic of SARS-CoV-2-induced pathology.
  • Haruo Ohtani; Yoshihiro Nozaki; Takashi Murakami; Lisheng Lin; Junko Shiono; Masaaki Miyazawa
    Journal of Clinical and Experimental Hematopathology 60 3 108 - 112 2020年09月 [査読有り]
     
    We report an autopsy case of acute myocarditis, in which the mediastinal lymph nodes exhibited unique findings. A 15-year-old Japanese boy was diagnosed with the secondary onset of acute myocarditis. No viruses were identified. Autopsy confirmed acute lymphocytic myocarditis. Lymphadenopathy was observed, especially in pulmonary hilar/mediastinal areas. Microscopically, interfollicular areas were uniformly filled with medium-sized, round cells that resembled lymphocytes. They were immunohistochemically CD3- CD5- CD19+ CD20- CD79a- Pax-5- CD138+ MUM1+ LMP1- EBNA2- cytoplasmic IgG+ IgA- and IgM-. No monotypia was observed for kappa and lambda light chains, and multiplex polymerase chain reaction analyses of immunoglobulin heavy chain variable region diversity demonstrated oligoclonal peaks, suggesting reactive change. IgG+ or VS38c+ cells frequently co-expressed Ki-67 (up to 80%). We considered these cells abundantly present in lymph nodes to be reactive plasmablasts because they were early plasma cells with proliferative activity.
  • Miguel Meira e Cruz; Masaaki Miyazawa; David Gozal
    European Respiratory Journal 55 6 2001023 - 2001023 2020年06月 [査読有り]
  • Yoshiyuki Hakata; Suzuka Ishikawa; Takashi Ohtsuki; Masaaki Miyazawa; Mizuki Kitamatsu
    ORGANIC & BIOMOLECULAR CHEMISTRY 18 10 1978 - 1986 2020年03月 [査読有り]
     
    Development of an intracellular delivery method for functional peptides via cell-penetrating peptides (CPPs) expands peptide use in basic research and therapeutic applications. Although direct conjugation of a functional peptide with a CPP is the simplest method for delivery, this method has not always been reliable. CPPs usually contain several positively charged amino acids that potentially interact non-specifically with negatively charged molecules in cells and subsequently interfere with conjugated functional peptide function. Here we demonstrate a new intracellular delivery method for peptides in which a functional peptide is released from a positively charged CPP via peptide nucleic acids (PNAs). We prepared an 8-mer PNA conjugated to octa-arginine in tandem (PNA1-CPP) and linked its complementary PNA to an autophagy inducing peptide (PNA2-AIP) by solid-phase peptide synthesis. PNA1-CPP and PNA2-AIP formed a 1 : 1 hybrid via PNA1/PNA2 interaction, thereby indirectly but stably connecting the AIP to the CPP. PNA2-AIP was successfully delivered into cells in a hybrid formation-dependent manner and at least some portion of the PNA1-CPP/PNA2-AIP hybrids dissociated into PNA2-AIP and PNA1-CPP inside the cells. Notably, PNA2-AIP delivered to cells induced more autophagy than AIP directly conjugated to CPP (CPP-AIP). Further, the PNA hybrid did not induce significant cell death. These findings indicate that the PNA1/PNA2 hybrid can function as a molecular glue enabling the delivery of functional peptides into cells.
  • Koji Haratani; Kimio Yonesaka; Shiki Takamura; Osamu Maenishi; Ryoji Kato; Naoki Takegawa; Hisato Kawakami; Kaoru Tanaka; Hidetoshi Hayashi; Masayuki Takeda; Naoyuki Maeda; Takashi Kagari; Kenji Hirotani; Junji Tsurutani; Kazuto Nishio; Katsumi Doi; Masaaki Miyazawa; Kazuhiko Nakagawa
    The Journal of clinical investigation 130 1 374 - 388 2020年01月 [査読有り]
     
    Immunotherapy targeting programmed cell death-1 (PD-1) induces durable antitumor efficacy in many types of cancer. However, such clinical benefit is limited because of the insufficient reinvigoration of antitumor immunity with the drug alone; therefore, rational therapeutic combinations are required to improve its efficacy. In our preclinical study, we evaluated the antitumor effect of U3-1402, a human epidermal growth factor receptor 3-targeting (HER3-targeting) antibody-drug conjugate, and its potential synergism with PD-1 inhibition. Using a syngeneic mouse tumor model that is refractory to anti-PD-1 therapy, we found that treatment with U3-1402 exhibited an obvious antitumor effect via direct lysis of tumor cells. Disruption of tumor cells by U3-1402 enhanced the infiltration of innate and adaptive immune cells. Chemotherapy with exatecan derivative (Dxd, the drug payload of U3-1402) revealed that the enhanced antitumor immunity produced by U3-1402 was associated with the induction of alarmins, including high-mobility group box-1 (HMGB-1), via tumor-specific cytotoxicity. Notably, U3-1402 significantly sensitized the tumor to PD-1 blockade, as a combination of U3-1402 and the PD-1 inhibitor significantly enhanced antitumor immunity. Further, clinical analyses indicated that tumor-specific HER3 expression was frequently observed in patients with PD-1 inhibitor-resistant solid tumors. Overall, U3-1402 is a promising candidate as a partner of immunotherapy for such patients.
  • Shiki Takamura; Shigeki Kato; Chihiro Motozono; Takeshi Shimaoka; Satoshi Ueha; Kazuhiko Matsuo; Kosuke Miyauchi; Tomoko Masumoto; Asami Katsushima; Takashi Nakayama; Michio Tomura; Kouji Matsushima; Masato Kubo; Masaaki Miyazawa
    The Journal of experimental medicine 216 12 2736 - 2747 2019年12月 [査読有り]
     
    Populations of CD8+ lung-resident memory T (TRM) cells persist in the interstitium and epithelium (airways) following recovery from respiratory virus infections. While it is clear that CD8+ TRM cells in the airways are dynamically maintained via the continuous recruitment of new cells, there is a vigorous debate about whether tissue-circulating effector memory T (TEM) cells are the source of these newly recruited cells. Here we definitively demonstrate that CD8+ TRM cells in the lung airways are not derived from TEM cells in the circulation, but are seeded continuously by TRM cells from the lung interstitium. This process is driven by CXCR6 that is expressed uniquely on TRM cells but not TEM cells. We further demonstrate that the lung interstitium CD8+ TRM cell population is also maintained independently of TEM cells via a homeostatic proliferation mechanism. Taken together, these data show that lung memory CD8+ TRM cells in the lung interstitium and airways are compartmentally separated from TEM cells and clarify the mechanisms underlying their maintenance.
  • Yoshiyuki Hakata; Jun Li; Takahiro Fujino; Yuki Tanaka; Rie Shimizu; Masaaki Miyazawa
    PLOS Pathogens 15 12 e1008173  2019年12月 [査読有り]
     
    Mouse APOBEC3 (mA3) inhibits murine leukemia virus (MuLV) replication by a deamination-independent mechanism in which the reverse transcription is considered the main target process. However, other steps in virus replication that can be targeted by mA3 have not been examined. We have investigated the possible effect of mA3 on MuLV protease-mediated processes and found that mA3 binds both mature viral protease and Pr180gag-pol precursor polyprotein. Using replication-competent MuLVs, we also show that mA3 inhibits the processing of Pr65 Gag precursor. Furthermore, we demonstrate that the autoprocessing of Pr180gag-pol is impeded by mA3, resulting in reduced production of mature viral protease. This reduction appears to link with the above inefficient Pr65gag processing in the presence of mA3. Two major isoforms of mA3, exon 5-containing and -lacking ones, equally exhibit this antiviral activity. Importantly, physiologically expressed levels of mA3 impedes both Pr180gag-pol autocatalysis and Pr65gag processing. This blockade is independent of the deaminase activity and requires the C-terminal region of mA3. These results suggest that the above impairment of Pr180gag-pol autoprocessing may significantly contribute to the deaminase-independent antiretroviral activity exerted by mA3.
  • 原谷 浩司; 米阪 仁雄; 高村 史記; 前西 修; 宮澤 正顯; 中川 和彦
    肺癌 59 6 568 - 568 (NPO)日本肺癌学会 2019年11月
  • 塚本徹雄; 河原佐智代; 宮澤正顯
    Bio Clinica 34 6 629 - 635 株式会社 北隆館 2019年06月 [査読有り][招待有り]
     
    マウスのフレンドウイルス(FV)感染による赤白血病発症機構については，膜結合型チロシンキナーゼSf-Stk依存性多段階発がん説が主流であったが，我々グループはSf-Stkをもたないマウスでの発症を実証した。その一方で，Sf-Stk非依存性発症機構としてストレス造血が注目されているが，FV感染後ストレス造血が惹起される分子機構は全く不明である。ここではFV誘発赤白血病についてこれまでの知見を総括し，白血病研究におけるFV感染マウスモデルの位置付けを概観しつつ，さらなる研究の意義について考察する。
  • Yoshiyuki Hakata; Hiroyuki Michiue; Takashi Ohtsuki; Masaaki Miyazawa; Mizuki Kitamatsu
    Bioorganic & medicinal chemistry letters 29 7 878 - 881 2019年04月 [査読有り]
     
    We synthesized a pair of compounds containing leucine zipper peptides to deliver protein cargo into cells. One is a cell-penetrating peptide (CPP) with Lz(E), a leucine zipper peptide containing negatively charged amino acids, and the other is a Nanog protein with Lz(K), a leucine zipper peptide containing positively charged amino acids. When cells were treated with these equimolar mixtures, Nanog-Lz(K) hybridized with Lz(E)-CPP was successfully delivered into the cells. Furthermore, Nanog-Lz(K) exerted its proper function after nuclear transport.
  • Meira e Cruz, M; M. Miyazawa; R. Manfredini; D. Cardinali; J. A. Madrid; R. Reiter; J. F. Araujo; R. Agostinho; D. Acuña-Castroviejo
    Eur. J. Intern. Med. 60 1 - 3 2019年02月 [査読有り]
  • 全身性自己免疫疾患 全身性エリテマトーデス(SLE)を引き起こす自己抗体産生誘導性CD4 T(aiCD4 T)細胞としてのDock8陽性CD4 T細胞 SLEの原因としての自己臨界点説の概念の立証(Systemic autoimmune diseases-3 Dock8-Positive CD4 T cell as Autoantibody-Inducing CD4 T(aiCD4 T) Cell That Causes Systemic Lupus Erythematosus(SLE): Proof of Concept of Self-Organized Criticality Theory as a Cause of SLE)
    Shiozawa Shunichi; Tsumiyama Ken; Miyazaki Yumi; Sakurai Keiichi; Miyazawa Masaaki
    日本免疫学会総会・学術集会記録 47 Proceedings 2 - P 2018年12月
  • Shiozawa Shunichi; Tsumiyama Ken; Miyazaki Yumi; Sakurai Keiichi; Miyazawa Masaaki
    ARTHRITIS & RHEUMATOLOGY 70 2018年09月 [査読有り]
  • Kimio Yonesaka; Koji Haratani; Shiki Takamura; Hitomi Sakai; Ryoji Kato; Naoki Takegawa; Takayuki Takahama; Kaoru Tanaka; Hidetoshi Hayashi; Masayuki Takeda; Sigeki Kato; Osamu Maenishi; Kazuko Sakai; Yasutaka Chiba; Takafumi Okabe; Keita Kudo; Yoshikazu Hasegawa; Hiroyasu Kaneda; Michiko Yamato; Kenji Hirotani; Masaaki Miyazawa; Kazuto Nishio; Kazuhiko Nakagawa
    Clinical Cancer Research 24 11 2653 - 2664 2018年06月 [査読有り]
  • Shiozawa Shunichi; Tsumiyama Ken; Miyazaki Yumi; Sakurai Keiichi; Miyazawa Masaaki
    JOURNAL OF IMMUNOLOGY 200 1 2018年05月 [査読有り]
  • Masayuki Fujino; Hirotaka Sato; Tomotaka Okamura; Akihiko Uda; Satoshi Takeda; Nursarat Ahmed; Shigeyuki Shichino; Teiichiro Shiino; Yohei Saito; Satoru Watanabe; Chie Sugimoto; Marcelo J. Kuroda; Manabu Ato; Yoshiyuki Nagai; Shuji Izumo; Kouji Matsushima; Masaaki Miyazawa; Aftab A. Ansari; Francois Villinger; Kazuyasu Mori
    JOURNAL OF VIROLOGY 91 13 2017年07月 [査読有り]
     
    Glycosylation of Env defines pathogenic properties of simian immunodeficiency virus (SIV). We previously demonstrated that pathogenic SlVmac239 and a live-attenuated, quintuple deglycosylated Env mutant (Delta 5G) virus target CD4(+) T cells residing in different tissues during acute infection. SlVmac239 and A5G preferentially infected distinct CD4(+) T cells in secondary lymphoid organs (SLOs) and within the lamina propria of the small intestine, respectively (C. Sugimoto et al., J Virol 86: 9323-9336, 2012, https://doi.org/10.1128/JVI.00948-12). Here, we studied the host responses relevant to SIV targeting of CXCR3(+) CCR5(+) CD4(+) T cells in SLOs. Genome-wide transcriptome analyses revealed that Th1-polarized inflammatory responses, defined by expression of CXCR3 chemokines, were distinctly induced in the SlVmac239-infected animals. Consistent with robust expression of CXCL10, CXCR3(+) T cells were depleted from blood in the SlVmac239-infected animals. We also discovered that elevation of CXCL10 expression in blood and SLOs was secondary to the induction of CD14(+) CD16(+) monocytes and MAC387(+) macrophages, respectively. Since the significantly higher levels of SIV infection in SLOs occurred with a massive accumulation of infiltrated MAC387(+) macrophages, T cells, dendritic cells (DCs), and residential macrophages near high endothelial venules, the results highlight critical roles of innate/inflammatory responses in SlVmac239 infection. Restricted infection in SLOs by A5G also suggests that glycosylation of Env modulates innate/inflammatory responses elicited by cells of monocyte/macrophage/DC lineages. IMPORTANCE We previously demonstrated that a pathogenic SlVmac239 virus and a live-attenuated, deglycosylated mutant A5G virus infected distinct CD4(+) T cell subsets in SLOs and the small intestine, respectively (C. Sugimoto et al., J Virol 86:9323-9336, 2012, https://doi.org/10.1128/JVI,00948-12). Accordingly, infections with SlVmac239, but not with A5G, deplete CXCR3(+) CCR5(+) CD4(+) T (Th1) cells during the primary infection, thereby compromising the cellular immune response. Thus, we hypothesized that distinct host responses are elicited by the infections with two different viruses. We found that SlVmac239 induced distinctly higher levels of inflammatory Th1 responses than Delta 5G. In particular, SlVmac239 infection elicited robust expression of CXCL10, a chemokine for CXCR3(+) cells, in CD14(+) CD16(+) monocytes and MAC387(+) macrophages recently infiltrated in SLOs. In contrast, Delta 5G infection elicited only modest inflammatory responses. These results suggest that the glycosylation of Env modulates the inflammatory/Th1 responses through the monocyte/macrophage subsets and elicits marked differences in SIV infection and clinical outcomes.
  • Takeshi Yoshikawa; Jianfeng Wu; Motoyuki Otsuka; Takahiro Kishikawa; Nobumi Suzuki; Akemi Takata; Motoko Ohno; Rei Ishibashi; Mari Yamagami; Ryo Nakagawa; Naoya Kato; Masaaki Miyazawa; Jiahuai Han; Kazuhiko Koike
    GASTROENTEROLOGY 152 3 631 - 643 2017年02月 [査読有り]
     
    BACKGROUND & AIMS: Little is known about the mechanisms by which chronic inflammation contributes to carcinogenesis, such as the development of colon tumors in patients with inflammatory bowel diseases. Specific microRNA (miRNAs) can function as suppressors or oncogenes, and widespread alterations in miRNA expression have been associated with tumorigenesis. We studied whether alterations in miRNA function contribute to inflammation-associated colon carcinogenesis. METHODS: We studied the effects of inflammatory cytokines, such as tumor necrosis factor, interleukin-1 alpha (IL1A), and IL1 beta (IL1B), on miRNA function, measured by activity of reporter constructs containing miRNA-binding sites in their 30 untranslated regions, in human 293T embryonic kidney, Caco-2, HT29, and HCT116 colon carcinoma cells, as well as dicer(+/+) and dicer(-/-), and Apobec3(+/+) and Apobec3(-/-) mouse embryonic fibroblasts. Cells were analyzed by immunoblots, immunohistochemistry, and flow cytometry. We generated transgenic mice expressing reporter constructs regulated by LET7B, MIR122, and MIR29b response elements; some mice were given injections of miRNA inhibitors (anti-MIR122 or anti-LET7B), a negative control, or tumor necrosis factor. Liver tissues were collected and analyzed by immunoblotting. Reporter mice were given azoxymethane followed by dextran sulfate sodium to induce colitis and colon tumors; some mice were given the ROCK inhibitor fasudil along with these agents (ROCK inhibitors increase miRNA function). Colon tissues were collected and analyzed by immunohistochemistry, immunoblots, and fluorescence microscopy. RESULTS: Incubation of cell lines with inflammatory cytokines reduced the ability of miRNAs to down-regulate expression from reporter constructs; dicer was required for this effect, so these cytokines relieve miRNA-dependent reductions in expression. The cytokines promoted degradation of APOBEC3G, which normally promotes miRNA loading into argonaute 2-related complexes. Mice with colitis had reduced miRNA function, based on increased expression of reporter genes. Administration of fasudil to mice did not reduce the severity of colitis that developed but greatly reduced the numbers of colon tumors formed (mean 2 tumors/colon in mice given fasudil vs 9 tumors/colon in mice given control agent). We made similar observations in IL10-deficient mice. CONCLUSIONS: We found inflammatory cytokines to reduce the activities of miRNAs. In mice with colitis, activities of miRNAs are reduced; administration of an agent that increases miRNA function prevents colon tumor formation in these mice. This pathway might be targeted to prevent colon carcinogenesis in patients with inflammatory bowel diseases.
  • Shiki Takamura; Hideki Yagi; Yoshiyuki Hakata; Chihiro Motozono; Sean R. McMaster; Tomoko Masumoto; Makoto Fujisawa; Tomomi Chikaishi; Junko Komeda; Jun Itoh; Miki Umemura; Ami Kyusai; Michio Tomura; Toshinori Nakayama; David L. Woodland; Jacob E. Kohlmeier; Masaaki Miyazawa
    JOURNAL OF EXPERIMENTAL MEDICINE 213 13 3057 - 3073 2016年12月 [査読有り]
     
    CD8(+) tissue-resident memory T cells (T-RM cells) reside permanently in nonlymphoid tissues and provide a first line of protection against invading pathogens. However, the precise localization of CD8(+) T-RM cells in the lung, which physiologically consists of a markedly scant interstitium compared with other mucosa, remains unclear. In this study, we show that lung CD8(+) T-RM cells localize predominantly in specific niches created at the site of regeneration after tissue injury, whereas peripheral tissue-circulating CD8(+) effector memory T cells (T-EM cells) are widely but sparsely distributed in unaffected areas. Although CD69 inhibited sphingosine 1-phosphate receptor 1-mediated egress of CD8(+) T cells immediately after their recruitment into lung tissues, such inhibition was not required for the retention of cells in the T-RM niches. Furthermore, despite rigid segregation of T-EM cells from the T-RM niche, prime-pull strategy with cognate antigen enabled the conversion from T-EM cells to T-RM cells by creating de novo T-RM niches. Such damage site-specific localization of CD8(+) T-RM cells may be important for efficient protection against secondary infections by respiratory pathogens.
  • 宮澤 正顯; 博多 義之; 武田 英里; 李 君; 河原 佐智代
    生化学 88 5 582 - 592 2016年 [査読有り]
  • Intracellular delivery of a peptide cargo by a cell-penetrating peptide via leucine-zippers does not affect the function of cargo
    Hakata; Y. Tsuchiya, S; Michiue, H; Ohtsuki, T; Matsui, H; Miyazawa M; Kitamatsu, M
    Chemical Communications 51 413 - 416 2015年05月
  • Y. Hakata; S. Tsuchiya; H. Michiue; T. Ohtsuki; H. Matsui; M. Miyazawa; M. Kitamatsu
    CHEMICAL COMMUNICATIONS 51 2 413 - 416 2015年 [査読有り]
     
    A hybrid comprising an autophagy-inducing peptide (AIP) and a cell-penetrating peptide (CPP) connected via heterodimeric leucine zippers was generated and delivered into cells. The hybrid successfully induced autophagy without significant cell death, while the same AIP directly connected to a CPP caused both autophagy and significant cell death.
  • Maiko Kato; Sachiyo Tsuji-Kawahara; Yuri Kawasaki; Saori Kinoshita; Tomomi Chikaishi; Shiki Takamura; Makoto Fujisawa; Akira Kawada; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 89 2 1468 - 1473 2015年01月 [査読有り]
     
    Toll-like receptor 7 and Myd88 are required for antiretroviral antibody and germinal center responses, but whether somatic hypermutation and class-switch recombination are required for antiretroviral immunity has not been examined. Mice deficient in activation-induced cytidine deaminase (AID) resisted Friend virus infection, produced virus-neutralizing antibodies, and controlled viremia. Passive transfer demonstrated that immune IgM from AID-deficient mice contributes to Friend virus control in the presence of virus-specific CD4(+) T cells.
  • Yoshiyuki Hakata; Masaaki Miyazawa; Nathaniel R. Landau
    VIROLOGY JOURNAL 11 108  2014年06月 [査読有り]
     
    Background: HIV-1 Vpr-mediated G(2) cell cycle arrest is dependent on the interaction of Vpr with an E3 ubiquitin ligase that contains damage-specific DNA binding protein 1 (DDB1), Cullin 4A (Cul4A), DDB1 and Cul4-associated factor 1 (DCAF1), and Rbx1. Vpr is thought to associate directly with DCAF1 in the E3 ubiquitin ligase complex although the exact interaction pattern of the proteins in the complex is not completely defined. The Vpr of SIVagm induces G(2) arrest of cognate African Green Monkey (AGM) cells but not human cells. The molecular mechanism by which SIVagm Vpr exhibits its species-specific function remained unknown. Methods: Physical interaction of proteins in the E3 ubiquitin ligase complex was assessed by co-immunoprecipitation followed by western blotting. In addition, co-localization of the proteins in cells was investigated by confocal microscopy. The cell cycle was analyzed by propidium iodide staining and flow cytometry. DNA damage response elicited by Vpr was evaluated by detecting phosphorylation of H2AX, a marker for DNA damage response. Results: We show that RNAi knock-down of DCAF1 prevented the co-immunoprecipitation of DDB1 with HIV-1 Vpr while DDB1 knock-down did not influence the binding of Vpr to DCAF1. HIV-1 Vpr mutants with a L64P or a R90K mutation maintained the ability to associate with DCAF1 but did not appear to be in a complex with DDB1. SIVagm Vpr associated with AGM DCAF1 and DDB1 while, in human cells, it binds to human DCAF1 but hardly binds to human DDB1, resulting in the reduced activation of H2AX. Conclusions: The identification of Vpr mutants which associate with DCAF1 but only poorly with DDB1 suggests that DCAF1 is necessary but the simple binding of Vpr to DCAF1 is not sufficient for the Vpr association with DDB1-containing E3 ligase complex. Vpr may interact both with DCAF1 and DDB1 in the E3 ligase complex. Alternatively, the interaction of Vpr and DCAF1 may induce a conformational change in DCAF1 or Vpr that promotes the interaction with DDB1. The ability of SIVagm Vpr to associate with DDB1, but not DCAF1, can explain the species-specificity of SIVagm Vpr-mediated G(2) arrest.
  • Sachiyo Tsuji-Kawahara; Shiki Takamura; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 88 9 5202 - 5203 2014年05月 [査読有り]
  • Shiki Takamura; Eiji Kajiwara; Sachiyo Tsuji-Kawahara; Tomoko Masumoto; Makoto Fujisawa; Maiko Kato; Tomomi Chikaishi; Yuri Kawasaki; Saori Kinoshita; Manami Itoi; Nobuo Sakaguchi; Masaaki Miyazawa
    PLOS PATHOGENS 10 3 e1003937  2014年03月 [査読有り]
     
    In chronic viral infections, persistent antigen presentation causes progressive exhaustion of virus-specific CD8(+) T cells. It has become clear, however, that virus-specific naive CD8(+) T cells newly generated from the thymus can be primed with persisting antigens. In the setting of low antigen density and resolved inflammation, newly primed CD8(+) T cells are preferentially recruited into the functional memory pool. Thus, continual recruitment of naive CD8(+) T cells from the thymus is important for preserving the population of functional memory CD8(+) T cells in chronically infected animals. Friend virus (FV) is the pathogenic murine retrovirus that establishes chronic infection in adult mice, which is bolstered by the profound exhaustion of virus-specific CD8(+) T cells induced during the early phase of infection. Here we show an additional evasion strategy in which FV disseminates efficiently into the thymus, ultimately leading to clonal deletion of thymocytes that are reactive to FV antigens. Owing to the resultant lack of virus-specific recent thymic emigrants, along with the above exhaustion of antigen-experienced peripheral CD8(+) T cells, mice chronically infected with FV fail to establish a functional virus-specific CD8(+) T cell pool, and are highly susceptible to challenge with tumor cells expressing FV-encoded antigen. However, FV-specific naive CD8(+) T cells generated in uninfected mice can be primed and differentiate into functional memory CD8(+) T cells upon their transfer into chronically infected animals. These findings indicate that virus-induced central tolerance that develops during the chronic phase of infection accelerates the accumulation of dysfunctional memory CD8(+) T cells.
  • Sachiyo Tsuji-Kawahara; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 88 3 1854 - 1855 2014年02月 [査読有り]
  • Sachiyo Tsuji-Kawahara; Hiroyuki Kawabata; Hideaki Matsukuma; Saori Kinoshita; Tomomi Chikaishi; Mayumi Sakamoto; Yuri Kawasaki; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 87 24 13760 - 13774 2013年12月 [査読有り]
     
    To assess the possible contribution of host immune responses to the exertion of Fv2-associated resistance to Friend virus (FV)-induced disease development, we inoculated C57BL/6 (B6) mice that lacked various subsets of lymphocytes with FV containing no lactate dehydrogenase-elevating virus. Fv2(r) B6 mice lacking CD4(+) T cells developed early polycythemia and fatal erythroleukemia, while B6 mice lacking CD8(+) T cells remained resistant. Erythroid progenitor cells infected with spleen focus-forming virus (SFFV) were eliminated, and no polycythemia was observed in B cell-deficient B6 mice, but they later developed myeloid leukemia associated with oligoclonal integration of ecotropic Friend murine leukemia virus. Additional depletion of natural killer and/or CD8(+) T cells from B cell-deficient B6 mice resulted in the expansion of SFFV proviruses and the development of polycythemia, indicating that SFFV-infected erythroid cells are not only restricted in their growth but are actively eliminated in Fv2r mice through cellular immune responses.
  • Tsumiyama K; Hashiramoto A; Takimoto M; Tsuji-Kawahara S; Miyazawa M; Shiozawa S
    Journal of immunology (Baltimore, Md. : 1950) 191 1 91 - 96 1 2013年07月 [査読有り]
     
    We investigated the role of effector CD8 T cells in the pathogenesis of immune glomerular injury. BALB/c mice are not prone to autoimmune disease, but after 12 immunizations with OVA they developed a variety of autoantibodies and glomerulonephritis accompanied by immune complex (IC) deposition. In these mice, IFN-γ-producing effector CD8 T cells were significantly increased concomitantly with glomerulonephritis. In contrast, after 12 immunizations with keyhole limpet hemocyanin, although autoantibodies appeared, IFN-γ-producing effector CD8 T cells did not develop, and glomerular injury was not induced. In β2-microglobulin-deficient mice lacking CD8 T cells, glomerular injury was not induced after 12 immunizations with OVA, despite massive deposition of IC in the glomeruli. In mice containing a targeted disruption of the exon encoding the membrane-spanning region of the Ig μ-chain (μMT mice), 12 immunizations with OVA induced IFN-γ- producing effector CD8 T cells but not IC deposition or glomerular injury. When CD8 T cells from mice immunized 12 times with OVA were transferred into naive recipients, glomerular injury could be induced, but only when a single injection of OVA was also given simultaneously. Importantly, injection of OVA could be replaced by one injection of the sera from mice that had been fully immunized with OVA. This indicates that deposition of IC is required for effector CD8 T cells to cause immune tissue injury. Thus, in a mouse model of systemic lupus erythematosus, glomerular injury is caused by effector CD8 T cells that recognize Ag presented as IC on the target renal tissue. Copyright © 2013 by The American Association of Immunologists, Inc.
  • 宮澤正顯
    ウイルス 62 1 27 - 38 1 2012年06月 [査読有り]
     
    APOBEC3は一本鎖DNAを標的とするシチジンデアミナーゼであり，レトロウイルス複製制限因子として機能する．現存の感染性レトロウイルスは自然宿主のAPOBEC3に対抗する機能を獲得しており，一方で宿主側も遺伝子重複によりAPOBEC3機能を進化させてきたと信じられていた．ところが最近，ヒトでもマウスでもAPOBEC3分子がタンパク質レベルで低発現となるような遺伝子多型が種の分岐後に獲得され，これが地理的に広範囲に分布している事実が明らかとなった．外来性の感染性レトロウイルスに対してAPOBEC3が示す生理的な防御効果と，感染性レトロウイルスの脅威が少ない場合に同じシチジンデアミナーゼが示す可能性のあるゲノムDNAの傷害作用との間に，微妙なバランスが存在するらしい．本稿ではAPOBEC3の生理機能と分子進化について最近の知見を解説した．
  • Naoto Hayasaka; Nobuo Nagai; Naoyuki Kawao; Atsuko Niwa; Yoshichika Yoshioka; Yuki Mori; Hiroshi Shigeta; Nobuo Kashiwagi; Masaaki Miyazawa; Takao Satou; Hideaki Higashino; Osamu Matsuo; Takamichi Murakami
    PLOS ONE 7 2 e32342  2012年02月 [査読有り]
     
    Background: There is an increasing need for animal disease models for pathophysiological research and efficient drug screening. However, one of the technical barriers to the effective use of the models is the difficulty of non-invasive and sequential monitoring of the same animals. Micro-CT is a powerful tool for serial diagnostic imaging of animal models. However, soft tissue contrast resolution, particularly in the brain, is insufficient for detailed analysis, unlike the current applications of CT in the clinical arena. We address the soft tissue contrast resolution issue in this report. Methodology: We performed contrast-enhanced CT (CECT) on mouse models of experimental cerebral infarction and hepatic ischemia. Pathological changes in each lesion were quantified for two weeks by measuring the lesion volume or the ratio of high attenuation area (%HAA), indicative of increased vascular permeability. We also compared brain images of stroke rats and ischemic mice acquired with micro-CT to those acquired with 11.7-T micro-MRI. Histopathological analysis was performed to confirm the diagnosis by CECT. Principal Findings: In the models of cerebral infarction, vascular permeability was increased from three days through one week after surgical initiation, which was also confirmed by Evans blue dye leakage. Measurement of volume and %HAA of the liver lesions demonstrated differences in the recovery process between mice with distinct genetic backgrounds. Comparison of CT and MR images acquired from the same stroke rats or ischemic mice indicated that accuracy of volumetric measurement, as well as spatial and contrast resolutions of CT images, was comparable to that obtained with MRI. The imaging results were also consistent with the histological data. Conclusions: This study demonstrates that the CECT scanning method is useful in rodents for both quantitative and qualitative evaluations of pathologic lesions in tissues/organs including the brain, and is also suitable for longitudinal observation of the same animals.
  • Jun Li; Yoshiyuki Hakata; Eri Takeda; Qingping Liu; Yasumasa Iwatani; Christine A. Kozak; Masaaki Miyazawa
    PLOS PATHOGENS 8 1 e1002478  2012年01月 [査読有り]
     
    Mouse apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like editing complex 3 (mA3), an intracellular antiviral factor, has 2 allelic variations that are linked with different susceptibilities to beta-and gammaretrovirus infections among various mouse strains. In virus-resistant C57BL/6 (B6) mice, mA3 transcripts are more abundant than those in susceptible BALB/c mice both in the spleen and bone marrow. These strains of mice also express mA3 transcripts with different splicing patterns: B6 mice preferentially express exon 5-deficient (Delta 5) mA3 mRNA, while BALB/c mice produce exon 5-containing full-length mA3 mRNA as the major transcript. Although the protein product of the Delta 5 mRNA exerts stronger antiretroviral activities than the full-length protein, how exon 5 affects mA3 antiviral activity, as well as the genetic mechanisms regulating exon 5 inclusion into the mA3 transcripts, remains largely uncharacterized. Here we show that mA3 exon 5 is indeed a functional element that influences protein synthesis at a post-transcriptional level. We further employed in vitro splicing assays using genomic DNA clones to identify two critical polymorphisms affecting the inclusion of exon 5 into mA3 transcripts: the number of TCCT repeats upstream of exon 5 and the single nucleotide polymorphism within exon 5 located 12 bases upstream of the exon 5/intron 5 boundary. Distribution of the above polymorphisms among different Mus species indicates that the inclusion of exon 5 into mA3 mRNA is a relatively recent event in the evolution of mice. The widespread geographic distribution of this exon 5-including genetic variant suggests that in some Mus populations the cost of maintaining an effective but mutagenic enzyme may outweigh its antiviral function.
  • Manuela Sironi; Franca Rosa Guerini; Cristina Agliardi; Mara Biasin; Rachele Cagliani; Matteo Fumagalli; Domenico Caputo; Andrea Cassinotti; Sandro Ardizzone; Milena Zanzottera; Elisabetta Bolognesi; Stefania Riva; Yasuyoshi Kanari; Masaaki Miyazawa; Mario Clerici
    MOLECULAR BIOLOGY AND EVOLUTION 28 12 3319 - 3329 2011年12月 [査読有り]
     
    The human RAC2 gene encodes a small GTP-binding protein with a pivotal role in immune activation and in the induction of peripheral immune tolerance through restimulation-induced cell death (RICD). Different human pathogens target the protein product of RAC2, suggesting that the gene may be subject to natural selection, and that variants in RAC2 may affect immunological phenotypes in humans. We scanned the genomic region encompassing the entire transcription unit for the presence of putative noncoding regulatory elements conserved across mammals. This information was used to select two RAC2 gene regions and analyze their intraspecific genetic diversity. Results suggest that a region covering the 3' untranslated region has been a target of multiallelic balancing selection (or diversifying selection), and three major RAC2 haplogroups occur in human populations. Haplotypes belonging to one of these clades are associated with increased susceptibility to multiple sclerosis (P = 0.022) and earlier onset of disease symptoms (P = 0.025). This same haplogroup is significantly more common in patients with Crohn's disease compared with healthy controls (P = 0.048). These data reinforce recent evidences that susceptibility alleles/haplotypes are shared among multiple autoimmune disorders and support a causal "role for RAC2" variants in the pathogenesis of autoimmune diseases. Other genes with a role in RICD have previously been associated with autoimmunity in humans, suggesting that this pathway and RAC2 may represent novel therapeutic targets in autoimmune disorders.
  • Tatsuya Ogawa; Sachiyo Tsuji-Kawahara; Takae Yuasa; Saori Kinoshita; Tomomi Chikaishi; Shiki Takamura; Haruo Matsumura; Tsukasa Seya; Toshihiko Saga; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 85 11 5423 - 5435 2011年06月 [査読有り]
     
    Natural killer (NK) cells function as early effector cells in the innate immune defense against viral infections and also participate in the regulation of normal and malignant hematopoiesis. NK cell activities have been associated with early clearance of viremia in experimental simian immunodeficiency virus and clinical human immunodeficiency virus type 1 (HIV-1) infections. We have previously shown that NK cells function as major cytotoxic effector cells in vaccine-induced immune protection against Friend virus (FV)-induced leukemia, and NK cell depletion totally abrogates the above protective immunity. However, how NK cells recognize retrovirus-infected cells remains largely unclear. The present study demonstrates a correlation between the expression of the products of retinoic acid early transcript-1 (RAE-1) genes in target cells and their susceptibility to killing by NK cells isolated from FV-infected animals. This killing was abrogated by antibodies blocking the NKG2D receptor in vitro. Further, the expression of RAE-1 proteins on erythroblast surfaces increased early after FV inoculation, and administration of an RAE-1-blocking antibody resulted in increased spleen infectious centers and exaggerated pathology, indicating that FV-infected erythroid cells are recognized by NK cells mainly through the NKG2D-RAE-1 interactions in vivo. Enhanced retroviral replication due to host gene-targeting resulted in markedly increased RAE-1 expression in the absence of massive erythroid cell proliferation, indicating a direct role of retroviral replication in RAE-1 upregulation.
  • Naoto Hayasaka; Kazuyuki Aoki; Saori Kinoshita; Shoutaroh Yamaguchi; John K. Wakefield; Sachiyo Tsuji-Kawahara; Kazumasa Horikawa; Hiroshi Ikegami; Shigeharu Wakana; Takamichi Murakami; Ram Ramabhadran; Masaaki Miyazawa; Shigenobu Shibata
    PLOS ONE 6 3 e17655  2011年03月 [査読有り]
     
    Regulators of G protein signaling (RGS) are a multi-functional protein family, which functions in part as GTPase-activating proteins (GAPs) of G protein alpha-subunits to terminate G protein signaling. Previous studies have demonstrated that the Rgs16 transcripts exhibit robust circadian rhythms both in the suprachiasmatic nucleus (SCN), the master circadian light-entrainable oscillator (LEO) of the hypothalamus, and in the liver. To investigate the role of RGS16 in the circadian clock in vivo, we generated two independent transgenic mouse lines using lentiviral vectors expressing short hairpin RNA (shRNA) targeting the Rgs16 mRNA. The knockdown mice demonstrated significantly shorter free-running period of locomotor activity rhythms and reduced total activity as compared to the wild-type siblings. In addition, when feeding was restricted during the daytime, food-entrainable oscillator (FEO)-driven elevated food-anticipatory activity (FAA) observed prior to the scheduled feeding time was significantly attenuated in the knockdown mice. Whereas the restricted feeding phase-advanced the rhythmic expression of the Per2 clock gene in liver and thalamus in the wild-type animals, the above phase shift was not observed in the knockdown mice. This is the first in vivo demonstration that a common regulator of G protein signaling is involved in the two separate, but interactive circadian timing systems, LEO and FEO. The present study also suggests that liver and/or thalamus regulate the food-entrained circadian behavior through G protein-mediated signal transduction pathway(s).
  • Taeko K. Naruse; Zhiyong Chen; Risa Yanagida; Tomoko Yamashita; Yusuke Saito; Kazuyasu Mori; Hirofumi Akari; Yasuhiro Yasutomi; Masaaki Miyazawa; Tetsuro Matano; Akinori Kimura
    IMMUNOGENETICS 62 9 601 - 611 2010年09月 [査読有り]
     
    Rhesus macaques (Macaca mulatta) are widely used in developing a strategy for vaccination against human immunodeficiency virus by using simian immunodeficiency virus infection as a model system. Because the genome diversity of major histocompatibility complex (MHC) is well known to control the immune responsiveness to foreign antigens, MHC loci in Indian- and Chinese-origin macaques used in the experiments have been characterized, and it was revealed that the diversity of MHC in macaques was larger than the human MHC. To further characterize the diversity of Mamu-A and Mamu-B loci, we investigated a total of 73 different sequences of Mamu-A, 83 sequences of Mamu-B, and 15 sequences of Mamu-I cDNAs isolated from Burmese-origin macaques. It was found that there were one to five expressing genes in each locus. Among the Mamu-A, Mamu-B, and Mamu-I sequences, 44 (60.2%), 45 (54.2%), and 8 (53.3%), respectively, were novel, and most of the other known alleles were identical to those reported from Chinese- or Indian-origin macaques, demonstrating a genetic mixture between the geographically distinct populations of present day China and India. In addition, it was found that a Mamu haplotype contained at least two highly transcribed Mamu-A genes, because multiple Mamu-A1 cDNAs were obtained from one haplotype. These findings further revealed the diversity and complexity of MHC locus in the rhesus macaques.
  • 成瀬 妙子; 陳 智勇; 柳田 梨紗; 山下 智子; 齋藤 祐介; 森 一泰; 保富 康宏; 宮澤 正顕; 俣野 哲朗; 木村 彰方
    MHC: Major Histocompatibility Complex 17 2 156 - 156 (一社)日本組織適合性学会 2010年08月
  • 奥田 裕紀子; 成瀬 妙子; 俣野 哲朗; 森 一泰; 保富 康宏; 宮澤 正顕; 木村 彰方
    MHC: Major Histocompatibility Complex 17 2 179 - 179 (一社)日本組織適合性学会 2010年08月
  • Takamura, S; M. Miyazawa
    J. Immunol. 185 3 1349 - 1350 2010年08月 [査読有り]
  • Chie Sugimoto; Satoru Watanabe; Taeko Naruse; Eiji Kajiwara; Teiichiro Shiino; Natsuko Umano; Kayoko Ueda; Hirotaka Sato; Shinji Ohgimoto; Vanessa Hirsch; Francois Villinger; Aftab A. Ansari; Akinori Kimura; Masaaki Miyazawa; Yasuo Suzuki; Naoki Yamamoto; Yoshiyuki Nagai; Kazuyasu Mori
    PLOS ONE 5 7 e11678  2010年07月 [査読有り]
     
    HIV vaccine development has been hampered by issues such as undefined correlates of protection and extensive diversity of HIV. We addressed these issues using a previously established SIV-macaque model in which SIV mutants with deletions of multiple gp120 N-glycans function as potent live attenuated vaccines to induce near-sterile immunity against the parental pathogenic SIVmac239. In this study, we investigated the protective efficacy of these mutants against a highly pathogenic heterologous SIVsmE543-3 delivered intravenously to rhesus macaques with diverse MHC genotypes. All 11 vaccinated macaques contained the acute-phase infection with blood viral loads below the level of detection between 4 and 10 weeks postchallenge (pc), following a transient but marginal peak of viral replication at 2 weeks in only half of the challenged animals. In the chronic phase, seven vaccinees contained viral replication for over 80 weeks pc, while four did not. Neutralizing antibodies against challenge virus were not detected. Although overall levels of SIV specific T cell responses did not correlate with containment of acute and chronic viral replication, a critical role of cellular responses in the containment of viral replication was suggested. Emergence of viruses with altered fitness due to recombination between the vaccine and challenge viruses and increased gp120 glycosylation was linked to the failure to control SIV. These results demonstrate the induction of effective protective immune responses in a significant number of animals against heterologous virus by infection with deglycosylated attenuated SIV mutants in macaques with highly diverse MHC background. These findings suggest that broad HIV cross clade protection is possible, even in hosts with diverse genetic backgrounds. In summary, results of this study indicate that deglycosylated live-attenuated vaccines may provide a platform for the elucidation of correlates of protection needed for a successful HIV vaccine against diverse isolates.
  • Sachiyo Tsuji-Kawahara; Tomomi Chikaishi; Eri Takeda; Maiko Kato; Saori Kinoshita; Eiji Kajiwara; Shiki Takamura; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 84 12 6082 - 6095 2010年06月 [査読有り]
     
    Several host genes control retroviral replication and pathogenesis through the regulation of immune responses to viral antigens. The Rfv3 gene influences the persistence of viremia and production of virus-neutralizing antibodies in mice infected with Friend mouse retrovirus complex (FV). This locus has been mapped within a narrow segment of mouse chromosome 15 harboring the APOBEC3 and BAFF-R loci, both of which show functional polymorphisms among different strains of mice. The exon 5-lacking product of the APOBEC3 allele expressed in FV-resistant C57BL/6 (B6) mice directly restricts viral replication, and mice lacking the B6-derived APOBEC3 exhibit exaggerated pathology and reduced production of neutralizing antibodies. However, the mechanisms by which the polymorphisms at the APOBEC3 locus affect the production of neutralizing antibodies remain unclear. Here we show that the APOBEC3 genotypes do not directly affect the B-cell repertoire, and mice lacking B6-derived APOBEC3 still produce FV-neutralizing antibodies in the presence of primed T helper cells. Instead, higher viral loads at a very early stage of FV infection caused by either a lack of the B6-derived APOBEC3 or a lack of the wild-type BAFF-R resulted in slower production of neutralizing antibodies. Indeed, B cells were hyperactivated soon after infection in the APOBEC3- or BAFFR-deficient mice. In contrast to mice deficient in the B6-derived APOBEC3, which cleared viremia by 4 weeks after FV infection, mice lacking the functional BAFF-R allele exhibited sustained viremia, indicating that the polymorphisms at the BAFF-R locus may better explain the Rfv3-defining phenotype of persistent viremia.
  • Shiki Takamura; Sachiyo Tsuji-Kawahara; Hideo Yagita; Hisaya Akiba; Mayumi Sakamoto; Tomomi Chikaishi; Maiko Kato; Masaaki Miyazawa
    JOURNAL OF IMMUNOLOGY 184 9 4696 - 4707 2010年05月 [査読有り]
     
    During chronic viral infection, persistent exposure to viral Ags leads to the overexpression of multiple inhibitory cell-surface receptors that cause CD8(+) T cell exhaustion. The severity of exhaustion correlates directly with the level of infection and the number and intensity of inhibitory receptors expressed, and it correlates inversely with the ability to respond to the blockade of inhibitory pathways. Friend virus (FV) is a murine retrovirus complex that induces acute high-level viremia, followed by persistent infection and leukemia development, when inoculated into immunocompetent adult mice. In this article, we provide conclusive evidence that FV infection results in the generation of virus-specific effector CD8(+) T cells that are terminally exhausted. Acute FV-induced disease is characterized by a rapid increase in the number of virus-infected erythroblasts, leading to massive splenomegaly. Most of the expanded erythroblasts strongly express programmed death ligand-1 and MHC class I, thereby creating a highly tolerogenic environment. Consequently, FV-specific effector CD8(+) T cells uniformly express multiple inhibitory receptors, such as programmed cell death 1 (PD-1), T cell Ig domain and mucin domain 3 (Tim-3), lymphocyte activation gene-3, and CTLA-4, rapidly become nonresponsive to restimulation and are no longer reinvigorated by combined in vivo blockade of PD-1 and Tim-3 during the memory phase. However, combined blockade of PD-1 and Tim-3 during the priming/differentiation phase rescued FV-specific CD8(+) T cells from becoming terminally exhausted, resulting in improved CD8(+) T cell functionality and virus control. These results highlight FV's unique ability to evade virus-specific CD8(+) T cell responses and the importance of an early prophylactic approach for preventing terminal exhaustion of CD8(+) T cells. The Journal of Immunology, 2010, 184: 4696-4707.
  • Hiroki Takakuwa; Toshiyuki Maruoka; Tadayo Hata; Masaaki Miyazawa; Tomoyo Hata; Hitoshi Toshimori; Koichi Otsuki
    Environmental Health and Preventive Medicine 15 2 121 - 123 2010年03月 [査読有り]
     
    Objectives: We evaluated the effectiveness and safety of a disinfectant newly developed by our laboratories for use against influenza viruses. Methods: The effectiveness of our new disinfectant against avian, swine and human influenza viruses was tested in ovo. The acute toxicity of this disinfectant to two different cultured cell lines was investigated. Results: This new disinfectant showed very strong anti-influenza viral activity in the in ovo tests. All of the influenza viruses tested were inactivated very quickly. Following exposure to the disinfectant, the infectivity of all viral strains tested had been eliminated within ≤10 min. The infectant showed a weak acute toxicity in vitro. Conclusion: This new disinfectant is expected to be useful for preventing viral infection during a new influenza pandemic. © 2009 The Japanese Society for Hygiene.
  • Masaaki Miyazawa; Sachiyo Tsuji-Kawahara; Tomomi Chikaishi; Maiko Kato; Shiki Takamura
    RETROVIROLOGY 6 Suppl 2 O9  2009年
  • Miyazawa, Masaaki; Lopalco, Lucia; Mazzotta, Francesco; Caputo; Sergio Lo; Veas, Francisco; Clerici, Mario; for the ESN; Study Group
    AIDS 23 2 161 - 175 2009年01月 [査読有り]
  • Eri Takeda; Sachiyo Tsuji-Kawahara; Mayumi Sakamoto; Marc-Andre Langlois; Michael S. Neuberger; Cristina Rada; Masaaki Miyazawa
    JOURNAL OF VIROLOGY 82 22 10998 - 11008 2008年11月 [査読有り]
     
    Several members of the apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like complex 3 (APOBEC3) family in primates act as potent inhibitors of retroviral replication. However, lentiviruses have evolved mechanisms to specifically evade host APOBEC3. Likewise, murine leukemia viruses ( MuLV) exclude mouse APOBEC3 from the virions and cleave virion-incorporated APOBEC3. Although the betaretrovirus mouse mammary tumor virus has been shown to be susceptible to mouse APOBEC3, it is not known if APOBEC3 has a physiological role in restricting more widely distributed and long-coevolved mouse gammaretroviruses. The pathogenicity of Friend MuLV (F-MuLV) is influenced by several host genes: some directly restrict the cell entry or integration of the virus, while others influence the host immune responses. Among the latter, the Rfv3 gene has been mapped to chromosome 15 in the vicinity of the APOBEC3 locus. Here we have shown that polymorphisms at the mouse APOBEC3 locus indeed influence F-MuLV replication and pathogenesis: the APOBEC3 alleles of F-MuLV-resistant C57BL/6 and -susceptible BALB/c mice differ in their sequences and expression levels in the hematopoietic tissues and in their abilities to restrict F-MuLV replication both in vitro and in vivo. Furthermore, upon infection with the pathogenic Friend virus complex, (BALB/c x C57BL/6) F 1 mice displayed an exacerbated erythroid cell proliferation when the mice carried a targeted disruption of the C57BL/6-derived APOBEC3 allele. These results indicate, for the first time, that mouse APOBEC3 is a physiologically functioning restriction factor to mouse gammaretroviruses.
  • 成瀬 妙子; 柳田 梨紗; 俣野 哲郎; 森 一泰; 保富 康宏; 宮澤 正顕; 木村 彰方
    MHC: Major Histocompatibility Complex 15 2 179 - 179 (一社)日本組織適合性学会 2008年08月
  • Masaaki Miyazawa; Sachiyo Tsuji-Kawahara; Yasuyoshi Kanari
    VACCINE 26 24 2981 - 2996 2008年06月 [査読有り]
     
    Several host genes control retroviral replication and pathogenesis. These include genes that directly affect the replication of retroviruses in target cells and those that control the host immune responses to the viral. antigens. Host genetic factors that affect retroviratal reptication and immune responses to the viratal antigens have been best studied in mouse models of Friend leukemia virus (FV) infection. Several genes Located within the major histocompatibitity comptex (MHC), along with a separate gene not Linked to the MHC, influence the host immune responses to FV antigens. The tatter, the Rfv3, regutates the production of virus-neutratizing antibodies, and thus affects the duration of viremia. T-cell responses to the viral epitopes are controlled by MHC class I and class 11 genotypes, and both CD8(+) and CD4(+) T-cells are required for spontaneous immune resistance to FV infection. When CD4(+) T-helper cells are efficiently primed with a viral epitope, however, CD8(+) T-cells are not required for immune protection against FV infection, white B cells are absolutely required. There are individuals who possess human immunodeficiency virus type I (HIV-1)-reactive IgA antibodies in their mucosal. secretions and show strong T-cell responses to HIV-1 antigens, even though they are negative for HIV-1 genome and HIV-1-reactive serum IgG. These HIV-1-exposed but uninfected individuals rarely possess resistance-associated alleles at known AIDS-restricting loci such as CCR5 Delta 32. Recent genetic analyses have indicated that a Large proportion of such exposed but uninfected individuals may share a common genetic background.(c) 2008 Elsevier Ltd. All rights reserved.
  • レトロウイルス感染と宿主因子：エイズ制圧を目指して
    宮澤正顕; 河原佐智代; 金成安慶; 武田英里; 坂本真由美; 阿部弘之; 木下さおり; 湯浅貴恵; 梶原栄二; 馬野奈津子
    近畿大医誌(Med. J. Kinki Univ.) 33 3 209 - 238 2008年
  • Taku Matsumoto; Tetsuo Ajiki; Eiji Kajiwara; Yoshiyasu Mita; Tsunenori Fujita; Haruki Morimoto; Masaaki Miyazawa; Yonson Ku
    JOURNAL OF GASTROENTEROLOGY 43 5 390 - 396 2008年 [査読有り]
     
    Background. Although bacterial translocation is a significant problem in patients with obstructive jaundice, how translocation is promoted in this situation is not clearly understood. We previously reported the recovery of gut mucosal T-lymphocyte numbers in jaundiced rats following internal biliary drainage. This suggests that bile in the intestinal lumen promotes T-lymphocyte redistribution into the gut mucosa. To test this hypothesis, we have examined the expression patterns of chemokines that play an important role in lymphocyte recruitment into the small intestine. Methods. Four groups of rats receiving one of the following surgical procedures were studied: a sham operation (SHAM), common bile duct ligation (CBDL), CBDL followed by external drainage, or CBDL followed by internal drainage. Expression levels of intestinal mRNAs encoding TECK, MECK, and LARC chemokines were assessed using real-time polymerase chain reaction. Distribution of chemokine mRNA in the rat ileum was examined using in situ hybridization (ISH). Results. Following surgery, the expression levels of TECK mRNA decreased significantly in the CBDL group compared with in the SHAM group. While TECK expression did not recover after external drainage, it recovered to a near-normal level after internal drainage. Expression levels of MECK and LARC mRNAs were similar among all groups. ISH confirmed strong expression of TECK mRNA in the epithelial cells of the small intestine. Conclusions. These results indicate that bile may contribute to high expression levels of TECK/CCL25 mRNA in the small intestine. Bile may also have a role in regulating the distribution of gut mucosal T lymphocytes by promoting TECK production from epithelial cells.
  • 志知 大輔; 成瀬 妙子; 日野原 邦彦; 森 一泰; 俣野 哲郎; 本多 三男; 保富 康宏; 宮澤 正顕; 木村 彰方
    MHC: Major Histocompatibility Complex 14 2 140 - 140 (一社)日本組織適合性学会 2007年08月
  • Mara Biasin; Luca Piacentini; Sergio Lo Caputo; Yasuyoshi Kanari; Giuliana Magri; Daria Trabattoni; Valentina Naddeo; Lucia Lopalco; Alberto Clivio; Eugenio Cesana; Francesca Fasano; Cristina Bergamaschi; Francesco Mazzotta; Masaaki Miyazawa; Mario Clerici
    JOURNAL OF INFECTIOUS DISEASES 195 7 960 - 964 2007年04月 [査読有り]
     
    Apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G (APOBEC3G), a human cytidine deaminase, is a potent inhibitor of HIV replication. To explore a possible role of this protein in modulating in vivo susceptibility to HIV infection, we analyzed APOBEC3G expression in HIV-exposed seronegative individuals, HIV-seropositive patients, and healthy control subjects. The results showed that the expression of APOBEC3G is significantly increased in peripheral blood mononuclear cells (PBMCs)-mainly CD14(+) cells-and in cervical tissues of HIV-exposed seronegative individuals. Higher APOBEC3G expression correlated with a reduced susceptibility of PBMCs to in vitro infection with the HIV-1(Ba-L) R5 strain. APOBEC3G could be important in modulating in vivo susceptibility to sexually transmitted HIV infection.
  • Yumiko Tanaka-Takahashi; Michio Yasunami; Taeko Naruse; Kunihiko Hinohara; Tetsuro Matano; Kazuyasu Mori; Masaaki Miyazawa; Mitsuo Honda; Yasuhiro Yasutomi; Yoshiyuki Nagai; Akinori Kimura
    ELECTROPHORESIS 28 6 918 - 924 2007年03月 [査読有り]
     
    The rhesus macaque exhibits individual differences in susceptibility and resistance to infectious agents such as simian immunodeficiency virus (SIV) under experimental conditions, and these may be genetically determined at least in part by major histocompatibility complex (MHC) class I polymorphism. Although the importance of defining MHC class I polymorphism is well recognized, development of a generic and comprehensive molecular typing method of MHC class I alleles of the rhesus macaque has been hampered because, during the evolution of this species, multiple copies of similar DNA sequences have been generated by duplication events including the coding sequences of Mamu-A and Mamu-B loci. We report here a newly developed reference strand-mediated conformation analysis (RSCA)-based typing method of multiple Mamu-A and Mamu-B cDNAs that allowed us to estimate the number of expressed alleles. This technique detected 1-7 Mamu-A signals and 2-12 Mamu-B signals in a single sample, indicating that the number of functional alleles may vary. By comparing the data from the parents with those from the descendants in the breeding colony, several MHC class I haplotypes consisting of variable numbers of functional Mamu-A and Mamu-B alleles could be assigned.
  • Hiroyuki Yamamoto; Miki Kawada; Tetsuo Tsukamoto; Akiko Takeda; Hiroko Igarashi; Masaaki Miyazawa; Taeko Naruse; Michio Yasunami; Akinori Kimura; Tetsuro Matano
    JOURNAL OF GENERAL VIROLOGY 88 2 652 - 659 2007年02月 [査読有り]
     
    The X4-tropic simian/human immunodeficiency virus (SHIV) 89.6P (or 89.6PD) causes rapid CD4(+) T-cell depletion leading to an acute crash of the host immune system, whereas pathogenic R5-tropic simian immunodeficiency virus (SIV) infection, like HIV-1 infection in humans, results in chronic disease progression in macaques. Recent pre-clinical vaccine trials inducing cytotoxic T lymphocyte (CTL) responses have succeeded in controlling replication of the former but shown difficulty in control of the latter. Analysis of the immune responses involved in consistent control of SHIV would contribute to elucidation of the mechanism for consistent control of SIV replication. This study followed up rhesus macaques that showed vaccine-based control of primary SHIV89.6PD replication and found that all of these controllers maintained viraemia control for more than 2 years. SHIV89.6PD control was observed in vaccinees of diverse major histocompatibility complex (MHC) haplotypes and was maintained without rapid selection of CTL escape mutations, a sign of particular CTL pressure. Despite the vaccine regimen not targeting Env, all of the SHIV controllers showed efficient elicitation of de novo neutralizing antibodies by 6 weeks post-challenge. These results contrast with our previous observation of particular MHC-associated control of SIV replication without involvement of neutralizing antibodies and suggest that vaccine-based control of SHIV89.6PD replication can be stably maintained in the presence of multiple functional immune effectors.
  • Yohei Kida; Sachiyo Tsuji-Kawahara; Valentina Ostapenko; Saori Kinoshita; Eiji Kajiwara; Hiroyuki Kawabata; Takae Yuasa; Iwao Nishide; Susumu Yukawa; Masakazu Ichinose; Masaaki Miyazawa
    CANCER IMMUNOLOGY IMMUNOTHERAPY 55 12 1459 - 1469 2006年12月 [査読有り]
     
    Hyperthermia (HT), in combination with other conventional therapeutic modalities, has become a promising approach in cancer therapy. In addition to heat-induced apoptosis, an augmented immunological effect is considered to be a benefit of hyperthermic treatment over chemo- or radiotherapy. Here, we investigated the effect of regional HT targeting the liver on immune cells, especially T cells and antigen-presenting cells, which are important in recognizing and eliminating tumor cells and pathogens such as viruses. In healthy volunteers exposed to such regional HT, both CD4(+) and CD8(+) T cells that express an activation marker CD69 increased transiently at I h post-treatment, with a subsequent decrease to base levels at 6 It after the treatment. At 24 h post-treatment, the percentage of CD69-positive cells significantly increased again but only among CD8(+) T cells. IFN-gamma production from PHA-stimulated peripheral blood mononuclear cells was gradually and significantly increased in the 2 days following the heating procedure, peaking at 36 h post-treatment. Furthermore, we found marked increases in plasma levels of IL-1 beta and IL-6 starting at 24 h post-treatment. With regard to the number of each leukocyte subpopulation, a transient and dramatic decrease in the number of a subset of monocytes, CD14(+) CD16(-) cells, was observed at 1 h after the hyperthermic treatment, suggesting that the regional HT aimed at the liver may have influenced the extravasation of blood monocytes. No significant changes in T-cell activities or monocyte counts were observed in the volunteers exposed to heating of the lungs or the legs. These results suggest that heating of the liver may efficiently induce cellular immune responses to liver cancers.
  • Mizuho Kajikawa; Tomohisa Baba; Utano Tomaru; Yutaka Watanabe; Satoru Koganei; Sachiyo Tsuji-Kawahara; Naoki Matsumoto; Kazuo Yamamoto; Masaaki Miyazawa; Katsumi Maenaka; Akihiro Shizu; Masanori Kasahara
    JOURNAL OF IMMUNOLOGY 177 5 3108 - 3115 2006年09月 [査読有り]
     
    MILL (MHC class I-like located near the leukocyte receptor complex) is a family of MHC class I-like molecules encoded outside the MHC, which displays the highest sequence similarity to human MICA/B molecules among known class I molecules. In the present study, we show that the two members of the mouse MILL family, MILL1 and MILL2, are GPI-anchored glycoproteins associated with beta(2)-microglobulin (beta(2)m) and that cell surface expression of MILL1 or MILL2 does not require functional TAP molecules. MILL1 and MILL2 molecules expressed in bacteria could be refolded in the presence of beta(2)m, without adding any peptides. Hence, neither MILL1 nor MILL2 is likely to be involved in the presentation of peptides. Immunohistochemical analysis revealed that MILL1 is expressed in a subpopulation of thymic medullary epithelial cells and a restricted region of inner root sheaths in hair follicles. The present study provides additional evidence that MILL is a class I family distinct from MICA/B.
  • 成瀬 妙子; 安波 道郎; 俣野 哲郎; 森 一泰; 本多 三男; 保富 康宏; 宮澤 正顕; 木村 彰方
    MHC: Major Histocompatibility Complex 13 2 117 - 117 (一社)日本組織適合性学会 2006年08月
  • H Kawabata; A Niwa; S Tsuji-Kawahara; H Uenishi; N Iwanami; H Matsukuma; H Abe; N Tabata; H Matsumura; M Miyazawa
    INTERNATIONAL IMMUNOLOGY 18 1 183 - 198 2006年01月 [査読有り]
     
    CD8(+) CTLs and virus-neutralizing antibodies have been associated with spontaneous and vaccine-induced immune control of retroviral infections. We previously showed that a single immunization with an env gene-encoded CD4(+) T cell epitope protected mice against fatal Friend retrovirus infection. Here, we analyzed immune cell components required for the peptide-induced anti-retroviral protection. Mice lacking CD8(+) T cells were nevertheless protected against Friend virus infection, while mice lacking B cells were not. Virus-producing cells both in the spleen and bone marrow decreased rapidly in their number and became undetectable by 4 weeks after infection in the majority of the peptide-immunized animals even in the absence of CD8(+) T cells. In the vaccinated animals the production and class switching of virus-neutralizing and anti-leukemia cell antibodies were facilitated; however, virus-induced erythroid cell expansion was suppressed before neutralizing antibodies became detectable in the serum. Further, the numbers of virus-producing cells in the spleen and bone marrow in the early stage of the infection were smaller in the peptide-immunized than in unimmunized control mice in the absence of B cells. Thus, peptide immunization facilitates both early cellular and late humoral immune responses that lead to the effective control of the retrovirus-induced disease, but CD8(+) T cells are not crucial for the elimination of virus-infected cells in the peptide-primed animals.
  • K Mori; C Sugimoto; S Ohgimoto; EE Nakayama; T Shioda; S Kusagawa; Y Takebe; M Kano; T Matano; T Yuasa; D Kitaguchi; M Miyazawa; Y Takahashi; M Yasunami; A Kimura; N Yamamoto; Y Suzuki; Y Nagai
    JOURNAL OF VIROLOGY 79 16 10386 - 10396 2005年08月 [査読有り]
     
    The envelope glycoprotein (Env) of human immunodeficiency viruses (HIVs) and simian immunodeficiency viruses (SIVs) is heavily glycosylated, and this feature has been speculated to be a reason for the insufficient immune control of these viruses by their hosts. In a macaque AIDS model, we demonstrated that quintuple deglycosylation in Env altered a pathogenic virus, SIVmac239, into a novel attenuated mutant virus (Delta 5G). In Delta 5G-infected animals, strong protective immunity against SIVmac239 was elicited. These HIV and SIV studies suggested that an understanding of the role of glycosylation is critical in defining not only the virological properties but also the immunogenicity of Env, suggesting that glycosylation in Env could be modified for the development of effective vaccines. To examine the effect of deglycosylation, we constructed prime-boost vaccines consisting of Env from SIVmac239 and Delta 5G and compared their immunogenicities and vaccine efficacies by challenge infection with SIVmac239. Vaccination-induced immune responses differed between the two vaccine groups. Both Env-specific cellular and humoral responses were higher in wild-type (wt)-Env-immunized animals than in Delta 5G Env-immunized animals. Following the challenge, viral loads in SIVmac239 Env (wt-Env)-immunized animals were significantly lower than in vector controls, with controlled viral replication in the chronic phase. Unexpectedly, viral loads in Delta 5G Env-immunized animals were indistinguishable from those in vector controls. This study demonstrated that the prime-boost Env vaccine was effective against homologous SIVmac239 challenge. Changes in glycosylation affected both cell-mediated and humoral immune responses and vaccine efficacy.
  • Y Kanari; M Clerici; H Abe; H Kawabata; D Trabattoni; S Lo Caputo; F Mazzotta; H Fujisawa; A Niwa; C Ishihara; YA Takei; M Miyazawa
    AIDS 19 10 1015 - 1024 2005年07月 [査読有り]
     
    Objective: Despite multiple and repeated exposures to HIV-1, some individuals possess no detectable HIV genome and show T-cell memory responses to the viral antigens. HIV-1-reactive mucosal IgA detected in such uninfected individuals suggests their possible immune resistance against HIV. We tested if the above HIV-1-exposed but uninfected status was associated with genetic markers other than a homozygous deletion of the CCR5 gene. Methods: Based on our mapping in chromosome 15 of a gene controlling the production of neutralizing antibodies in a mouse retrovirus infection, we genotyped 42 HIV-1-exposed but uninfected Italians at polymorphic loci in the syntenic segment of human chromosome 22, and compared them with 49 HIV-1-infected and 47 uninfected healthy control individuals by a closed testing procedure. Results: A significant association was found between chromosome 22q12-13 genotypes and a putative dominant locus conferring anti-HIV-1 immune responses in the exposed but uninfected individuals. Distributions of linkage disequilibrium across chromosome 22 also differed between the exposed but uninfected and two other phenotypic groups. Conclusions: The data indicated the presence of a new genetic factor associated with the HIV-1-exposed but uninfected status. (c) 2005 Lippincott Williams & Wilkins.
  • Miyazawa M
    Nihon rinsho. Japanese journal of clinical medicine 63 Suppl 5 217 - 222 2005年05月 [査読有り]
  • 松熊 秀明; 仲西 宏元; 河原 佐智代; 宮澤 正顯; 矢野 忠
    日本温泉気候物理医学会雑誌 68 3 181 - 188 The Japanese Society of Balneology, Climatology and Physical Medicine 2005年 
    Objectives—We tested the effect of a clinically applicable dose of moxibustion on adjuvant-induced arthritis (AIA) of rat, an experimental model of rheumatoid arthritis.
    Methods—Male Lewis rats were inoculated with Mycobacterium butyricum suspended in paraffin oil into the right hind paw to induce arthritis. Moxibustion (60°C) was applied to the right hind limb point, Tsu-san-Li (ST36), twice a week for 4 consecutive weeks. The efficacy of the above treatment was determined by the measurements of paw swelling, arthritic score. The effects of moxibustion upon immune and inflammatory responses were analyzed by enumerating peripheral blood leukocyte subsets. The data were analyzed with Mann-Whitney U-test between the experimental and control groups.
    Results—Moxibustion significantly suppressed paw swelling in the systemic phase, but not in the acute phase, of arthritis. Moxibustion also significantly suppressed the increase in T lymphocyte numbers in the late acute phase and that of neutroplils in the systemic phase.
    Conclusion—After the treatment with moxibustion, significant alterations were observed in the numbers of peripheral blood leukocyte subsets in AIA, along with the amelioration of clinical signs. These observations suggest that suppression of AIA with moxibustion may be mediated through the suppression of proliferating number of T-cell and acceleration of decrease in number of neutrophils in the peripheral blood.
  • D Sugahara; S Tsuji-Kawahara; M Miyazawa
    JOURNAL OF VIROLOGY 78 12 6322 - 6334 2004年06月 [査読有り]
     
    Recent studies have demonstrated an essential role of Gag-specific CD4(+) T-cell responses for viral control in individuals infected with human immunodeficiency virus type 1. However, little is known about epitope specificities and functional roles of the Gag-specific helper T-cell responses in terms of vaccine-induced protection against a pathogenic retroviral challenge. We have previously demonstrated that immunization with Friend murine leukemia virus (F-MuLV) Gag proteins protects mice against the fatal Friend retrovirus (FV) infection. We report here the structure of a protective T helper cell (Th) epitope, (I)VTWEAIAVDPPP, identified in the p15 (MA) region of F-MuLV Gag. In mice immunized with the Th epitope-harboring peptide or a vaccinia virus-expressed native full-length MA protein, FV-induced early splenomegaly regressed rapidly. In these mice, FV-infected cells were eliminated within 4 weeks and the production of virus-neutralizing antibodies was induced rapidly after FV challenge, resulting in strong protection against the virus infection. Interestingly, mice immunized with the whole NIA mounted strong CD4+ T-cell responses to the identified Th epitope, whereas mice immunized with mutant MA proteins that were not bound to the plasma membrane failed to mount efficient CD4+ T-cell responses, despite the presence of the Th epitope. These mutant MA proteins also failed to induce strong protection against FV challenge. These data indicate the importance of the properly processible NIA molecule for CD4+ T-cell priming and for the resultant induction of an effective immune response against retrovirus infections.
  • H Tahara; N Iwanami; N Tabata; H Matsumura; T Matsuura; T Kurita; M Miyazawa
    TRANSPLANT IMMUNOLOGY 13 1 25 - 32 2004年06月 [査読有り]
     
    In an MHC class I-disparate combination of mouse strains, a single intravenous injection of donor spleen cells combined with 10 suboptimal doses of 15-deoxyspergualin (DSG) administration was effective in inducing donor-specific suppression of cytotoxic T-lymphocyte (CTL) responses and prolonged survival of the relevant skin allograft. Proliferative potentials of the donor spleen cells were requirement for the induction of suppressed allospecific responses, but both highly purified T cells and non-T cells were equally effective to induce the suppression of CTL responses by intravenous injection. These results have shown that, although working on different mechanisms, DSG is as effective as FK506 or rapamycin in inducing allograft tolerance when used at suboptimal doses along with the donor-specific intravenous presensitization, and an immune mechanism other than well-characterized veto T cells is working in this model in suppressing alloreactive CTL precursors. (C) 2004 Elsevier B.V. All rights reserved.
  • Matano, T; M. Kobayashi; H. Igarashi; A. Takeda; H. Nakamura; M. Kano; C. Sugimoto; K. Mori; A. Iida; T. Hirata; M. Hasegawa; T. Yuasa; M. Miyazawa; Y. Takahashi; M. Yasunami; A. Kimura; D. H. O'Connor; D. I. Watkins; Y. Nagai
    The Journal of Experimental Medicine 199 12 1709 - 1718 2004年06月 [査読有り]
     
    Recently, encouraging AIDS vaccine trials in macaques have implicated cytotoxic T lymphocytes (CTLs) in the control of the simian human immunodeficiency virus SHIV89.6P that induces acute CD4(+) T cell depletion. However, none of these vaccine regimens have been successful in the containment of replication of the pathogenic simian immunodeficiency viruses (SIVs) that induce chronic disease progression. Indeed, it has remained unclear if vaccine-induced CTL can control SIV replication. Here, we show evidence suggesting that vaccine-induced CTLs control SIVmac239 replication in rhesus macaques. Eight macaques vaccinated with DNA-prime/Gag-expressing Sendai virus vector boost were challenged intravenously with SIVmac239. Five of the vaccinees controlled viral replication and had undetectable plasma viremia after 5 wk of infection. CTLs from all of these five macaques rapidly selected for escape mutations in Gag, indicating that vaccine-induced CTLs successfully contained replication of the challenge virus. Interestingly, analysis of the escape variant selected in three vaccinees that share a major histocompatibility complex class I haplotype revealed that the escape variant virus was at a replicative disadvantage compared with SIVmac239. These findings suggested that the vaccine-induced CTLs had "crippled" the challenge virus. Our results indicate that vaccine induction of highly effective CTLs can result in the containment of replication of a highly pathogenic immunodeficiency virus.
  • 宮澤正顯
    ウイルス 52 1 69 - 76 1 2002年06月 [査読有り]
  • J Sugita; H Ohtani; T Mizoi; K Saito; K Shiiba; Sasaki, I; S Matsuno; H Yagita; M Miyazawa; H Nagura
    JAPANESE JOURNAL OF CANCER RESEARCH 93 3 320 - 328 2002年03月 [査読有り]
     
    Anti-tumor immune responses are considered to be one of the key host reactions in human colorectal cancer, with T cells as important effector cells. For the induction of tumor-specific immunity, processing of cancer cells and priming of T cells by antigen-presenting cells are important. The present study was designed to clarify the relationship between Fas ligand (FasL; CD95L) expression and apoptotic cancer cells. Immunohistochemistry using frozen sections taken from 58 patients with colorectal cancer revealed that stromal cells composed mainly of tumor-associated macrophages expressed FasL, leaving cancer cells negative for FasL. These macrophages were abundantly distributed along the invasive margin. In situ hybridization revealed that these macro-phages as well as cancer cells expressed FasL mRNA, whereas macrophages in the normal colon mucosa rarely expressed FasL. Apoptotic cancer cells recognized by monoclonal antibody M30 CytoDEATH were localized not only in cancer cell nests, but also in the stroma along the invasive margin showing a dissociated pattern, which was particularly evident in the areas of FasL(+) macrophages. Furthermore, these macrophages, phenotypically similar to dendritic cells, occasionally contained M30(+) apoptotic cancer cells in the cytoplasm. Clinicopathologic analyses in 123 cases revealed 1) a positive correlation between the degree of dissociated M30(+) apoptotic cancer cells and the number of macrophages along the invasive margin and 2) an inverse association between the degree of dissociated M30(+) apoptotic cancer cells and the occurrence of hematogenous metastasis after surgical resection of the primary tumor. In conclusion, the present study shows the importance of FasL(+) activated macrophages as one of the host defense mechanisms against cancer cell spread in human colorectal cancer.
  • K Fukuoka; T Ajiki; M Miyazawa; Y Takeyama; H Onoyama; Y Kuroda
    EUROPEAN JOURNAL OF SURGERY 167 9 684 - 688 2001年09月 [査読有り]
     
    Objective: To examine the changes in the number of T cells and macrophages in the mucosal lamina propria in the presence or absence of bile in the gastrointestinal tract. Design: Clinical study. Setting: University hospital, Japan. Subjects: 6 patients with obstructive jaundice who had external biliary drainage (drainage group) and 6 patients with no signs of obstructive jaundice (control group). Interventions: Gastrointestinal specimens were taken at the time of operation. Main outcome Measures: The number of CD4(+) T cells, CD8(+) T cells and CD68(+) macrophages in the lamina propria mucosae in each group measured immunohistochemically. Results: The numbers of CD8+ T cells and CD68(+) macrophages in the lamina propria of the patients treated by external drainage were significantly less than in the control group (p < 0.01). However, there was no difference in the number of CD4+ T cells between the groups (p = 0.45). Conclusions: In the absence of bile, mucosal immune function fails as a result of reduced numbers of CD8+ T cells and macrophages.
  • Iwanami, N; A. Niwa; Y. Yasutomi; N. Tabata; M. Miyazawa
    Journal of Virology 75 7 3152 - 3163 2001年04月 [査読有り]
     
    We have previously shown that immunization with a synthetic peptide that contains a single CD4(+) T-cell epitope protects mice against immunosuppressive Friend retrovirus infection, Cells producing infectious Friend virus were rapidly eliminated from the spleens of mice that had been immunized with the single-epitope peptide. However, actual effector mechanisms induced through T-helper-cell responses after Friend virus inoculation were unknown. When cytotoxic effector cells detected in the early phase of Friend retrovirus infection were separated based on their expression of cell surface markers, those lacking CD4 and CD8 but expressing natural killer cell markers were found to constitute the majority of effector cells that lysed Friend virus-induced leukemia cells. Depletion of natural killer cells by injecting anti-asialo-ganglio-N-tetraosylceramide antibody did not affect the number of CD4(+) or CD8(+) T cells in the spleen, virus antigen-specific proliferative responses of CD4(+) T cells, or cytotoxic activity against Friend virus-induced leukemia cells exerted by CD8(+) effector cells. However, the same treatment markedly reduced the killing activity of CD4(-)CD8(-) effector cells and completely abolished the effect of peptide immunization, Although the above enhancement of natural killer cell activity in the early stage of Friend virus infection was also observed in mice given no peptide, these results have demonstrated the importance and requirement of natural killer cells in vaccine-induced resistance against the retroviral infection.
  • Kanamasa, Ken; Otani, Narutaka; Ishida, Norihiro; Inoue, Yoshikazu; Ikeda, Akiko; Morii, Hideki; Naito, Norikatsu; Hayashi, Takahiro; Ishikawa, Kinji; Miyazawa, Masaaki
    Journal of Cardiovascular Pharmacology 37 2 155 - 162 2001年02月 [査読有り]
     
    Thrombus formation is a key component of the pathogenesis of restenosis after arterial balloon injury. The purpose of this study was to determine whether intimal hyperplasia could be attenuated by infusion of recombinant tissue plasminogen activator (tPA). Forty-two Kurosawa and Kusanagi hypercholesterolemic rabbits were divided into tPA (n = 20) and control (n = 22) groups, the former receiving 7 days of continuous tPA infusion (0.6 mg/kg/day) via ear veins. The walls of the common iliac arteries were injured using 2.5-mm balloon catheters and then examined histologically 7, 14, 21, and 28 days later. Cell proliferation was assessed by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), and transforming growth factor (TCF)-beta immunohistochemistry was carried out to estimate cell proliferation and differentiation. It was observed that 28 days after balloon injury, intimal cross-sectional areas in the tPA group were significantly smaller than in controls (0.11 +/- 0.03 mm(2) vs. 0.57 +/- 0.08 mm(2), p < 0.01), as were ratios of the cross-sectional areas of the intima and media (0.21 +/- 0.07 vs. 1.06 +/- 0.18, p < 0.05). In addition, the numbers of PCNA-positive medial cells were significantly lower (0.06 +/- 0.01 vs. 0.36 +/- 0.08, p < 0.05) and TGF-<beta>-positive vessel wall areas were significantly smaller in tPA-treated animals 7 days after balloon injury (0.47 +/- 0.28% vs. 4.55 +/- 1.44%, p < 0.05). Thus infusion of tPA after arterial balloon injury appears to decrease medial cell proliferation and suppress intimal hyperplasia.
  • M Izuma; K Kobayashi; M Shiina; Y Ueno; M Ishii; T Shimosegawa; T Toyota; K Kakimi; M Miyazawa
    HEPATOLOGY RESEARCH 18 3 218 - 229 2000年11月 [査読有り]
     
    We investigated the responses of peripheral blood mononuclear cells (PBMCs) to hepatitis C virus core protein in ten patients with chronic hepatitis C during interferon (IFN)-beta treatment to determine if the modulation of the immune reaction to hepatitis C virus by IFN treatment is associated with the viral clearance. Interleukin-2, interleukin-4, interleukin-10, and interferon-gamma in the supernatant of the patients' PBMC co-cultured with the MCV core antigen-presenting autologous PBMCs were measured by ELISA. Serum levels of soluble CD (sCD) 8 and sCD30 in these patients were also measured by ELISA. The production of interleukin-2 and interferon-gamma by PBMCs of sustained responders (SRs) increased after IFN-treatment, although it did not reach a significant level. Interleukin-10 was detected only in non-responders (NRs) at 0 and 4 weeks after the start of IFN treatment. Serum sCD8 level increased significantly in SR with IFN treatment. A close correlation between the serum sCD8 levels and interferon-gamma levels in the supernatant at week 8 was observed in SR. These results suggest that IFN treatment potentiates the cellular immune reaction against HCV core protein more efficiently in SR than in NR. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.
  • M Miyazawa; N Tabata; R Fujisawa; K Hashimoto; H Shiwaku; YA Takei
    INTERNATIONAL JOURNAL OF CARDIOLOGY 75 Suppl 1 S65 - S73 2000年08月 [査読有り]
     
    MRL/MpJ-lpr/lpr (MRL/lpr) mice spontaneously develop immune complex-mediated glomerulonephritis, granulomatous arteritis, and thrombocytopenia. Recent genetic analyses in a few different strains of lupus-prone mice have pointed out a close correlation between autoantibodies reactive with the endogenous retroviral env gene product, gp70, and the development and severity of glomerulonephritis. We have also shown that autoantibodies reactive with endogenous retroviral gp70 are closely correlated with the development of necrotizing polyarteritis in another lupus-prone strain of mice, SL/Ni. However, suggested pathogenicity of anti-gp70 autoantibodies has not yet been directly tested. To examine if anti-gp70 autoantibodies induce glomerular and vascular pathology, we established from unmanipulated MRL/lpr mice hybridoma clones that secrete monoclonal antibodies reactive with endogenous xenotropic viral env gene products. As reported separately, a high proportion of these anti-gp70 antibody-producing hybridoma clones induced in syngeneic non-autoimmune and severe combined immunodeficiency mice proliferative or wire loop-like glomerular lesions with granular deposits of gp70, IgG, and C3 in affected glomeruli. Some mice transplanted with these anti-gp70 autoantibody-producing hybridoma cells also showed massive subendothelial deposition of electron-dense materials in small arterioles in the kidneys. Furthermore, we identified an IgG2a-producing anti-gp70 hybridoma clone that induced microvascular intraluminal platelet aggregation, thrombocytopenia, and amenia upon transplantation into syngeneic non-autoimmune mice. This anti-gp70 autoantibody bound onto the surfaces of mouse platelets, and specifically precipitated a platelet protein with an approximate relative molecular mass of 40 000. Attachment of activated platelets to the intimal surfaces of small arteries was also observed by electron microscopy in mice transplanted with the pathogenic anti-gp70 IgG2a-producing hybridoma cells, suggesting an interaction between antibody-bound platelets and endothelial cells. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.
  • Tabata, N; M. Miyazawa; R. Fujisawa; Y. A. Takei; H. Abe; K. Hashimoto
    Journal of Virology 74 9 4116 - 4126 2000年05月 [査読有り]
     
    Several strains of mice, including MRL/MpJ mice homozygous for the Fas mutant lpr gene (MRL/lpr mice), F(1) hybrids of New Zealand Black and New Zealand White mice, and BXSB/MpJ mice carrying a Y-linked autoimmune acceleration gene, spontaneously develop immune complex-mediated glomerulonephritis. The involvement of the envelope glycoprotein gp70 of an endogenous xenotropic virus in the formation of circulating immune complexes and their deposition in the glomerular lesions have been demonstrated, as has the pathogenicity of various antinuclear, antiphospholipid, and rheumatoid factor autoantibodies. In recent genetic linkage studies as well as in a study of cytokine-induced protection against nephritis development, the strongest association of serum levels of gp70-anti-gp70 immune complexes, rather than the levels of antinuclear autoantibodies, with the development and severity of glomerulonephritis has been demonstrated, suggesting a major pathogenic role of anti-gp70 autoantibodies in the lupus-prone mice, However, the pathogenicity of anti-gp70 autoantibodies has not yet been directly tested. To examine if anti-gp70 autoantibodies induce glomerular pathology, we established from unmanipulated MRL/lpr mice hybridoma clones that secrete monoclonal antibodies reactive with endogenous xenotropic viral Env gene products. Upon transplantation, a high proportion of these anti-gp70 antibody-producing hybridoma clones induced in syngeneic non-autoimmune and severe combined immunodeficiency mice proliferative or wire loop-like glomerular lesions. Furthermore, deposition of gp70 in glomeruli and pathological changes were observed after intravenous injection of representative clones of purified anti-gp70 immunoglobulin G, demonstrating pathogenicity of at least some anti-gp70 autoantibodies.
  • K Hashimoto; N Tabata; R Fujisawa; H Matsumura; M Miyazawa
    CLINICAL AND EXPERIMENTAL IMMUNOLOGY 119 1 47 - 56 2000年01月 [査読有り]
     
    MRL/MpJ-lpr/lpr (MRL/lpr) mice spontaneously develop immune complex-mediated glomerulonephritis and thrombocytopenia. Although the presence of cross-reactive anti-phospholipid antibodies in sera of MRL/lpr mice has been demonstrated, possible relationships between detected autoantibodies and the development of thrombocytopenia have not been elucidated. Recent genetic analyses in a few different strains of lupus-prone mice have pointed out a close correlation between autoantibodies reactive with endogenous retroviral env gene product, gp70, and the development and severity of glomerulonephritis. In the process of establishing possibly nephritogenic anti-gp70 autoantibody-producing hybridoma cells from MRL/lpr mice, we identified an IgG2a-producing anti-gp70 hybridoma clone that induced microvascular intraluminal platelet aggregation, thrombocytopenia, and amenia upon transplantation into syngeneic non-autoimmune mice. This and two other anti-gp70 antibodies bound onto the surface of mouse platelets, and purified IgG2a of the anti-gp70 autoantibody induced glomerular lesions with characteristics of thrombotic thrombocytopenic purpura when injected into non-autoimmune mice. The pathogenic anti-gp70 autoantibody specifically precipitated a platelet protein with an approximate relative molecular mass of 40 000.
  • M Miyazawa; Y Yanai; M Kurimoto
    JOURNAL OF HEPATOLOGY 31 5 967 - 967 1999年11月 [査読有り]
  • Hiroyuki Okuda; Masaaki Adachi; Masaaki Miyazawa; Yuji Hinoda; Kohzoh Imai
    Oncogene 18 40 5604 - 5609 1999年09月 [査読有り]
     
    Disruption of interactions between epithelial cells and extracellular matrix proteins leads to apoptosis of the cells, a phenomenon termed anoikis, Anoikis seems to play important roles in control of cellular positioning and inhibition of inappropriate cell growth, Here are found that a protein kinase C (PKC) activator phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA) promoted cell death in human gastric cancer cell lines MKN45 and MKN74 only when they lost anchorage. Loss of anchorage slightly increased enzymatic activity of PKC alpha, and an addition of TPA promoted cell death with further increase of PKC alpha activity, but not PKC beta in MKN45 cells, implicating an involvement of PKC alpha in anoikis, Furthermore, vaccinia virus-mediated overexpression of PKC alpha strongly increased CPP32 activity in the detached MKN45 and MKN74 cells, and augmented anoikis, however it had little effect on viability and CPP32 activity in the attached cells. Taken together, PKC alpha promotes apoptotic cell death in gastric cancer cells depending upon loss of anchorage, thereby may be a modulator of anoikis.
  • Sakamoto M; Miyazawa M; Mori S; Fujisawa R
    Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology 28 1 20 - 25 1999年01月 [査読有り]
     
    To test whether the autoantibodies reactive with epithelial cells of the salivary gland in sera from Sjogren's syndrome (SS) patients are specific for the organ and the disease, tissue reactivities of serum IgG obtained from the patients with SS and oral lichen planus (OLP), another immune-mediated oral mucosal disease, were examined by immunohistochemistry and Western blotting. IgG purified from the sera of SS patients specifically localized not only on the nuclei but also on the cytoplasm of the salivary gland epithelial cells. On the other hand, no convincing staining of the epithelial cells was observed when IgG purified from the sera of OLP patients or those from healthy controls were used for immunohistochemistry. No cytoplasmic staining was observed when sections of kidneys and pancreas were stained with SS patients' IgG. In Western blotting performed by using lysates of a submandibular gland as antigens, all the IgG prepared from the SS patients reacted prominently with several protein bands, including those specific for the disease and the organ. These results suggest that production of autoantibodies reacting with the cytoplasm of salivary gland epithelial cells is a characteristic of SS, and may play a role in the pathogenesis of the sialadenitis.
  • H Iijima; M Miyazawa; J Sakai; K Magoori; MR Ito; H Suzuki; M Nose; Y Kawarabayasi; TT Yamamoto
    JOURNAL OF BIOCHEMISTRY 124 4 747 - 755 1998年10月 [査読有り]
     
    The very low density lipoprotein receptor (VLDLR) gene contains an exon encoding a region of clustered serine and threonine residues immediately outside the membrane-spanning sequence, and this region has been proposed to be the site of clustered O-linked carbohydrate chains, Two forms of VLDLR transcripts, with and without the O-linked sugar region, are generated through alternative splicing. Reverse transcription polymerase chain reaction with RNAs from various rabbit tissues revealed that the VLDLR transcript with the O-linked sugar region (type-1 VLDLR) is the major transcript in heart and muscle, while the VLDLR transcript without the O-linked sugar region (type-2 VLDLR) predominates in non-muscle tissues, including cerebrum, cerebellum, kidney, spleen, adrenal gland, testis, ovary, and uterus, Hamster fibroblasts expressing type-a VLDLR bound with relatively low affinity to beta-migrating very low density lipoprotein compared with type-1 VLDLR-transfected cells, In contrast, the internalization, dissociation, and degradation of the ligand were not significantly impaired in either type of VLDLR-transfected cell. The receptor proteins in type-2 VLDLR-transfected cells underwent rapid degradation and accumulated in the culture medium, while those in type-1 VLDLR-transfected cells were stable and resistant to proteolytic cleavage. Analysis of the O-linked sugars of both types of transfected cells suggested that the O-linked sugar region is the major site for O-glycosylation.
  • H Tsumura; M Miyazawa; S Ogawa; JZ Wang; Y Ito; K Shimura
    LABORATORY ANIMALS 32 1 86 - 94 1998年01月 [査読有り]
     
    We characterized C-type retroviruses expressed in the pancreatic p-cells of non-obese diabetic (NOD) mice by immunohistochemical techniques and by inhibiting the production of viral particles using antisense oligonucleotides. Some cells in the pancreatic islets from both NOD and diabetes-resistant NOD-related mice (NON) reacted with a monoclonal antibody directed against the envelope protein(s) of polytropic viruses. On the other hand, NOD islet cells also showed strong immunoreactivity with an anti-gag protein monoclonal antibody and another anti-envelope protein(s) monoclonal antibody that is specific for xenotropic vises. In antisense oligodeoxynucleotide inhibition assays, a xenotropic virus-specific phosphorothionate antisense oligodeoxynucleotide significantly inhibited the occurrence of C-type virus particles in NOD mouse islet beta-cells. Therefore, C-type retrovirus-like particles expressed in NOD mouse pancreatic beta-cells were considered to be endogenous xenotropic virus. The expression of the xenotropic viral genome may be involved in the pathogenesis of the diabetic syndrome in NOD mice.
  • Miyazawa M
    Nihon rinsho. Japanese journal of clinical medicine 55 6 1370 - 1376 1997年06月 [査読有り]
  • Masaaki Miyazawa
    Leukemia 11 3 227 - 229 1997年 [査読有り]
     
    Friend murine retrmirus complex induces acute and fatal erythroleukemia when inoculated into immunocompelent adult mice. The development of leukemia after inoculation of Friend virus complex is controlled by several host genes. Some of the host genes influence immune responses against the viral antigens. Both CD4-positive T helper cells and CDS-positive cytotoxic T-lymphocytes specific for Friend viral antigens are required for spontaneous resistance against the virally induced leukemia. We have identified two separate T helper cell epitopes in the gp70 envelope glycoprotein encoded by the helper component of Friend virus complex. Immunization of mice with a synthetic peptide that represented one of the two T helper cell epitopes by a single injection with an adjuvant induced potent protective immunity against Friend virus-induced leukemia, even in the absence of CD8-positive T lymphocytes. In the immunized mice, virus-infected erythroid progenitor cells were rapidly eliminated from the spleen within two weeks after inoculation of the Friend virus. These data indicate unexpected importance and efficacy of CD4-positive T helper cells in immunity against relrovirus infections.
  • H Kumamoto; M Miyazawa; K Ooya
    JOURNAL OF ORAL PATHOLOGY & MEDICINE 25 9 484 - 490 1996年10月 [査読有り]
     
    To obtain monoclonal antibodies reactive with odontogenic but not other types of epithelium, mice were immunized with homogenates of fixed ameloblastoma tissues, and monoclonal antibodies Y4 and M11 were produced. Y4 reacted immunohistochemically with odontogenic epithelial components but not with those of squamous differentiation, while M11 reacted with odontogenic epithelial components and a part of keratotic epithelial tissues. Immunoglobulin isotypes of both antibodies were IgM as determined by Ouchterlony immunodiffusion and enzyme-linked immunosorbent assays, Western blotting revealed that the antigen recognized by Y4 had a molecular mass of approximately 66 kDa; however, the antigen reactive with M11 was not identified by Western blotting in spite of various attempts in changing reaction conditions. These antibodies may be beneficial to histological analyses of odontogenic tissues and their related lesions.
  • M Nose; MR Ito; M Ono; S Terasaki; M Miyazawa; S Mori
    INTERNATIONAL JOURNAL OF CARDIOLOGY 54 S35 - S44 1996年08月 [査読有り]
     
    The lpr and gld genes are thought to result in an incapacity for Fas-mediated apoptosis of T and B cells and the development of subsequent autoimmune disease. A newly established gld-congenic strain of mice, MRL/MpTn-gld/gld (MRL/gld), was found to develop vascular lesions involving arteritis and glomerulonephritis (GN), which were similar to those observed in the MRL/Mp-lpr/lpr (MRL/lpr) strain. However, comparative studies with a C3H/HeJ strain bearing lpr or gld revealed that these lesions developed only in mice with an MRL background. We were successful in transferring GN to normal MHC-compatible gld/gld and irradiated +/+ mice by bone marrow cells of MRL/gld mice, but were unsuccessful using those of C3H/gld mice. Transfer of arteritis, however, was only successful in mice with an MRL background. Nephritogenic monoclonal antibodies obtained from an MRL/lpr and an MRL/gld mouse were shown to be bone marrow-derived and rich in clonal diversity, and at least two of these were capable of causing glomerular injury by different mechanisms. Development of GN and systemic arteritis in MRL/lpr and MRL/gld mice will be dependent not only on their incapacity for Fas-mediated apoptosis but also on bone marrow cells and peripheral cells with intrinsic defects.
  • T Kazama; M Miyazawa; S Tsuchiya; A Horii
    BONE MARROW TRANSPLANTATION 18 2 437 - 441 1996年08月 [査読有り]
     
    A case of proliferation of CD68-positive macrophage-lineage cells in the bone marrow accompanied by severe thymic atrophy associated with graft-versus-host disease (GVHD) in a boy given allogeneic bone marrow transplantation (BMT) is reported, A 7-year-old boy was treated for posthepatitic severe aplastic anemia by BMT from his HLA-identical, mixed lymphocyte reaction-negative sister, After the transplantation his peripheral blood group converted to the donor type, However, the patient suffered from acute and chronic GVHD and slowly progressive anemia, and he died of multiple organ failure 21 months after BMT, At the autopsy diffuse and monotonous proliferation of CD68-positive macrophage-lineage cells was found to be replacing the blood-forming cells in the bone marrow, The thymus was almost empty of T lymphocytes, and remaining strands of extremely atrophic epithelial cells showed focal cystic change. Extramedullary hematopoiesis was found in the spleen, Analyses of microsatellite markers suggested the hematopoietic cells in the spleen to be of donor origin.
  • Y Saiki; H Ohtani; Y Naito; M Miyazawa; H Nagura
    LABORATORY INVESTIGATION 75 1 67 - 76 1996年07月 [査読有り]
     
    A subset of gastric carcinoma carries Epstein-Barr virus (EBV). The immunophenotypic features of EBV-associated (EBV(+)) gastric carcinoma, which we have analyzed using 25 EBV(+) cases, remain unclear. Frozen tissue samples were stained with antibodies to various immune cell markers. To evaluate the proliferative activity of CD8(+) cells, we performed CD8/Ki-67 double staining on paraffin-embedded sections. The results were compared with those in EBV-negative (EBV(-)) gastric carcinomas. All EBV(+) and EBV(-) gastric carcinoma cells expressed major histocompatibility complex class I, whereas major histocompatibility complex class II expression in tumor cells was more prominent in EBV(+) cases. Intercellular adhesion molecule-1 and Fas/APO-1 expression was largely restricted to EBV(+) cases. The lymphocytes that infiltrated EBV(+) tumor nests were predominantly CD8(+) T cells, many of which expressed perforin. Immunoelectron microscopy confirmed a close cell to cell contact between these CD8(+) cells and carcinoma calls. CD8(+) cells were CD11a(+) and CD11b(-) by flow cytometry performed in one case. The labeling index of Ki-67, the proliferation-associated antigen, in CD8(+) cells was 4 times higher in EBV(+) cases than in EBV(-) cases. Our data suggest that these CD8(+) cells, which bear a cytotoxic phenotype, are actively proliferating in close contact with EBV(+) tumor cells and that the specificity of the CD8(+) cells may be directed to EBV and/or cellular antigens expressed by the tumor. This is consistent with a generally favorable prognosis of EBV(+) gastric carcinoma. Because the observed T-cell infiltration is insufficient to eradicate the tumor cells, certain immunosuppressive factors were speculated to allow the essentially immunogenic carcinoma cells to establish a macroscopic lesion.
  • T KONDO; H UENISHI; T SHIMIZU; T HIRAMA; M IWASHIRO; K KURIBAYASHI; H TAMAMURA; N FUJII; R FUJISAWA; M MIYAZAWA; H YAMAGISHI
    JOURNAL OF VIROLOGY 69 11 6735 - 6741 1995年11月 [査読有り]
     
    Several dominant T-cell receptors of cytotoxic T-lymphocyte (CTL) clones specific for FBL-3 tumor antigen were clonally amplified in mixed lymphocyte tumor cell cultures derived from an individual immune mouse. Every CTL clone analyzed had a common specificity for a single epitope in the precursor to cell membrane associated nonstructural gag-encoded protein, pr75(gag), which can be minimally identified by nine amino acid residues, SIVLCCLCL. This epitope is located within the hydrophobic signal sequence motif that mediates translocation of the protein into the endoplasmic reticulum. These novel observations suggest that expression of Pr75(gag) in FBL-3 tumor cells led to the amplification of CTLs which recognize the signal sequence of the nonstructural gag-encoded glycoprotein precursor.
  • M MIYAZAWA; R FUJISAWA; C ISHIHARA; YA TAKEI; T SHIMIZU; H UENISHI; H YAMAGISHI; K KURIBAYASHI
    JOURNAL OF IMMUNOLOGY 155 2 748 - 758 1995年07月 [査読有り]
     
    Synthetic peptide vaccines containing a single Th cell epitope identified in the gp70 envelope glycoprotein of Friend murine leukemia helper virus induced potent protective immunity against Friend virus infection. H-2(a/b) mice immunized by a single s.c. injection of the CFA emulsion containing a peptide that represented the N-terminal gp70 epitope recovered slowly from initial development of splenomegaly, and most did not develop late leukemia, whereas most of the control mice given an injection of CFA alone showed sustained leukemic splenomegaly after the challenge with Friend virus. The mice of the same genetic background immunized with the C-terminal Th cell epitope by a single injection of a separate synthetic peptide eliminated virus-producing cells from the spleen within 12 days after inoculation of Friend virus complex, and did not develop early splenomegaly or polycythemia. H-2(a/a) mice were not protected by immunization with either one of the two synthetic peptides. Earlier production and more rapid class switching of virus-neutralizing Abs were observed in H-2(a/b) mice immunized with the peptide vaccines after the challenge with Friend virus, compared with the responses of the control mice. Detailed kinetic and immunohistopathologic analyses suggested that Th cells might be directly involved in the growth inhibition and elimination of virus-infected erythroid precursor cells.
  • K MURAKAMI; T ABE; M MIYAZAWA; M YAMAGUCHI; T MASUDA; T MATSUURA; S NAGAMORI; K TAKEUCHI; K ABE; M KYOGOKU
    LABORATORY INVESTIGATION 72 6 731 - 739 1995年06月 [査読有り]
     
    BACKGROUND: Thus far, human hepatic Ito (fat storing) cell lines have not been established. Therefore, functional characteristics of Ito cells have not been fully investigated. EXPERIMENTAL DESIGN: We established a new cell line, LI90, that exhibited characteristics compatible with those of Ito cells from a human hepatic mesenchymal tumor. LI90 cells were examined with phase-contrast microscopy, immunohistochemistry, and cytogenetics, and their vitamin A-storing activity was analyzed. To obtain a marker specific for Ito cells for immunohistochemical analyses, we raised mAb against LI90 cells and clarified the molecular nature of the Ag recognized with the new Ab using an expression cloning approach. RESULTS: LI90 cells showed polygonal shape and had well developed cu-smooth muscle actin filaments in their cytoplasm. In an overconfluent culture condition, LI90 cells aggregated to form a typical hills-and-valleys structure. LI90 cells produced various connective tissue components, such as collagen types I, III, IV, V, and VI, laminin, and fibronectin. In culture media containing vitamin A, LI90 cells formed many fat droplets in their cytoplasm, and fluorescence characteristic of vitamin A was observed in the droplets. By immunizing mice with LI90 cells, three separate mAb specifically reacting with Ito cells in human liver sections were established, and the Ag recognized with all three Ab were identified as extracellular matrix tenascin. CONCLUSIONS: The above-described morphologic and functional characteristics, including vitamin A-storage and biosynthesis of tenascin, are compatible with those of Ito cells. Therefore, LI90 cells will be useful for in vitro studies of functions of human Ito cells.
  • LL PERRY; M MIYAZAWA; K HASENKRUG; K WEHRLY; CS DAVID; B CHESEBRO
    JOURNAL OF VIROLOGY 68 8 4921 - 4926 1994年08月 [査読有り]
     
    Resistance to erythroleukemia induced by infection with the Friend virus complex (FV) has been mapped to several genes residing both within and outside the murine major histocompatibility complex (MHC). MHC genes located in the A, D, and Qa/Tla regions of the murine H-2 complex have been shown to affect disease resistance through their capacity to regulate various aspects of the host immune response to viral antigens. This study establishes H-2E as the fourth MHC locus controlling immunological resistance to FV. Our investigation into the role of H-2E molecules revealed two distinct and opposite effects on recovery from Friend disease. H-2(b/b) mice normally lack a functional E gene product and are resistant to high doses of FV. The expression of H-2E molecules in H-2 recombinant or transgenic mice of this genotype resulted in a significant decrease in spontaneous recovery from FV-induced leukemia. In contrast, H-2E expression also appeared to influence recovery from Friend disease in a positive manner, since blocking these molecules with anti-E antibodies in vivo significantly decreased recovery from Friend disease. The data indicate that the positive effects of H-2E molecules derive from their function as restriction elements for helper T-cell recognition of the viral envelope glycoprotein, and eve postulate that the negative effects are due to H-2E-dependent deletions in the T-cell repertoire during development.
  • S MORI; M NOSE; M MIYAZAWA; M KYOGOKU; JB WOLFINBARGER; ME BLOOM
    AMERICAN JOURNAL OF PATHOLOGY 144 6 1326 - 1333 1994年06月 [査読有り]
     
    Aleutian mink disease (AD) has been characterized by immune complex glomerulonephritis associated with persistent infection of Aleutian mink disease parvovirus (ADV). Histopathological examination of kidneys from ADV-infected mink in this study revealed that interstitial nephritis characterized by prominent damage of renal tubuli and lymphocyte infiltration was also common in AD along with glomerulonelphritis. By using strand-specific in situ molecular hybridization technique, replication of ADV was observed in tubular eipithelial cells, in addition to epithelial cells of Bowman's capsules and some glomerular cells of the infected mink. Analysis of tubular lesions by a combination of immunohistochemistry and in situ hybridization revealed that the renal tubili positive for virion DNA or replicative form DNA/mRNA of ADV were also positive for an activation marker ofimmunocompetent cells, which is shared by B lymphocytes and thymic epithelial cells. Infiltration of a subpopulation of T lymphocytes around infected renal tubuli were observed but deposition of immune complexes in these tubular lesions was not demonstrable ADV replication in epithelial cells of renal tubuli and cell mediated immune responses to the infected epithelial cells may play a role in the pathogenesis of interstitial nephritis in Aleutian mink disease.
  • M MIYAZAWA; R FUJISAWA
    TOHOKU JOURNAL OF EXPERIMENTAL MEDICINE 173 1 91 - 103 1994年05月 
    Spontaneous and induced immunity against exogenous Friend murine leukemia retrovirus infection is controlled by four H-a-linked host genes and a single non-H-a gene. Recognition of the envelope glycoprotein gp70 of Friend virus by helper T cells is restricted by A(b) and hybrid E(b/k(d)) class II MHC molecules. By expressing portions of the enu gene and by utilizing synthetic oligopeptides, an A(b)-restricted and a hybrid E(b/d)-restricted helper T cell epitopes were identified in the N- and C-terminal portions of gp70, respectively. These epitopes differ in their amino acid sequences from the previously reported endogenous retroviral peptides naturally presented by mouse MHC class II molecules. Possible roles of recombinant polytropic viruses in the induction of autoimmune responses against endogenous retroviral antigens are also discussed.
  • M MIYAZAWA; S MORI; GJ SPANGRUDE; JB WOLFINBARGER; ME BLOOM
    HYBRIDOMA 13 2 107 - 114 1994年04月 [査読有り]
     
    Several hybridoma clones that produce monoclonal antibodies (MAbs) reacting with subpopulations of mink lymphoid cells were established. Two of the MAbs, MTS-4.3 and MTS-9.3, reacted with relatively small populations of surface immunoglobulin (Ig)-negative (Ig(-)) lymphocytes. MTS-4.3(+) and MTS-9.3(+) cells were distributed in the thymic cortex and medulla, paracortical areas of lymph nodes, and periarterial lymphoid sheaths of the spleen, indicating that these MAbs identify T lymphocytes. Another MAb, MTB-5.6, reacted with a large proportion of surface Ig(+) lymph node cells, but not with surface Ig(-) cells. In immunohistochemistry this MAb stained dendritic epithelial cells of thymic cortex, large polygonal cells of thymic medulla, a large proportion of lymphocytes in the mantle zone of lymphoid follicles, dendritic-shaped cells of paracortical area, and some lymphocytes and macrophage-like cells of medullary cords and sinuses of lymph nodes. The expression of the cell-surface antigen reacting with MTB-5.6 on Ig(+) lymph node cells was increased after concanavalin A stimulation. These new reagents may be useful to analyze cellular basis of the abnormal immune responses observed in Aleutian mink disease, a classical model of human autoimmune diseases.
  • H KANNO; M NOSE; T NIKI; M MIYAZAWA; M KYOGOKU
    TOHOKU JOURNAL OF EXPERIMENTAL MEDICINE 171 1 43 - 52 1993年09月 [査読有り]
     
    Human T cell leukemia virus type I-transformed T cell line HUT102 constitutively secreted soluble factors which induced differentiation of a murine myeloid leukemic cell line, M1, to increase the immune complex-binding and/or phagocytizing capacity. This macrophage differentiating factor(s) (MDF) was purified from the culture supernatants of HUT102 cells by using several steps of column chromatography and novel immune-adherence and/or immune-phagocytic assays. The finally purified MDF activity was detected in the fraction that consisted of 40,000- and 45,000- molecular weight molecules. Antibodies specific for human interleukin-6 or for human granulocyte-colony stimulating factor, both of which have differentiation-inducing activity on M1 cells when used as a single factor, could not neutralize the MDF activity. These findings suggest that the 40,000- and/or 45,000- molecular weight molecules in the HUT102 cell products may be possible novel differentiation-inducing factors acting on a murine macrophage lineage across the species barrier.
  • M IWASHIRO; T KONDO; T SHIMIZU; H YAMAGISHI; K TAKAHASHI; Y MATSUBAYASHI; T MASUDA; A OTAKA; N FUJII; A ISHIMOTO; M MIYAZAWA; MN ROBERTSON; B CHESEBRO; K KURIBAYASHI
    JOURNAL OF VIROLOGY 67 8 4533 - 4542 1993年08月 [査読有り]
     
    To identify retroviral antigenic determinants recognized by CD4+ T helper cells during tumor rejection, we established four noncytolytic, helper-type, CD4+ T-cell clones by limiting dilution cultures of mixed lymphocyte-tumor cultures from mice immune to a Friend virus-induced tumor, FBL-3. Among these, three T helper cell clones were isolated from C57BL/6 mice and the fourth was isolated from a (BALB/c x C57BL/6)F1 mouse. All these clones proliferated in response to the immunizing FBL-3 tumor cells in a major histocompatibility complex class II-restricted manner. Each clone expressed a distinct T-cell receptor with a characteristic combination of alpha and beta chains. The localization of helper T-cell determinants on viral proteins was analyzed with recombinant vaccinia viruses expressing Friend murine leukemia virus (F-MuLV) gag or env genes or shorter fragments of the env gene. Epitopes recognized by these T-cell clones were mapped to at least two distinct portions in the env region of the F-MuLV genome. These epitopes were identified more precisely with synthetic peptides derived from the F-MuLV envelope protein sequence. One of these epitopes was common to Friend and Moloney MuLVs and was located in the N-terminal region of the gp70 glycoprotein at amino acids 122 to 141. The second epitope, which was recognized in the context of hybrid I-E(b/d) major histocompatibility complex class II molecule, was located close to the C-terminal end of gp70 at amino acids 462 to 479. In addition, a possible third epitope was located in the N-terminal half of the gp70 sequence and differed from the first epitope in that it was not cross-reactive with the Moloney MuLV envelope protein,
  • Y IWASAKI; K SAKO; Y OHARA; M MIYAZAWA; M MINEGISHI; S TSUCHIYA; T KONNO
    ACTA NEUROPATHOLOGICA 85 5 566 - 572 1993年04月 [査読有り]
     
    A unique form of subacute panencephalitis developed in a child with aplastic anemia 8 months after an allogeneic bone marrow transplantation (BMT). It was characterized by parenchymal infiltration of CD3 lymphocytes, a marked increase in the number of microglia strongly expressing HLA-DR antigens in both the gray and white matter, and diffuse degeneration of the cerebral white matter. The onset of neurological symptoms coincided with the development of chronic systemic graft-versus-host disease (GVHD). Cellular infiltrates in the CNS lesions were exclusively CD3 lymphocytes intermingled with a small number of monocytes labeled with CD68. There was a preponderance of cells of the CD45RB phenotype. The pathological changes in visceral organs were consistent with those of chronic GVHD. In addition. scrutiny of immunohistochemistry disclosed sparse infiltration of CD3 lymphocytes and diffuse gliosis in the cerebral white matter of another child with chronic GVHD who died 9 months after allogeneic BMT. These cases are suggestive of a potential risk of CNS involvement in GVHD.
  • 呉 敏; 宮沢 正顕; 能勢 真人
    Drug Delivery System 8 1 5 - 10 日本DDS学会 1993年 
    Immunotoxins (ITs) are antibody-based reagents which generally consist of monoclonal antibodies(mAbs) and toxins or their subunits. For the last two decades studies of ITs have made great progress in treating cancers. More recently, possibilities of treating AIDS and some autoimmune diseases by ITs have stared to attract attentions of researchers who are looking for effective drugs for these intractable diseases. Previously constructed ITs for infectious diseases were mainly for HIV ; however, ITs for other viral infection are also valuable to be investigated. The only ITs so far studied for the treatment of autoimmune diseases are those specific for IL-2, IL-2 receptor, and L3T4. In this report, we discuss possible use of ITs for treatment of autoimmune diseases and viral infections referring to the latest related publications.
  • T MIZOI; H OHTANI; K MIYAZONO; M MIYAZAWA; S MATSUNO; H NAGURA
    CANCER RESEARCH 53 1 183 - 190 1993年01月 [査読有り]
     
    Transforming growth factor beta1 (TGF-beta1) is secreted as an inactive complex associated with latent TGF-beta1 binding protein (LTBP). Tissue localization of these proteins has not been fully understood in human pathological conditions. We examined the immunohistochemical localization of TGF-beta1 precursor (proTGF-beta1) and LTBP in carcinomas and granulation tissue in the human gastrointestinal tract at the light and electron microscopic levels. In normal tissue, endothelial cells and granulocytes sporadically showed immunoreactivity for proTGF-beta1, while epithelial cells were all negative. In cancer tissue, both cancer cells and stromal cells (fibroblasts, macrophages, and endothelial cells) were positive for proTGF-beta1, more frequently in diffuse-type gastric carcinomas than in differentiated-type adenocarcinomas. Immunoelectron microscopy revealed that proTGF-beta1 was localized in rough endoplasmic reticulum and perinuclear cisternae in fibroblasts, macrophages, and endothelial cells in cancer stroma and in fibrous granulation tissue. In contrast, the intracellular localization of proTGF-beta1 in carcinoma cells was predominantly observed in the cytosol (cytoplasmic matrix). This finding suggests disarranged or blocked intracellular transportation of proTGF-beta1 in cancer cells. The immunoreactivity for LTBP was not observed in the normal epithelial cells. It was localized in cancer stroma, not in cancer cells. Ultrastructurally, LTBP was located in the extracellular matrix around fibroblasts and smooth muscle cells. The intracellular immunoreactivity for LTBP was observed in rough endoplasmic reticulum of fibroblasts and smooth muscle cells, the same as in granulation tissue. These results suggest that gastrointestinal carcinoma cells produce no or a small amount of LTBP in vivo. Our investigation suggests that extensive fibrosis in both cancer stroma and granulation tissues may be promoted by TGF-beta1 mainly secreted from stromal cells.
  • M MIYAZAWA; J NISHIO; B CHESEBRO
    JOURNAL OF VIROLOGY 66 7 4497 - 4507 1992年07月 [査読有り]
     
    High sequence variability in the envelope gene of human immunodeficiency virus has provoked interest in nonenvelope antigens as potential immunogens against retrovirus infection. However, the role of core protein antigens encoded by the gag gene in protective immunity against retroviruses is unclear. By using recombinant vaccinia viruses expressing the Friend murine leukemia helper virus (F-MuLV) gag gene, we could prime CD4+ T-helper cells and protectively immunize susceptible strains of mice against Friend retrovirus infection. Recovery from leukemic splenomegaly developed more slowly after immunization with vaccinia virus-F-MuLV gag than with vaccinia virus-F-MuLV env; however, genetic nonresponders to the envelope protein could be partially protected with Gag vaccines. Class switching of F-MuLV-neutralizing antibodies from immunoglobulin M to immunoglobulin G after challenge with Friend virus complex was facilitated in mice immunized with the Gag antigen. Sequential deletion of the gag gene revealed that the major protective epitope was located on the N-terminal hydrophobic protein p15.
  • M MIYAZAWA; J NISHIO; K WEHRLY; G JAY; RW MELVOLD; B CHESEBRO
    EUROPEAN JOURNAL OF IMMUNOGENETICS 19 3 159 - 164 1992年06月 [査読有り]
     
    Using H-2 recombinant and mutant mice, the Rfv-1 gene influencing spontaneous recovery from Friend retrovirus (FV)-induced leukaemia was mapped in the D locus. Two D(b) alleles were required for full recovery, and a single D(d) transgene did not convey increased susceptibility to FV in the presence of homozygous D(b/b) genotype. The results suggest that an increase in the expression of D(b) may lead to more effective stimulation of FV-specific CTL.
  • M MIYAZAWA; J NISHIO; K WEHRLY; CS DAVID; B CHESEBRO
    JOURNAL OF IMMUNOLOGY 148 6 1964 - 1967 1992年03月 [査読有り]
     
    The Rfv-2 gene that influences the rate of spontaneous recovery from erythroleukemia induced by a low dose of Friend retrovirus complex was mapped to the Q/TL region of mouse MHC. Rfv-2 was physically and functionally distinct from the I-A-linked Ir gene that has been shown to control the responsiveness of Th cells to the envelope glycoprotein of Friend murine leukemia helper virus. The negative effect of the Rfv-2s allele was overcome by the B10.D2-H-2dm1 mutation of the D-L genes of H-2, suggesting functional similarities between the D-L and Q/TL genes in influencing resistance against Friend murine leukemia retrovirus complex infection or possible modification of Q/TL expression by genes in the D-L region.
  • M SUGAMATA; M MIYAZAWA; S MORI; GJ SPANGRUDE; LC EWALT; DL LODMELL
    JOURNAL OF VIROLOGY 66 2 1252 - 1260 1992年02月 [査読有り]
     
    Street rabies virus (SRV)-infected T-lymphocyte-deficient (nude) mice, in contrast to euthymic mice, did not develop hindlimb paralysis prior to death. To document the role of T lymphocytes in rabies virus-associated paralysis, 10(8) spleen cells from normal immunocompetent euthymic mice were transferred to nude mice and the recipient mice were challenged with SRV. One hundred percent of the reconstituted mice developed paralysis and died. Depletion of T cells from the donor spleen suspension prior to transfer abrogated the development of paralysis but did not prevent the deaths of the recipient animals. Mice receiving 10(8) rabies virus-immune spleen cells did not become paralyzed and did not die. Nude mice inoculated with either rabies virus-immune or normal mouse serum prior to and following SRV inoculation did not develop paralysis. Immune serum protected the mice, whereas animals inoculated with normal serum died. Central nervous system inflammatory responses in nude mice immunologically reconstituted with normal spleen cells were characterized by diffuse cellular infiltrates in the parenchyma and extensive perivascular cuffing. Perivascular infiltrates included CD8+ and CD4+ T lymphocytes and Mac-1+ macrophage-microglial cells. Inflammatory cells in the parenchyma were limited to CD8+ lymphocytes and Mac-1+ cells. These observations indicate that paralysis of SRV-infected mice is dependent on T lymphocytes. Whether injury leading to paralysis is mediated by T lymphocytes or by an influence of T lymphocytes on macrophage-microglial cells or other cells remains to be determined.
  • M MIYAZAWA; J NISHIO; M KYOGOKU; B CHESEBRO
    MOLECULAR APPROACHES TO THE STUDY AND TREATMENT OF HUMAN DISEASES 966 177 - 184 1992年 [査読有り]
  • S MORI; M NOSE; M MIYAZAWA; M KYOGOKU; JB WOLFINBARGER; ME BLOOM
    MOLECULAR APPROACHES TO THE STUDY AND TREATMENT OF HUMAN DISEASES 966 301 - 308 1992年 [査読有り]
  • C ISHIHARA; M MIYAZAWA; J NISHIO; AZUMA, I; B CHESEBRO
    VACCINE 10 5 353 - 356 1992年 [査読有り]
     
    Low toxic and synthetic adjuvants were investigated in the induction of protective immunity against Friend murine retrovirus-induced erythroleukaemia by immunization with inactivated Friend murine leukaemia helper virus (F-MuLV). 6-O-(2-tetradecyl-hexadecanoyl)-N-acetylmuramyl-L-alanyl-D-isoglutamine (B30-MDP) showed a significant enhancement of the protective immunity against Friend virus-induced erythroleukaemia not only in H-2a/b mice known to make good immune responses to F-MuLV envelope, but also in H-2a/a mice which are usually unable to respond to F-MuLV envelope protein. Another analogue of N-acetylmuramyl-D-isoglutamine (MDP), N(alpha)-acetylmuramyl-L-alanyl-D-isoglutaminyl-N(epsilon)-stearoyl-L-lysine [MDP-Lys(L18)], which has been shown to enhance non-specific protective activity against bacterial and viral infections, however, showed no adjuvant activity in the present system. A combined adjuvant of the synthesized mycobacterial cord factor, trehalose dimycolate (TDM) and detoxified bacterial endotoxin, monophosphoryl lipid A from Salmonella minnesota, gave good protection which was comparable to complete Freund's adjuvant in both H-2a/b and H-2a/a mice.
  • M MIYAZAWA; J NISHIO; K WEHRLY; B CHESEBRO
    JOURNAL OF IMMUNOLOGY 148 2 644 - 647 1992年01月 [査読有り]
     
    Genes influencing the rate of spontaneous recovery from erythroleukemia induced by a low dose of Friend virus complex were located in the right and left portions of the mouse MHC. The right side gene was most likely the previously described Rfv-1 in the H-2D region. Using the B6.C-H-2bm12 mutant mice, the left side gene was mapped to the A-beta class II locus. The A-beta(b) was a resistant allele and A-beta(k) and A-beta(bm12) were susceptible alleles. Genes at this class II locus controlled the responsiveness of Th cells to envelope glycoprotein of Friend murine leukemia helper virus and affected the class switching of virus-neutralizing antibodies from Igm to IgG in FV-infected mice.
  • MN ROBERTSON; M MIYAZAWA; S MORI; B CAUGHEY; LH EVANS; SF HAYES; B CHESEBRO
    JOURNAL OF VIROLOGICAL METHODS 34 3 255 - 271 1991年10月 [査読有り]
     
    Four monoclonal antibodies were selected for their ability to recognize the envelope protein of Friend murine leukemia virus (F-MuLV) in methanol-fixed tissue culture cells. Each of these monoclonal antibodies was found to react only with F-MuLV. By using recombinant retroviruses, it was determined that each of the monoclonal antibodies recognized the C-terminal one-third of the F-MuLV gp70 envelope protein. The monoclonal antibodies were effective in radioimmunoprecipitation of F-MuLV proteins, and one of the antibodies, 720, was also effective in Western blotting. The ability of antibody 720 to react with F-MuLV in methanol-fixed cells facilitated the use of a sensitive immunoperoxidase method with a focal virus infectivity assay. In immunohistochemical studies using light microscopy, antibody 720 could specifically label F-MuLV-infected cells in acetone-fixed tissue sections from F-MuLV-infected animals. Finally, in immuno-gold labelling studies using electron microscopy, antibody 720 could be used to distinguish F-MuLV from amphotropic MuLV.
  • C ISHIHARA; M MIYAZAWA; J NISHIO; B CHESEBRO
    JOURNAL OF IMMUNOLOGY 146 11 3958 - 3963 1991年06月 [査読有り]
     
    (B10.A x A/WySn)F1, H-2a/a, mice are genetic nonresponders to the envelope protein of Friend murine leukemia helper virus (F-MuLV) when immunized with a recombinant vaccinia virus expressing F-MuLV env gene. In contrast these mice can be protectively immunized against leukemogenic Friend virus complex using formalin-fixed F-MuLV virions in CFA. To determine which viral proteins were responsible for this immune protection, virion proteins prepared by SDS-PAGE and electroelution were used to immunize mice. Purified gp70 envelope protein in CFA was capable of inducing strong immune protection against the challenge with Friend virus complex in H-2a/a mice. Immunologic studies demonstrated that immunized mice developed a virus-specific T cell proliferative response and showed IgM to IgG Ig class switching of virus-neutralizing antibodies. These results indicated that genetically controlled immune nonresponsiveness to F-MuLV envelope Ag in H-2a/a mice could be overcome using denatured viral envelope protein together with a strong adjuvant.
  • S MORI; JB WOLFINBARGER; M MIYAZAWA; ME BLOOM
    JOURNAL OF VIROLOGY 65 2 952 - 956 1991年02月 [査読有り]
     
    By using strand-specific in situ hybridization and immunohistochemistry, evidence for replication of the Aleutian mink disease parvovirus was observed in cells resembling macrophages and cells resembling follicular dendritic cells at 10 days after infection but only in macrophages at 60 days. Sequenstration of the Aleutian mink disease parvovirus in larger numbers of macrophages and follicular dendritic cells was noted at both 10 and 60 days.
  • CHESEBRO B.
    Annu. Rev. Immunol. 8 1 477 - 499 1990年04月
  • M MIYAZAWA; J NISHIO; B CHESEBRO
    VACCINES 90 407 - 412 1990年 [査読有り]
  • S YOU; S MAEDA; S MURAO; R TAKAHASHI; J ISHIKAWA; M MIYAZAWA; M NOSE; T SUGIYAMA
    JAPANESE JOURNAL OF CANCER RESEARCH 80 5 444 - 451 1989年05月 [査読有り]
  • M MIYAZAWA; J NISHIO; B CHESEBRO
    JOURNAL OF EXPERIMENTAL MEDICINE 168 5 1587 - 1605 1988年11月 [査読有り]
  • M MIYAZAWA; M NOSE; M KAWASHIMA; M KYOGOKU
    JOURNAL OF EXPERIMENTAL MEDICINE 166 4 890 - 908 1987年10月 [査読有り]
  • Kyogoku M; Nose M; Sawai T; Miyazawa M; Tachiwaki O; Kawashima M
    Progress in clinical and biological research 229 95 - 130 1987年 [査読有り]
  • Nose M; Miyazawa M; Tachiwaki O; Kyogoku M
    Nihon rinsho. Japanese journal of clinical medicine 43 10 2044 - 2051 1985年10月 [査読有り]

書籍

  • 近藤, えり; 宮澤, 正顯 チャイルド社 2020年09月 ISBN: 9784925258524 31p
  • 解明病理学 病気のメカニズムを解く
    医歯薬出版 2009年 ISBN: 9784263731154
  • 述２
    明石書店 2008年 ISBN: 9784750327495
  • Host genes that influence immune and non-immune resistance mechanisms against retrovrial infections
    Recent Research Developements in Virology 2005年
  • Host genes that influence immune and non-immune resistance mechanisms against retrovrial infections
    Recent Research Developements in Virology 2005年
  • 菅原, 努; 宮澤, 正顯; 大東, 肇 昭和堂 2004年04月 ISBN: 4812204011 v, 144p
  • Pathogenicity of autoantibodies reactive with the endogenous retroviral envelope glycoprotein gp70 (In "From Animal Models to Human Genetics: Research on the Induction and Pathogenicity of Autoantibodies
    Pabst Science Publishers 2004年
  • 山岸, 秀夫; 宮澤, 正顯 昭和堂 2001年10月 ISBN: 481220125X xv, 168, iiip
  • Tumor Imaging
    Encyclopedia of Immunology, 2nd edition 1998年

講演・口頭発表等

  • HIV感染移行性の分子機構： Rac2とAPOBEC3  [通常講演]
    第24回日本エイズ学会学術集会・総会 2010年
  • 性感染症の最近の動向： HIVと性器クラミジアを中心に （教育講演）  [通常講演]
    第19回日本口腔感染症学会総会・学術大会 2010年
  • レトロウイルス遺伝子発現と糸球体病変： 拡大するヒトレトロウイルスの世界 （特別講演）  [通常講演]
    第45回日本小児腎臓病学会学術集会 2010年
  • Genetic factors that confer resistance to HIV-1 acquisition in HIV-1-exposed but seronegative individuals in Italy and Thailand.  [通常講演]
    4th Nagasaki Symposium on Tropical and Emerging Infectious Diseases. 2009年
  • New metabolic markers derived from gamma and deltaretrovirus envelope glycoproteins.  [通常講演]
    21st International Workshop on Retroviral Pathogenesis. 2009年
  • Mechanisms of immune evasion by Friend virus: T-cell exhaustion, B-cell hyperactivation, and their genetic control.  [通常講演]
    21st International Workshop on Retroviral Pathogenesis. 2009年
  • Mouse APOBEC3 affects the production of virus-neutralizing antibodies by restricting early retroviral replication, not by altering the B-cell repertoire.  [通常講演]
    Frontiers of Retrovirology. 2009年
  • New metabolic markers derived from gamma and deltaretrovirus envelope glycoproteins.  [通常講演]
    21st International Workshop on Retroviral Pathogenesis 2009年
  • Mechanisms of immune evasion by Friend virus: T-cell exhaustion, B-cell hyperactivation, and their genetic control.  [通常講演]
    21st International Workshop on Retroviral Pathogenesis 2009年
  • Mouse APOBEC3 affects the production of virus-neutralizing antibodies by restricting early retroviral replication, not by altering the B-cell repertoire.  [通常講演]
    Frontiers of Retrovirology 2009年
  • Host genes controlling immune responses to retroviral infections and genetic correlates of HIV-1-reposed but uninfected status.  [通常講演]
    1st International Symposium on Genetic and Immune Correlates of HIV Infection and Vaccine-Induced Immunity. 2007年
  • HIV-1感染抵抗性を賦与する新規宿主遺伝子の解析とその作用機序  [通常講演]
    第２１回日本エイズ学会学術集会・総会 2007年
  • A cluster of retrovirus-restricting genes in mouse chromosome 15 and syntenic human chromosome 22 and their effects on Friend virus infection.  [通常講演]
    19th International Workshop on Retroviral Pathogenesis 2007年
  • Host genes controlling immune responses to retroviral infections and genetic correlates of HIV-1-reposed but uninfected status.  [通常講演]
    1st International Symposium on Genetic and Immune Correlates of HIV Infection and Vaccine-Induced Immunity. 2007年
  • A cluster of retrovirus-restricting genes in mouse chromosome 15 and syntenic human chromosome 22 and their effects on Friend virus infection.  [通常講演]
    19th International Workshop on Retroviral Pathogenesis 2007年
  • Host genes controlling immune responses to retroviral infections and genetic correlates of HIV-1-reposed but uninfected status.  [通常講演]
    1st International Symposium on Genetic and Immune Correlates of HIV Infection and Vaccine-Induced Immunity. 2007年
  • ビルマ産アカゲザルMHC class II領域のハプロタイプ構成と組換えの解析  [通常講演]
    ビルマ産アカゲザルMHC class II領域のハプロタイプ構成と組換えの解析 2006年
  • Host genetic factors that control immune resistance to HIV-1 infection.  [通常講演]
    7th AIDS Seminar in Kumamoto 2006年
  • Host genetic factors that control immune resistance to HIV-1 infection.  [通常講演]
    7th AIDS Seminar in Kumamoto 2006年
  • Host resistence genes in immunity against human and mouse retroviral infections.  [通常講演]
    Virology Africa 2005 2005年
  • Host resistence genes in immunity against human and mouse retroviral infections.  [通常講演]
    Virology Africa 2005 2005年
  • Genetic basis for resistance against retroviral infections − from mouse models to humans.  [通常講演]
    Third International Workshop on Immunology and Infectious Diseases 2004年
  • Genetic basis for resistance against retroviral infections − from mouse models to humans.  [通常講演]
    Third International Workshop on Immunology and Infectious Diseases 2004年

作品等

  • 温熱によるC-型肝炎ウイルス免疫応答の増強
    2001年 -2007年
  • Enhancement of immune responses against HCV by heat treatment(hyperthermia)
    2001年 -2007年
  • 新型インフルエンザワクチンの作用機序
    2006年
  • ヘルパーT細胞を活性化するエイズワクチンの開発
    2000年 -2005年
  • Development of helper T-cell vaccines against HIV infection
    2000年 -2005年
  • HIV曝露非感染者の遺伝要因の解明
    2002年
  • Molecular identification of host genetic factors determining HIV-exposed but uninfected status
    2002年
  • 乳酸菌製剤の抗ウイルス免疫賦活能
    1999年 -2000年
  • Induction of antiviral immune responses by using Lactobacillus products
    1999年 -2000年
  • エイズ感染防御を含む新しいワクチンへの現代的アプローチに関するコールドスプリングハーバー研究所シンポジウムシンポジスト
    1989年
  • Cold Spring Harbor Symposium on Modern Approaches to New Vaccines Including Prevention of AIDS
    1989年
  • ワクチン開発のための新しい化学的・遺伝的アプローチに関するコールドスプリングハーバー研究所シンポジウムシンポジスト
    1987年
  • Cold Spring Harbor Symposium on New Chemical and Genetic Approaches to Vaccination
    1987年

MISC

産業財産権

  • Marker Genes
    PCT/GB2003/004493
  • 新規ヒト内在性レトロウイルスHC2のenv遺伝子
    特願2004-231412
  • Resistance Genes
    International Patent Application PCT/GB2005/005078
  • Method for diagnosis and induction of resistance to virus.
    International Patent Application PCT/JP2007/068591
  • Marker Genes
    PCT/GB2003/004493
  • Resistance Genes
    International Patent Application PCT/GB2005/005078
  • Method for diagnosis and induction of resistance to virus.
    International Patent Application PCT/JP2007/068591

受賞

  • 2007年 内藤記念科学奨励金
     JPN
  • 2002年 ノバルティス リウマチ医学賞
     JPN
  • 2002年 Novartis Rheumatism Research Award
  • 1990年 難病医学研究財団研究奨励助成金
     JPN
  • 1989年 東北大学医学部奨学賞
     JPN

共同研究・競争的資金等の研究課題

  • レトロウイルス中和抗体産生制御遺伝子の実体と作用機序の解明
    文部科学省：科学研究費補助金（基盤研究（B））
    研究期間 : 2019年04月 -2022年03月 
    代表者 : 宮澤 正顯
  • マウスAPOBEC3のプロテアーゼ抑制機構解明による新規抗レトロウイルス薬開発
    文部科学省：科学研究費補助金（基盤研究（B））
    研究期間 : 2016年04月 -2019年03月 
    代表者 : 宮澤 正顯
  • 細胞内複製制限因子ＡＰＯＢＥＣ３の進化要因としての異種由来レトロウイルス
    文部科学省：科学研究費補助金 新学術領域研究（研究領域提案型）
    研究期間 : 2015年04月 -2017年03月 
    代表者 : 宮澤 正顯
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2012年 -2014年 
    代表者 : 宮澤 正顯
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2009年 -2011年 
    代表者 : 宮澤 正顯; 河原 佐智代; 博多 義之; 高村 史記
     
    マウスAPOBEC3遺伝子には機能的多型があり、レトロウイルス感染に自然抵抗性の系統では造血系組織、特にBリンパ球で遺伝子発現が高く、その転写産物は第5エキソンを欠くものが主体を占める。APOBEC3遺伝子多型はそのタンパク質発現量にも影響するが、これは第5エキソンの有無が翻訳効率を決定するためである。第5エキソン取り込みの有無を決めるのは、第4イントロンのRNA分岐部位多型と第5エキソン内の単一塩基多型であることを解明した。
  • 宿主の自然抵抗性を応用した抗HIV/エイズ戦略の開発
    厚生労働科学研究費補助金
    研究期間 : 2008年
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2005年 -2006年 
    代表者 : 宮澤 正顕; 金成 安慶
     
    ヒト第22染色体に存在する、HIV曝露非感染状態を決定する遺伝子について、研究期間内にその最終候補をほぼ絞り込むことに成功した。即ち、DNAマイクロアレイを用いた発現解析によって、末梢血単核球のHIV抗原刺激に伴い、HIV曝露非感染者では発現が上昇し、感染者では逆に発現が低下する遺伝子二つを見出したが、これら二遺伝子は染色体上で互いに隣接して、しかも逆向きに存在していた。一方、同じ染色体領域の単一塩基多型(SNPs)の遺伝子型を群間で比較したところ、上記二遺伝子にごく隣接した二つの短い染色体領域で、曝露非感染者群とHIV感染者群の間に有意な頻度差を認めた。そこで、曝露非感染者で発現上昇が見られる二つの遺伝子の発現調節領域を探ったところ、これら二つの遺伝子の構造領域の間に、リプレッサーと思われる配列を見出した。しかも、このリプレッサー候補領域にはゲノム多型があって、互いにハプロタイプ関係をなす特定の遺伝子型が、曝露非感染者に集積していた。さらに、試験管内のルシフェラーゼ発現誘導実験でも、これらゲノム塩基配列の多型が遺伝子発現に影響することが明らかになった。さらに、上記ゲノム塩基配列多型領域の僅かにテロメア寄りにある、レトロウイルス複製制御因子APOBEC3Gについて、その末梢血単核球での発現が曝露非感染者で有意に高く、特にCD14陽性単球を1型インターフェロンで刺激した場合のAPOBEC3G発現誘導が、曝露非感染者で有意に高度となることが明らかとなった。このことから、単球におけるAPOBEC3Gの高発現が、曝露非感染者のHIV感染抵抗性の一機序であることが考えられる。今後、上記ゲノム塩基配列多型とAPOBEC3G発現誘導との関係が明らかにされねばならない。
  • 文部科学省：科学研究費補助金(基盤研究(B))
    研究期間 : 2003年 -2004年 
    代表者 : 宮澤 正顯; 阿部 弘之; 金成 安慶
     
    イタリアコホートのHIV-1曝露非感染者42名、それらのHIV-1感染パートナー42名と同一地区・年齢層の他の感染者7名、及び同一地区・年齢層の非感染健常者47名を対象に、第22染色体マーカーの遺伝子型を決定した。その結果、D22S277遺伝子座について、対立遺伝子156または158を持つ者の頻度が、曝露非感染者群でHIV-1感染者群或いは健常者群のおよそ4倍高く、多重比較補正後も有意差があること(p=0.038-0.045)、D22S423についても、対立遺伝子229の頻度が曝露非感染者群で感染者群の4倍以上高く、多重比較補正後も有意差があること(p=0.032)が示された。また、感染者および健常者群ではD22S284からD22S1166に至る連鎖不平衡が認められるが、曝露非感染者群では、この領域の連鎖不平衡がD22S276座で分断されていた。そこで、上記候補遺伝子存在範囲を含むMb遺伝子からPacsin2遺伝子までの領域について、そこに含まれる全既知遺伝子、および機能未同定のopen reading framesを網羅したDNAマイクロアレイを作製した。イタリアコホートの曝露非感染者及び感染者の末梢血単核球を、HIV-1gag及びenv抗原ペプチド混合物で刺激前後RNAを抽出し、発現解析を行ったところ、少数の遺伝子について、曝露非感染者でのみその発現が刺激6時間後に増加していた。また、曝露非感染者でHIV抗原刺激により発現が誘導される遺伝子の内部、または近傍に存在する単一塩基多型(SNPs)について、それらの頻度をイタリア及びタイ・ランパンコホートで解析した。その結果、少なくともイタリアコホートの曝露非感染者で、一候補遺伝子内のSNPとD22S423の遺伝子型に強い連鎖が見られたが、感染者や健常人ではそのような連鎖は認められなかった。
  • 文部科学省：科学研究費補助金(萌芽研究)
    研究期間 : 2003年 -2004年 
    代表者 : 宮澤 正顕; 河原 佐智代
     
    ウイルス感染防御においては、CD8陽性細胞傷害性Tリンパ球(CTL)がウイルス産生細胞を破壊し、中和抗体が感染の拡大とウイルス粒子除去に関与すると信じられている。このため、ワクチン開発に当たって、CTLとウイルス中和抗体の誘導が重要な指標とされることが多い。我々は、免疫系の完成した成体マウスへの接種により致死性赤白血病を誘発するフレンドレトロウイルス(FV)を用い、FVに感受性の高い(BALB/c × C57BL/6)F_1マウスに、β2-ミクログロブリン遺伝子欠損をホモ接合で導入して、CD8陽性細胞を欠く系統を育成、これと野生型F_1マウス、及びBリンパ球を欠く免疫グロブリン遺伝子膜貫通エクソン欠損マウスを用いて、ペプチドワクチンによる感染防御実験を行った。ウイルス被膜タンパク質上のCD4陽性Tリンパ球認識エピトープを単独で含む合成ペプチドで、一度だけ免疫することにまり、野生型マウスの80%以上でFV誘発白血病が予防できた。驚くべきことに、CD8陽性Tリンパ球欠損F_1マウスでも、その約70%がペプチドワクチン免疫によりFVに抵抗性となり、感染後の脾腫発症・死亡率は、免疫した野生型マウスと有意差がなかった。しかし、Bリンパ球欠損マウスでは、ペプチドワクチンで白血病死を防止できなかった。ペプチド免疫マウスでは、CD8陽性Tリンパ球非存在下でも、FV感染4週間後までに骨髄および脾からウイルス産生細胞が排除された。一方、Bリンパ球欠損マウスでは、感染初期のウイルス産生細胞数増加は抑制されたが、感染2週間目以降はワクチンの効果が見られなかった。以上から、CD4陽性Tリンパ球認識抗原エピトープを用いてフレンド白血病ウイルスに対する感染防御を誘導する系に関しては、CD8陽性細胞傷害性Tリンパ球は感染防御に必須ではないことが明らかとなった。
  • 文部科学省：科学研究費補助金(基盤研究(C))
    研究期間 : 2001年 -2002年 
    代表者 : 宮澤 正顯; 田端 信忠; 松村 治雄; 阿部 弘之
     
    1)マウスレトロウイルスgag遺伝子産物MA(マトリクス)タンパク質上に、複数のCD4陽性Tリンパ球認識抗原エピトープを見出した。このうち中央部のアミノ酸残基62番から76番の範囲には、IL-4産生を強く誘導するTh2タイプのエピトープがあり、C-末端の119番から138番の範囲には、IL-4産生を起こさないエピトープがある。2)ペプチドワクチンで免疫したマウスにマウスレトロウイルスを感染させると、エフェクターT細胞の他にナチュラルキラー(NK)細胞が活性化され、NK細胞はレトロウイルス感染細胞を効果的に傷害する。その理由として、レトロウイルス感染細胞ではNK細胞レセプターリガンド分子の発現が上昇していることを見出した。3)MAタンパク質中の感染防御に必須な抗原構造を同定するため、新しいワクシニアウイルスベクターを構築し、MAの部分断片を発現させてマウスを免疫した。その結果、MAのN-末端側を欠失させてもC-末端側を欠失させても感染防御能がなくなった。N-末端側の欠失で感染防御能がなくなる理由として、gag遺伝子産物N-末のミリスチル化が必要である可能性を考え、末端グリシン残基をアラニンに置換したところ、ミリスチル化部位を失ったMAタンパク質は、本来細胞膜の裏打ちに分布するはずのところ、核に移行するようになった。同時に感染防御能も失われたので、MAの全長を発現してもその細胞内分布が変わると感染防御が出来なくなることがわかった。4)N-末のミリスチル化が起こる条件で比較すると、MA中央部のTh2エピトープは感染防御には必要でなく、C-末側のエピトープが必要であることがわかった。
  • HIV曝露非感染状態を決定する宿主遺伝子の同定
    厚生労働科学研究費補助金
    研究期間 : 2001年
  • Identification of novel human genes that determines HIV-exposed, uninfected status
    Health and Labour Sciences Research Grants
    研究期間 : 2001年
  • 文部科学省：科学研究費補助金(基盤研究(C))
    研究期間 : 1999年 -2000年 
    代表者 : 宮澤 正顯; 阿部 弘之; 田端 信忠; 松村 治雄; 宇高 恵子
     
    フレンド白血病ウイルスenv遺伝子産物上に我々が同定したヘルパーT細胞認識エピトープを単独で含む合成ペプチドワクチンは、このウイルスに感受性の高い(BALB/c×C57BL/6)F_1マウスへの一回投与で完全な発症阻止効果を示し、ペプチド投与群ではウイルス接種後早期に感染細胞が排除された。ペプチドワクチンでCD4陽性Tリンパ球を感作した動物で、フレンドウイルス接種後に誘導される感染細胞排除のエフェクター機構を解析するため、免疫マウスにフレンドウイルスを接種後、経時的に脾細胞を採取して細胞傷害試験を行った。その結果、ウイルス感染初期にCD8陽性細胞傷害性Tリンパ球(CTL)が活性化されるほか、ペプチドワクチン投与マウスに限ってウイルス抗原特異的なCD4陽性CTL活性も誘導されること、更に意外なことに、フレンドウイルス接種後7〜9日目に誘導される細胞傷害活性の主体を占めるのは、CD4,CD8が共に陰性の細胞群であることがわかった。このCD4/CD8陰性細胞分画の実体は、マウスNK細胞マーカーDX5を発現する細胞群で、複数のフレンド白血病細胞株及びYAC-1細胞に対し強い傷害活性を示した。更に、ペプチドワクチン免疫マウスより、抗アシアロGM_1抗体投与によってTリンパ球機能に影響を与えずにNK細胞を取り除くと、ワクチンで誘導されたフレンドウイルス感染抵抗性が完全に失われた。一方、ペプチドワクチン投与によるフレンドウイルス感染防御にはCD4陽性及びCD8陽性Tリンパ球の存在も必須であり、特にCD4陽性Tリンパ球の存在は、ウイルス中和抗体産生に欠くことのできないものであった。以上の実験事実から、フレンドウイルス感染の初期にはNK細胞が活性化され、フレンドウイルス感染細胞はNK細胞による傷害に感受性であること、ヘルパーT細胞認識エピトープを単独で含むペプチドワクチンによるフレンドウイルス感染防御には、CD4及びCD8陽性CTLやウイルス中和抗体と共に、NK細胞の機能も必須であることが明らかとなった。
  • 文部科学省：科学研究費補助金(一般研究(C), 基盤研究(C))
    研究期間 : 1995年 -1996年 
    代表者 : 宮澤 正顕; 宮澤 正顕
     
    1.抗gp70抗体移入による糸球体腎炎の発症機構:MRL/Iprマウス由来で、同系正常マウスまたはSCIDマウスへのハイブリドーマ細胞移入により糸球体病変を誘発する抗gp70単クローン抗体を、精製IgG分子としてNZWまたはC57BL/6マウスに移入した。血清gp70値の高いNZWマウスで腎病変の発生を認めたが、C57BL/6マウスでは有意な腎病変は生じなかった。従って、病変発生はgp70-抗gp70免疫複合体の形成を介するものと考えられる。2.組織傷害性抗gp70抗体の認識エピトープ存在部位の絞り込み:レトロウイルスenv遺伝子間のキメラ発現系を用い、腎炎原性抗gp70抗体の反応エピトープの存在部位を限定した。病原性抗gp70抗体の大半は、異種指向性ウイルスgp70分子N-末側に抗原エピトープを持つ。3.内在性レトロウイルスenv遺伝子産物上のヘルパーT細胞認識抗原エピトープの構造解析:既に我々が同定した外来性フレンド白血病ウイルスgp70分子上のヘルパーT細胞認識抗原エピトープ構造を、内在性レトロウイルスenv遺伝子産物の対応部分と比較した。内在性ウイルス由来ペプチドはフレンドウイルス由来抗原ペプチドと12アミノ酸残基中5残基が異なっているが、うち3残基フレンドウイルスと同じものに置換するとT細胞によって確認されるようになった。4.血小板表面への内在性レトロウイルスenv遺伝子産物の発現:抗gp70自己抗体産生ハイブリドーマ細胞の同系正常マウスへの移入による血小板減少性紫班病の発症機構を明らかにするため、MRL/1prマウスと対照の正常マウスから血小板を分離し、内在性レトロウイルスenv遺伝子産物の発現を調べた。殆どのマウスの系統で幾つか異なる種類の内在性レトロウイルスenv遺伝子産物が血小板に発現すること、病原性抗gp70抗体がMRL/1prマウス血小板と直接結合することが明らかとなった。
  • 文部科学省：科学研究費補助金(重点領域研究)
    研究期間 : 1994年 -1994年 
    代表者 : 宮沢 正顕; 宮澤 正顕
     
    HIV感染に対する有効なワクチンの開発には、免疫抑制性のレトロウイルス感染に対する宿主免疫応答の仕組みを基礎研究の面から充分に解析する必要がある。我々は、免疫系の完成した成熟マウスへの接種によって重篤な免疫不全症を伴う急性の赤白血病を誘発するフレンド白血病レトロウイルス複合体をモデルに、レトロウイルス感染に対する宿主免疫応答の抗原特異性と遺伝的制御機構を解析してきた。本年度は、フレンド白血病ヘルパーウイルス(F-MuLV)gag遺伝子の部分発現とminigenesの構築、及び合成オリゴペプチドの作成により、細胞表面に発現するglycosylated Gag蛋白質の前駆体、Pr75^のリーダーペプチド部に、細胞傷害性T細胞(CTL)認識抗原エピトープを同定した。一方、昨年度にF-MuLV env遺伝子産物_70上に同定した二つのヘルパーT細胞認識抗原エピトープを単独で含む合成ペプチドを用い、マウスを免疫することによって、フレンドウイルス複合体に対する感染防御免疫が誘導可能か否かを解析した。その結果_70N-末のエピトープで一回免疫された(C57BL/10×A/J)F_1(H-2^)マウスでは、フレンドウイルス複合体接種後一旦発生した脾腫が感染後40〜60日にかけてゆっくりと退縮して行くこと、一方C-末のエピトープ単独で一回免疫された同じ系統のマウスでは、フレンドウイルス接種後極めて急速に感染細胞の排除が起こり、殆ど脾腫の発生を見ないことが明かとなった。現在、上記ヘルパーT細胞認識エピトープとCTL認識エピトープを共有結合させた多価ペプチドワクチンを作成し、これによって誘導されるレトロウイルス感染防御免疫を解析中である。
  • 文部科学省：科学研究費補助金(重点領域研究)
    研究期間 : 1993年 -1993年 
    代表者 : 宮沢 正顯; 宮澤 正顕
     
    HIV感染者の病勢の進行がウイルス粒子コア部分を形成するGag抗原に対する抗体産生能の消失と相関することは良く知られている。我々は、免疫系の完成した成熟マウスへの接種によって重篤な免疫不全症を伴う急性の白血病を起こすフレンドレトロウイルス複合体を用い、gag遺伝子産物に対する免疫応答の誘導によりウイルス感染細胞の排除を誘導することが可能であることを示した。本年度はフレンドレトロウイルスgag遺伝子産物上で細胞障害性T細胞(CTL)が認識する抗原構造を同定するため、gag遺伝子の部分断片を組換えワクシニアウイルスを用いて発現させ、ウイルス感染細胞に特異的なCTLクローンがGag蛋白質のどの部分を認識するかを解析した。その結果、調べた全てのCTLクローンはgag遺伝子5'端に近い、細胞表面型Gag蛋白のリーダー配列部分に認識構造を持つことがわかった。更に、この部分のアミノ酸配列に基づく合成ペプチドを多数作成し、それらを吸着した標的細胞の障害反応を解析した結果、多くのCys残基を含む疎水性のペプチドが認識エピトープとして同定された。このエピトープは、zinc finger motifの一部分に極めて類似した構造を持ち、現在までに知られている他のCTLエピトープとは全く異なる構造を示していた。現在、このCTL認識エピトープを含む合成ペプチドワクチンを用いた感染防御実験が進行中である。
  • 文部科学省：科学研究費補助金(奨励研究(A))
    研究期間 : 1993年 -1993年 
    代表者 : 宮沢 正顯; 宮澤 正顕
     
    血栓性血小板減少性紫斑病(TTP)は、血小板減少・溶血性貧血・腎障害を伴う原因不明の症候群で、現在まで再現性のある動物モデルがなく、その発症メカニズムは不明のままであった。我々は、ヒトの自己免疫症のモデル動物として広く用いられているMRL/lprマウスから新たに樹立した、内在性レトロウイルスenv遺伝子産物に特異的なモノクローナル抗体の中に、正常マウスへの移入によって血小板減少と全身の出血斑形成を起こす致死性のクローン36D1.1を見いだした。このクローンのハイブリドーマ細胞を同系正常マウスに移入後経時的に静脈血を採血し、末梢血中に各血球数を調べた。その結果、細胞移入後3日目から5日目にかけて、血小板数のみが移入前に比較して有意に減少して行き、赤血球数・白血球数には有意な変化は見られなかった。一方、移入マウスの脾臓にはヘモジデリンの過剰沈着を認め、更に電子顕微鏡的に赤脾髄や肺の血管内に血小板の異常凝集像を認めた。これらin vivoの解析結果は、上記ハイブリドーマ移入マウスの体内で血小板が異常な凝集反応を起こして消費されていることを示唆した。そこで、正常マウス血漿から分離した血小板を用い、in vitroで上記ハイブリドーマの培養上清と反応させたところ、対照の抗体には見られない凝集反応を認めた。更に、分離血小板を用いた蛍光抗体法でも、上記抗体が血小板に結合することが示唆された。この病原性ハイブリドーマの培養上清中に分泌された抗体分子は、IgGクラスに属するにも関わらずプロテインAに対する結合能が非常に弱かった。そこで、イオン交換法を用いて抗体分子を精製し、現在正常マウスへの静注移入による病変形成の有無を検討中である。
  • 文部科学省：科学研究費補助金(重点領域研究)
    研究期間 : 1992年 -1992年 
    代表者 : 宮沢 正顯; 宮澤 正顕
     
    マウスに重篤な免疫不全症を伴え急性の赤白血病を誘発するフレンド白血病レトロウイルス複合体に対し、レトロウイルスenv或いはgag遺伝子を発現する組換えワクシニアウイルスで感染防御免疫が誘導できることを昨年度迄に証明した。その後、gag遺伝子の部分発現により、感染防御免疫の誘導に必要な抗原エピトープは、N-末のp15蛋白質上に存在することが明らかとなった。そこで、p15蛋白質上でヘルパーT細胞認識エピトープとなり得る両親和性構造を探したところ、ヒト免疫不全ウイルスGag蛋白質の矢張りN-末に位置するp17と相同性のある構造を持ったペプチドが最も両親和性が高いことがわかった。そこで、この両親和性部分を含む合成ペプチド(Len-Ser-Leu-Thr-Leu-Asp-His-Trp--Lys-Arg-Trp-Cys)を作り、アジュバントとともにマウスを免疫して、フレンドウイルスに対する感染防御免疫の誘導の有無を調べた。実験にはフレンドウイルスに高感受性の(B10.AxA)F_1マウスと、中等度に感受性の(B10.AxA.BY)F_1又は(B10xA)F_1マウスを用いた。免疫した(B10.AxA)F_1マウスの約半数で、フレンドウイルス接種後60日目迄に、一旦発症した白血病性脾腫の退縮が見られた。対照のアジュバントのみで免疫したマウスは全例が発症を免れた。約半数の個体で感染防御が成立したという結果は、組換えワクシニアウイルスでの成績と一致する。現代、ペプチドの投与量や経路を種々に変えて実験を続行中である。
  • レトロウイルス感染に対する防御免疫反応の宿主遺伝子による制御
    科学研究費補助金
    研究期間 : 1992年
  • レトロウイルス感染に対する宿主免疫応答の抗原特異性と認識エピトープ構造
    科学研究費補助金
    研究期間 : 1992年
  • 自己免疫病モデルマウス由来抗gp70モノクローナル抗体による腎炎・血管炎の形成
    科学研究費補助金
    研究期間 : 1992年
  • Host genetic control of protective immune responses against retroviral infections
    Health and Labour Sciences Research Grants
    研究期間 : 1992年
  • Antigen specificity and epitope structures in host immune responses to retroviruses
    Grants and Funding
    研究期間 : 1992年
  • Pathogenicity of antiretroviral gp70 monoclonal antibodies established from autoimmune strains of mice
    Grants and Funding
    研究期間 : 1992年
  • 文部科学省：科学研究費補助金(重点領域研究)
    研究期間 : 1991年 -1991年 
    代表者 : 宮沢 正顯; 宮澤 正顕
     
    ヒトレトロウイルス感染症に対する有効な予防ワクチンの開発を最終目標に、その最大の問題点であるウイルス被膜抗原の急速な変異に対処するため、より変異の少ないコア抗原による感染防御免疫の誘導を試みた。今年度はマウスに重篤な免疫不全症を伴う急性白血病を誘発するフレンド白血病レトロウイルスをモデルとして用い、分子クロ-ン化したコア抗原遺伝子及びその部分断片を発現する組換えワクシニアウイルスを使って動物を免疫することにより、感染防御免疫が誘導されるか否かを調べた。その結果、以下の諸点を明らかにする成果を得た:1.フレンド白血病ヘルパ-レトロウイルスのコア抗原をコ-ドするgag遺伝子は、少なくともふたつの翻訳開始部位を同一の読み枠上に持ち、そのうち上流側のCTGコドンを含むようにこの遺伝子を発現ベクタ-に組換えた場合のみ、感染細胞表面にコア抗原が発現する。2.フレンド白血病ヘルパ-ウイルスコア抗原を発現する組換えワクシニアウイルスを用いてマウスを免疫することにより、CD4陽性のヘルパ-T細胞が抗原特異的に感作される。3.上記によってコア抗原特異的免疫能を獲得した動物は、フレンドウイルス複合体を接種すると一旦発症した白血病性脾腫が急速に退縮する。この際、コア抗原特異的に免疫された動物ではウイルス感染後IgGクラスのウイルス中和抗体の産生を認めるが、無関係な抗原で免疫された対照群のマウスはIgGクラスの中和抗体は産生しない。4.gag遺伝子部分断片の発現実験から、感染防御に有効な抗原エピト-プはコア蛋白質前駆体N末のp15蛋白上に存在することが明らかとなった。以上の結果を踏まえ、今後合成ペプチドを用いてp15蛋白上の抗原エピト-プを同定する実験を続けて行く予定である。
  • 自己免疫病の病因解明
  • Molecular identification of host genetic factors determining HIV-exposed but uninfected status and its application for AIDS prevention and therapy
  • Analysis on the pathogenesis of autoimmune diseases
  • Vaccine development aiming at HIV infection

委員歴

  • 2011年04月 - 現在   科学技術振興機構   専門委員
  • 1990年 - 現在   日本病理学会   学術評議員   日本病理学会
  • 2006年04月 - 2023年03月   日本学術振興会   専門委員
  • 2005年 - 2012年   大学基準協会   相互評価委員

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