The cuff leak test (CLT) has been widely accepted as a simple and noninvasive method for predicting post-extubation stridor (PES). However, its accuracy and clinical impact remain uncertain. We aimed to evaluate the reliability of CLT and to assess the impact of pre-extubation variables on the incidence of PES. A prospective observational study was performed on adult critically ill patients who required mechanical ventilation for more than 24 h. Patients were extubated after the successful spontaneous breathing trial, and CLT was conducted before extubation. Of the 191 patients studied, 26 (13.6%) were deemed positive through CLT. PES developed in 19 patients (9.9%) and resulted in a higher reintubation rate (8.1% vs. 52.6%, p < 0.001) and longer intensive care unit stay (8 [4.5-14] vs. 12 [8-30.5] days, p = 0.01) than patients without PES. The incidence of PES and post-extubation outcomes were similar in patients with both positive and negative CLT results. Compared with patients without PES, patients with PES had longer durations of endotracheal intubation and required endotracheal suctioning more frequently during the 24-h period prior to extubation. After adjusting for confounding factors, frequent endotracheal suctioning more than 15 times per day was associated with an adjusted odds ratio of 2.97 (95% confidence interval, 1.01-8.77) for PES. In conclusion, frequent endotracheal suctioning before extubation was a significant PES predictor in critically ill patients. Further investigations of its impact on the incidence of PES and patient outcomes are warranted.

BACKGROUND: Benzodiazepines are widely used for anesthesia and sedation and have immunomodulatory properties that may negatively influence clinical outcomes; however, the cellular targets and intermediary signaling pathways involved are unclear. We examined the immunomodulatory effects of the benzodiazepine midazolam on human macrophages and associated molecular mechanisms. METHODS: We analyzed effects of midazolam pretreatment on lipopolysaccharide (LPS)-induced upregulation of the costimulatory molecule CD80 and secretion of the pro-inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α, interleukin-10, and nitric oxide (NO) in the human monocyte-macrophage cell line THP-1 and in peripheral monocyte-derived macrophages (PMDMs). The effects of midazolam on NF-κB, IκBα protein, and mitogen-activated protein kinase (MAPK) activation were analyzed in THP-1 cells. We analyzed the involvement of translocator protein (TSPO) in the immunomodulatory effects of midazolam using TSPO ligands. The role of TSPO was investigated using THP-1 cells overexpressing TSPO and THP-1 cells with TSPO knockdown through transfection with small interfering RNA for TSPO. RESULTS: Midazolam suppressed LPS-induced upregulation of CD80 and release of IL-6 and NO in THP-1 cells and PMDMs. Additionally, midazolam suppressed the activation of NF-κB/AP-1 and MAPKs in human THP-1 cells. The assessed synthetic TSPO ligands showed the same inhibitory effects on macrophage activation as midazolam. Macrophages overexpressing TSPO exhibited enhanced susceptibility to immunosuppression by midazolam, and macrophages lacking TSPO expression exhibited reduced effects of midazolam. CONCLUSION: Midazolam inhibits LPS-stimulated immune responses in human macrophages by activating TSPO signaling. Suppression of macrophage activity may contribute to deleterious side effects of benzodiazepines reported in critically ill patients.

Objective: The authors assessed the efficacy and safety of landiolol, an ultra-short-acting beta-blocker, with those of amiodarone in the restoration of sinus rhythm for postoperative atrial fibrillation (POAF) in intensive care unit (ICU) patients. Design: A retrospective data analysis. Setting: Data were collected from patients admitted to the ICU in a single university hospital between 2012 and 2015. Participants: Records of a total of 276 patients who developed POAF after ICU admission were collected from hospital records. Interventions: None. Measurements and Main Results: Treatment success was defined as restoration of sinus rhythm without concomitant therapy within 24 hours of treatment and lasting for more than an hour. The landiolol dosage was in the range of 0.7 mu g/kg/min-to-2.5 mu g/kg/min. The authors compared a total of 55 patients with POAF who received either landiolol (n = 32) or intravenous amiodarone (n = 23) in the ICU. The major findings were that the median time required for conversion to sinus rhythm was shorter in landiolol patients compared with amiodarone patients (75 v 150 min respectively, p = 0.0355). However, treatment success rates did not differ significantly after 24 hours (odds ratio 1.25, 95% confidence interval 0.17-9.09, p = 0.60). Adverse events with bradycardia leading to drug discontinuation were seen only in the patients receiving amiodarone (n = 3, p = 0.032). Conclusions: Landiolol achieved swift and safe restoration of sinus rhythm in ICU patients with POAF and could be considered as a favorable drug choice over amiodarone in such patients. (C) 2016 Elsevier Inc. All rights reserved.

Background: Haloperidol, a tranquilizing agent, is administered both to treat symptoms of psychotic disorders and to sedate agitated and delirious patients. Notably, haloperidol has been suggested to inhibit the immune response through unknown mechanisms. We hypothesized that the sedative modulates the immune response via NF-κB. Material/Methods: Using flow cytometry, we analyzed the effects of haloperidol on expression CD80 and CD86 in RAW 264 cells and in primary macrophages derived from bone marrow. Secretion of interleukin (IL)-1β, IL-6, and IL-12 p40 was measured by enzyme-linked immunosorbent assay. In addition, NF-κB activation was evaluated using a reporter assay based on secretory embryonic alkaline phosphatase. Finally, synthetic antagonists were used to identify the dopamine receptor that mediates the effects of haloperidol. Results: Haloperidol inhibited NF-κB activation, and thereby suppressed expression of CD80, as well as secretion of IL-1b, IL-6, and IL-12 p40. CD80 and IL-6 levels were similarly attenuated by a D2-like receptor antagonist, but not by a D1-like receptor antagonist. Conclusions: The data strongly suggest that haloperidol inhibits the immune response by suppressing NF-κB signaling via the dopamine D2 receptor.

Background: Haloperidol, a tranquilizing agent, is administered both to treat symptoms of psychotic disorders and to sedate agitated and delirious patients. Notably, haloperidol has been suggested to inhibit the immune response through unknown mechanisms. We hypothesized that the sedative modulates the immune response via NF-kappa B. Material/Methods: Using flow cytometry, we analyzed the effects of haloperidol on expression CD80 and CD86 in RAW 264 cells and in primary macrophages derived from bone marrow. Secretion of interleukin (IL)-1 beta, IL-6, and IL-12 p40 was measured by enzyme-linked immunosorbent assay. In addition, NF-kappa B activation was evaluated using a reporter assay based on secretory embryonic alkaline phosphatase. Finally, synthetic antagonists were used to identify the dopamine receptor that mediates the effects of haloperidol. Results: Haloperidol inhibited NF-kappa B activation, and thereby suppressed expression of CD80, as well as secretion of IL-1 beta, IL-6, and IL-12 p40. CD80 and IL-6 levels were similarly attenuated by a D2-like receptor antagonist, but not by a D1-like receptor antagonist. Conclusions: The data strongly suggest that haloperidol inhibits the immune response by suppressing NF-kappa B signaling via the dopamine D2 receptor.

BACKGROUND: Haloperidol has immunomodulatory effects when used to treat patients with. schizophrenia and also is used to sedate critically ill patients in the intensive care unit. Although the mechanism by which haloperidol affects immune function is unclear, one possibility is that it alters dendritic cell (DC) function. DCs are potent antigen-presenting cells that influence the activation and maturation of T lymphocytes. In this study, we investigated the in vitro and in vivo immunomodulatory effects of haloperidol on DC-mediated immune responses. METHODS: Using bone marrow derived DCs in cell culture, we evaluated the effect of haloperidol on expression of costimulatory molecules (CD80 and CD86), major histocompatibility complex class II molecules, and the DC maturation marker CD83. DC culture supernatants also were evaluated for interleukin-12 p40 levels. In addition, we analyzed the effect of haloperidol on a mixed cell culture containing DCs and lymphocytes and measured the secretion of interferon-gamma in the culture supernatants. We also assessed the in vivo effects of haloperidol on hapten-induced contact hypersensitivity responses. RESULTS: Haloperidol inhibited the expression of CD80, CD86, major histocompatibility complex class II, and CD83 molecules on DCs and the secretion of interleukin-12p40 in DC culture supernatants. In mixed cell cultures containing both T cells (CD4(+) and CD8 alpha(+)) and DCs, haloperidol-treated DCs suppressed the proliferation of allogeneic T cells and effectively inhibited the production of interferon-gamma. In vivo, haloperidol reduced hapten-induced contact hypersensitivity responses. Furthermore, an antagonist to D2-like receptor suppressed the maturation of DCs in a manner similar to haloperidol. CONCLUSIONS: The results of our study suggest that haloperidol suppresses the functional maturation of DCs and plays an important role in the inhibition of DC-induced T helper 1 immune responses in the whole animal. Furthermore, the effect of haloperidol on DCs may be mediated by dopamine D2-like receptors. Together, these results demonstrate that administration of haloperidol suppresses DC-mediated immune responses.

Background: Although Acute Kidney Injury Network (AKIN) staging is widely used, it has been suggested that classification using serum creatinine levels, which fluctuate because of fluid balance, is not always appropriate for acute kidney injury (AKI) detection. We hypothesized that some patients are misdiagnosed as having no AKI due to dilution resulting from intraoperative infusion, and have worse outcomes than typical patients with no AKI.Methods: We retrospectively selected patients who did not fulfill the AKI criteria from those who underwent cardiac surgery and remained in an intensive care unit (ICU) for ≥7 days. The patients were divided into two groups: those with AKI (AKI group) and those without AKI (no-AKI group), classified using serum creatinine levels adjusted for fluid balance during the perioperative period. We compared the characteristics and outcomes of the two groups.Results: After adjustment for serum creatinine, 7 of 26 patients were categorized as having AKI. The AKI group had significantly fewer ventilator-free days during a 28-day period and significantly longer ICU stays than the no-AKI group (5.86 ± 10.0 days vs. 15.6 ± 9.71 days, respectively, P = 0.050 36.4 ± 20.6 days vs. 14.9 ± 10.7 days, respectively, P = 0.033).Conclusion: Adjustment of creatinine level for perioperative fluid balance could improve the accuracy of AKI diagnosis after cardiac surgery.

OBJECTIVE: To evaluate optimal humidifier water temperature when using a helmet for noninvasive ventilation. METHODS: Twenty-eight healthy individuals underwent 8 cm H2O CPAP ventilation with FIO2 of 0.21 and 0.5. Each was sequentially tested in the following order: Using the helmet without humidification at ambient temperature with humidification with unheated chamber water and with humidification with the chamber water at 31°C, 34°C, and 37°C. At each setting, after a 20 min stabilization period, measurements were taken. Comfort level at each setting was evaluated using a visual analog scale rated zero (least comfortable) to 10 (most comfortable). RESULTS: Temperature and relative and absolute humidity inside the helmet increased however, the comfort scores significantly decreased as the humidification chamber water temperature increased. Regardless of the FIO2, statistically significantly highest comfort scores were obtained when humidification water, with and without active humidification, was at ambient temperature. Unacceptable absolute humidity was obtained only without humidification at room temperature when FIO2 was 0.5. CONCLUSIONS: With the clinical use of a helmet, for patient comfort and mucosal humidification during CPAP, the most desirable conditions are likely to be obtained by humidifying without heating, that is by leaving the water in the humidifier chamber at room temperature. © 2013 Daedalus Enterprises.

Background: Dendritic cells (DCs), as antigen-presenting cells, play a key role in the induction and regulation of adaptive immune response. Midazolam is reported to have immunomodulatory properties that affect immune cells. However, the effect of midazolam on DCs has not been characterized. We examined the immunomodulatory properties of midazolam on DC-mediated immune response. Methods: After allowing murine bone marrow-derived DCs induced by granulocyte macrophage colony stimulating factor to mature, we analyzed their expression of costimulatory molecules (CD80 and CD86), major histocompatibility complex class II molecules, and the secretion of interleukin-12 p40. In vitro, we evaluated the effect of midazolam on maturing DCs in mixed cell cultures containing DCs and T cells. In vivo, we investigated the contact-hypersensitivity response. Results: Midazolam suppressed the expression of CD80, CD86, and major histocompatibility complex class II molecules from murine DCs. Treated with midazolam, DCs also secreted less interleukin-12 p40. In mixed cell cultures with CD3-positive T cells, midazolam-treated DCs showed less propensity to stimulate the proliferation of CD3-positive T cells and the secretion of interferon-gamma from CD4-positive T cells. Midazolam-treated DCs impaired the induction of contact-hypersensitivity response. Treatment with ligands for peripheral benzodiazepine receptor inhibited the up-regulation of CD80 during DC maturation. Conclusion: Midazolam inhibits the functional maturation of murine DCs and interferes with DC induction of T helper 1 immunity in the whole mouse. In addition, it appears that the immunomodulatory effect of midazolam is mediated via the action of midazolam on the peripheral benzodiazepine receptor.

This is a report of the case of an 18 year-old woman who was admitted to a psychiatric hospital for anorexia nervosa. After admission, hypoglycemic coma developed as a complication, and although she recovered from the coma in response to intravenous glucose administration, an ultrasonic echocardiogram (UCG) demonstrated takotsubo cardiomyopathy. Because the patient's serum phosphate level had decreased to 0.9 mg·dl⁻¹, she was diagnosed with refeeding syndrome and admitted to the ICU. Artificial feeding (both enteral and parenteral), early supplementation with phosphorous, and other electrolytes were started at a reduced calorific rate. The patient recovered, and she was discharged to a general ward on ICU day 16. When nutrition is started after prolonged starvation, it is important to start at a reduced calorific rate and to monitor vital signs, electrolyte levels, and water balance closely in order to prevent refeeding syndrome.

BACKGROUND: Dendritic cells (DCs) play a key role as antigen-presenting cells and growing evidence Suggests that DCs influence T-cell activation and regulate the polarity of the immune response. Ketamine has been reported to have immunomodulatory properties that affect immune cells, including macrophages and natural killer cells. However, the effect of ketamine on DCs has not been characterized. We examined the immunomodulation of DCs by ketamine. METHODS: We used bone marrow-derived DCs induced by granulocyte-monocytecolony stimulating factor and interleukin (IL)-4 from bone marrow and analyzed the expression of costimulatory molecules (CD40, CD80, and CD86), major histocompatibility complex class II molecules, and secretion of IL-12p40. Furthermore, we evaluated the immune response in mixed cell cultures of DCs and T cells and the contact hypersensitivity response in a whole animal. RESULTS: Ketamin suppressed the expression of CD40, CD80, and major histocompatibility complex class 11 molecules in DCs. DCs treated with ketamine also secreted less IL-12p40 and displayed greater endocytosis. In mixed cell cultures with CD4(+) T cells and DCs, ketamine-treated DCs showed less propensity to stimulate the proliferation of CD4(+) T cells and the secretion of interferon from CD4(+) T cells. Furthermore, ketamine-treated DCs impaired the induction Of a cell-mediated immune response. CONCLUSION: Our findings suggest that ketamine inhibits the functional maturation of DCs and interferes with DC induction of Th1 immunity in the whole animal. These novel findings provide new insight into the immunopharmacological role of ketamine. (Anesth Analg 2009:109:793-800)

Respiratory complication is common after a repair of thoracic aneurysm, although tracheal compression caused by hematoma and felt strips following surgery is a rare cause. We report the case of a patient who experienced difficult weaning from ventilator after a repair of a thoracic aortic aneurysm and was diagnosed as a tracheal compression outside of trachea revealed by bronchoscopy and chest CT scan. Re-operation was successfully performed to relieve the compression under monitoring by bronchoscopy. Patient was disconnected from the ventilator three weeks after the reoperation and transferred to a rehabilitation hospital.

Transfusion-related acute lung injury (TRALI) is a serious complication of transfusion of blood and blood components. TRALI is reported to be the most common cause of transfusion-associated death. TRALI has increasingly become known in the medical community. However, TRALI has been overlooked frequently because of poor knowledge of medical staffs. We report two cases of TRALI after massive transfusion due to massive bleeding during cardiovascular surgery. In the perioperative period, the diagnosis of TRALI is difficult to make because of coexistence of various factors leading to hypoxia. Thus, in this report, we discuss the management of these two cases focusing upon the differential diagnosis of TRALI.

A 31-year-old woman suffering from bronchiolitis obliterans received bilateral living-donor lung transplantation to treat end-stage respiratory failure. After 5 days' mechanical ventilation, the patient was successfully extubated. During mechanical ventilation, the patient was sedated with a continuous intravenous infusion of propofol and dexmedetomidine (DEX). To assuage postoperative pain, morphine was infused, first intravenously, then epidurally. The administration of DEX was continued after extubation to prevent agitation. After the administration of epidural morphine was discontinued on day 10 in the intensive care unit (ICU), the patient complained of pain in the oral cavity. Greater pain was reported after the discontinuation of DEX, and symptoms of tachycardia and dyspnea appeared. A dermatologist diagnosed the oral symptoms as herpetic stomatitis, and a course of treatment with aciclovir was begun. A continuous infusion of DEX was again started on the same day, and was continued until ICU day 13. During the administration of DEX, the oral cavity pain was bearable. The patient was successfully discharged from the ICU on ICU day 13. We conclude that DEX could be used to provide analgesia for herpetic stomatitis after living-donor lung transplantation, at a dosage that achieves appropriate sedation.

We describe advanced hemodynamic insufficiency and remarkably high myoglobinemia in a 77-year-old man who was admitted to the intensive care unit after total aortic arch replacement. Serum myoglobin showed an unusually high value (peak value, 155 030 ng.ml(-1)). The patient died of sepsis and untreatable metabolic acidosis. Pseudomonas aeruginosa was detected in blood culture specimens after his death. On histopathological examination, dense congregations of gram-negative bacilli were present in clots in blood vessels, while congregations of gram-negative bacilli around the circumference of small blood vessels were particularly apparent in every specimen examined. Moreover, a generalized breakdown of muscle fibers, consistent with findings of rhabdomyolysis, was observed in muscle tissue throughout the body.

Forbidden CD4+ββ T cells, which produce interleukin (IL)-4 predominantly, are a pathological subset in the development of colitis in T-cell receptor α chain (TCRα)-deficient mice. Stimulation of naive CD4+ T cells with IL-4 induces Th2 development via the activation of signal transducers and activators of transcription (STAT) 6. In the present study, we had found that IL-4 enhanced the expression of STAT6 in CD4+ββ T cells isolated from TCRα-/- mice with colitis, suggesting that the IL-4 signal in the CD4+ κκ T cells is mediated by STAT6. To further investigate the role of STAT6 in the development of colitis induced by TCRα deficiency, we generated double-deficient mice by crossing TCRα-/- mice and STAT6-/- mice. Surprisingly, STAT6 deficiency did not result in decreased severity of colitis in TCRα-/- mice. STAT6-deficient CD4+ ββ T cells produced IL-4 and intraperitoneal injection of anti-IL-4 monoclonal antibody in the nondiseased TCRα-/- and STAT6 double-deficient mice prevented the colitis formation, thus indicating that the cells differentiated into the Th2 phenotype have the ability to mediate the development of the colitis in the absence of STAT6.

Intestinal intraepithelial lymphocytes (IELs), which reside between the basolateral faces of intestinal epithelial cells (IECs), provide a first-line defense against pathogens via their cytotoxic activity. Although IEC-derived IL-7 and IL-15 are key regulatory cytokines for the development and activation of IELs, we report here that IL-15 but not IL-7 mediates the reciprocal interaction between IELs and IECs, an important interaction for the regulation of appropriate mucosal immunohomeostasis. IL-15-treated IELs induced cell death in IECs via the cytotoxic activity in vitro. Among the different subsets of IL-15-treated IELs, CD4(-)CD8(-)TCR(-) lELs, which express NK marker (DX5 or NK1.1), showed the most potent syngenic IEC killing activity. These intraepithelial NK cells expressed Ly-49 molecules, NKG2 receptors, and perforin. These results suggest the possibility that the cell death program of IECs could be regulated by self-produced IL-15 through the activation of intraepithelial NK cells.

OBJECTIVE: To investigate, in a rat model, the role of the Mac-1/ICAM-1 pathway and the anti-inflammatory activity of steroid in ventilator-induced lung injury. DESIGN: Prospective, randomized controlled study. SETTING: Animal investigation using Wistar rats. INTERVENTION: Rats in three randomly assigned groups of 18, a total of 54 animals, were subject to the following: Two groups received high peak inspiratory pressure (35 cm H2O) ventilation after pretreatment with methylprednisolone (high-methylprednisolone group) or pretreatment with methylprednisolone vehicle (high-vehicle group). The third group of animals received low peak inspiratory pressure (7 cm H2O) ventilation after pretreatment with methylprednisolone vehicle (low-vehicle group). Except for animals previously killed to establish baseline values, after 40 mins of mechanical ventilation, the animals in each group were killed. Some animals provided histological samples, and the rest received total lung lavage. MEASUREMENT: We measured flow cytometry of lavage fluid, cell counts of tissue samples, and pressure-volume curves before and after mechanical ventilation. RESULTS: In the groups that received high peak inspiratory pressure ventilation, both the number of neutrophils that infiltrated the lungs and the expression of Mac-1 and ICAM-1 on neutrophils and macrophages increased significantly more than in the low-vehicle group. Static lung compliance was reduced in the high peak inspiratory pressure groups. In the high peak inspiratory pressure groups, there were significantly fewer neutrophils in samples from the high-methylprednisolone group (0.412 +/- 0.1 x 10(5)) than from the high-vehicle group (1.10 +/- 0.1 x 10(5); p < .05). The high-vehicle group showed greater expression of CD11b on neutrophils, but this was significantly decreased by methylprednisolone (mean fluorescence intensity: high-vehicle, 118.4 +/- 34.3; high-methylprednisolone, 25.8 +/- 4.2; p < .05). The lung mechanics measured by pressure-volume curve analysis were deteriorated less in the high-methylprednisolone group. CONCLUSION: Our study suggests that a neutrophil-endothelium interaction via the Mac-1/ICAM-1 pathway is involved in the activation and recruitment of neutrophils in ventilator-induced lung injury. Activation and recruitment of neutrophils were lessened by pretreatment with methylprednisolone, which might have contributed to the improvement of lung dysfunction after mechanical ventilation.

Activated neutrophils contribute to the development of ventilator-induced lung injury (VILI) caused by high-pressure mechanical ventilation. However, exact cellular and molecular mechanisms have not been conclusively studied. Our investigation aimed to examine expression of adhesion molecules by both neutrophils and macrophages in lung lavage fluids of rats with VILI. Further, involvement of proinflammatory (tumor necrosis factor-alpha) and profibrogenetic (transforming growth factor-beta 1) mediators was analyzed at mRNA level in lung tissue. Wistar rats were ventilated by high pressure (45 cm H(2)O of peak inspiratory pressure, n = 23) or low pressure (7 cm H(2)O, n = 13) with 0 positive end-expiratory pressure. After 40 min of comparative ventilation, lung lavage was performed in 20 rats from the experimental group and 10 from the control for immunofluorescence analysis with anti-Mac-1 and anti-ICAM-1 monoclonal antibodies. The lung tissues from remaining rats were subjected to pathological and reverse transcription-polymerase chain reaction examinations. Although there was no significant change of PaO(2) in the low-pressure group, PaO(2) was decreased in the high-pressure group. The high-pressure group also had greater neutrophil infiltration into alveolar spaces, upregulation of CD54 and CD11b on alveolar macrophages, and more transforming growth factor-beta 1 mRNA in lung tissues. Tumor necrosis factor-alpha was not involved in the pathogenesis of the severe VILI observed. Histologic findings also demonstrated more infiltrating neutrophils, destructive change of the alveolar wall, and deposition of matrix in the high-pressure group. These results suggest that a series of proinflammatory reactions and profibrogenetic process may be involved in the course of VILI.

本年度は骨髄由来の樹状細胞に対するハロペリドールの作用と免疫応答に与える影響を検討した。GM-CSFの存在下にマウスの骨髄細胞 より誘導した樹状細胞をハロペリドールで処理すると、通常の樹状細胞が呈する分化成熟の過程が抑制された。この過程には副刺激分子CD80, CD86の発現とTh1を誘導するサイトカインであるIL-12の産生が含まれる。これらをハロペリドールの処理は抑制した。次にこのようにハロペリドールによって形質変化した樹状細胞がT細胞のプライミングに影響をあたえるのかをリンパ球混合培養法によって解析を行った。このアッセイによって試験管内てハロペリドールで処理した樹状細胞はリンパ球のプライミングを抑制することが示された。次にハロペリドールの作用が動物個体レベルのの免疫応答に影響しうるのかを解析した。この目的で樹状細胞の免疫によって 誘導できる接触型過敏症モデルによる解析を導入した。この疾患モデルは典型的なTh1型免疫応答によって起こる疾患モデルでありこ のモデルを用いた解析を行うことで動物個体レベルのTh1型免疫応答への影響を解析することが可能となる。結果はハロペリドールで処理した樹状細胞によって免疫を行うと対照群と比較して接触過敏症が抑制され、Th1型免疫応答が抑制されることが解明された

We examined the immunomodulatory effects of the benzodiazepine midazolam on human macrophages and associated molecular mechanisms. We analyzed effects of midazolam pretreatment on LPS-induced upregulation of the costimulatory molecule and the pro-inflammatory factors in THP-1 and in PMDMs. The effects of midazolam on NF-κB, and MAPK activation were analyzed in THP-1 cells. The role of TSPO was investigated using THP-1 cells overexpressing TSPO and with TSPO knockdown through transfection with small interfering RNA for TSPO. Results Midazolam suppressed upregulation of CD80 and release of IL-6 and NO in THP-1 cells and PMDMs. Midazolam suppressed the activation of NF-κB/AP-1 and MAPKs in human THP-1 cells. Macrophages overexpressing TSPO exhibited enhanced susceptibility to immunosuppression by midazolam, and macrophages lacking TSPO expression exhibited reduced effects of midazolam. Midazolam inhibits LPS-stimulated immune responses in human macrophages by activating TSPO signaling.

We analyzed the effects of haloperidol on dendritic cell (DC) and DC-mediated immune response. haloperidol suppressed the maturation of immature DCs induced by LPS. Furthermore, we analyzed the effect of haloperidol on DC-mediated immune response by the use of contact hypersensitivity model of mouse. Haloperidol-treated DC suppressed the development of contact hypersensitivity in mouse. This model is assumed to be developed by typical Th1 immune response. Out results strongly suggest that haloperidol suppress the maturation of immature DCs and the development of Th1 type immune response. In addition, the inhibitory effect was mediated by dopamine D2-like receptor via the inhibition of NF-κB in immune cells such as macrophages and DCs.

Benzodiazepine shows sedative effect that suppresses central nerve system via GABA receptor. On the other hand benzodiazepine has another functional molecule as translocator protein (TSPO). Benzodiazepine was reported that affects function of immune cells, but it is not uncertain about those of functional mechanism. We report that we found the mechanism of midazolam on immune cells. Using a murine macrophage cell line, RAW264 cells, we analyzed the effect of midazolam on the expression of CD80 and CD86 and the secretion of IL-6. Midazolam suppressed the expression of costimulatory molecules and IL-6. The activation of RAW264 was inhibited by TSPO ligand but not by GABA legends. Finally we analyzed the contribution of TSPO by analysis of using knockdown RAW264 cells by siRNA for TSPO. The effect of midazolam was inhibited by knockdown of TSPO. These results showed that midazolam has an inhibitory action of immune response on murine macrophages via TSPO.

Dendritic cells (DCs), as antigen-presenting cells, play a key role in the induction and regulation of adaptive immune response. Propofol is reported to have immunomodulatory properties that affect immune cells. However, the effect of propofol on DCs has not been characterized. We examined the immunomodulatory properties of propofol on DC-mediated immune response. Propofol promoted the functional maturation of murine DCs and augmented DC induction of T helper 1 immunity in the whole mouse. In addition, it appears that the immunostimulatory effect of propofol is attributed to the component of lipid emulsion rather than anesthetic propofol itself.

Midazolam is a widely-used sedative drug in critical care and anesthesia. We analyzed the effect of midazolam on the function of dendritic cells. Midazolam suppressed the functional maturation of dendritic cells by lipopolysaccharidein terms of the expression of costimulatory molecules and IL-12. Midazolam-treated dendritic cells inhibited the allostimulatory activity of dendritic cells. Midazolam-treated dendritic cells have reduced activity inducing contact hypersensitivity response, one of typical cell-mediated immune responses. It was suggested that these inhibitory effects of midazolam is mediated by the effect on peripheral benzodiazepine receptor (PBR) by the study using PBR-specific ligand.

We analyzed the immunosuppressive state after recovery from sepsis induced by peritonitis in mice. We mainly used experimental system analyzing antigen-specific immune-response induced by mucosal immunization method. In mice recovered from sepsis, antigen-specific immune-response was depressed in terms of cellular immunity. Furthermore, we tried the improvement of depressed immune-response by intervention into PD-1 and PD-L1 signalling.

Midazolam inhibits the functional maturation of murine DCs and interferes with DC induction of T helper 1 immunity in the whole mouse. Further, by the use of rabbit model of acute lung injurty, we elucidated the role of maintaining spontaneous breathing for the progression of acute lung injury. Even in the lung-protective strategy for acute lung injury, the inadequate maintenance of spontaneous ventilation can make clinical outcome worse.

Prior to the assessment of anti-cytokine therapy, we tried to elucidate the effect of pCO2 on the production of proinflammatory cytokine in order to determine the effect of pCO2 in HFO upon experimental results in animal study. Wistar rats were ventilated and administered HCl intratracheally. After HC1 injection, rats were randomized to two groups: hypercapnia group and normocapnia group. The animals were ventilated with HFO for 4hrs. Lung volume was titrated to achieve PaCO2 40-50 mmHg (normocapnia group) or 80mmHg (hypercapnia group). After 4hrs of mechanical ventilation, the level of TNF-a,IL-6,and CINC-1 in BAL fluid was determined. No significant difference was observed between these two groups. However, W/D ratio and histological study showed that hypercapnia attenuated lung injury. Furthermore, we tried to establish quantitative RT-PCR system of chemokine receptors(CCR4,5,6,7,9,CXCR3,4) by the use of colitis model. Chemokine receptors except CCR6 were upregulated in diseased mice. Interestingly, mRNA of CCR7 and CXCR4 were upregulated in diseased colon compared with non-diseased small intestine.

A 敗血症下の粘膜免疫応答を調査する目的で、特定の細菌の毒素が消化管における炎症に与える影響を検討した。昨年度は消化管におけるTh1型炎症に対する影響を調査したが、本年度は消化管におけるTh2型の炎症反応をどのように修飾するかを調査した。この目的でマウスにおける経口抗原に対するアレルギーモデルを用いた検討を行った。Pertussis toxin(百日咳毒素)(以下PTと略)の投与が、アレルギーモデルの発症ににどのように影響するかを調べた。このアレルギーモデルの最も重要な症状は抗原を投与して2時間以内に起こる激しい水様性下痢である。PT投与を行なうと下痢の発症を100%阻害できることを見出した。今後はこのアレルギーの病態がどう修飾されているかを血中抗原特異的IgEの値、消化管粘膜に存在する好酸球の動態、脾臓と消化管粘膜におけるリンパ球の産生動態から更なる解析を行う。 B 樹状組胞に対する麻酔、鎮静薬の影響を調査した。本年度は骨髄からGM-CSF, IL-4の存在下に分化させた樹状細胞に対するケタミンの影響を調査した。骨髄由来の樹状細胞をケタミンの存在下で培養すると、樹状細胞上に発現される1型の主要組織適応抗原と更に副刺激分子であるCD40の発現が著しく抑制されることを示した。更にケタミン存在下で培養した樹状細胞がallo抗原に対する反応をどのように変えるかを調査して本年度の来年度の日本麻酔科学会で報告する予定である C 更に集中治療臨床で、粘膜免疫系と関わる合併症の実状を調べるために、大阪府立急性期総合医療センターとの共同研究により、昨年度に構築した大動脈弓手術後患者のデータベースを用いて、術後合併症の予知因子や術後経過への影響を統計学的に探索し、今まで不明であった大動脈瘤術後の合併症の予知因子、術後経過への影響を始めて明らかにした(Journal of Vascular Surgery 2006,Journal of Vascular Surgery in press)。