Development of the new monitoring system for acute rejection in clinicallung and lung-heart transplantation, focusing on the cytotoxicity specific to the major histocompatibility complex class I antigens of the donor organ.

开发用于临床肺和肺心移植中急性排斥反应的新监测系统，重点关注供体器官主要组织相容性复合物 I 类抗原的特异性细胞毒性。

• 批准号: 06557070
• 负责人: NAKAJIMA Jun
• 金额: $ 4.54万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557070/
• 关键词: lung transplantation; acute rejection; bronchoalveolar lavage fluid; cytotoxic T cell; major histocompatibility complex; B-lymphoblastoid cell line; interleukin-10; mixed lymphocyte culture; 凍結保存; 気道上皮細胞; BEAS-2B; flow cytometry; programmed fr

This research aimed to establish the monitoring system for acute rejection in cliniacal lung and heart-lung transplantation (LTX and HLTX). For this purpose, cytotoxicity of the lymphocytes derived from the recipients were examined. Lymphocytes were isolated from bronchoalveolar lavage (BAL) and used as effector cells in standard cell-mediated lymphocytolysis (CML) assays using a series of B-lymphoblastoid cell lines (LCL) as targets. Mean percent lysis (i. e. Cytotoxicity) of LCL targets was significantly higher during acute rejection without cytomegaloviral (CMV) infection compared to no rejection or CMV infection when these targets shared one or more of the HLA class I sensitizing donor alloantigens ("class I-relevant targets"). Especially it is significantly higher in the early phase of acute rejection classified as "A1" by the pathological working formulation. There was no significant difference in mean percent lysis of LCL targets during CMV infection without rejection. This study provides a functional in vitro assay to further support the clinical and histological diagnosis of acute rejection in the LTX and HLTX patients.The properties of lung parenchymal cells were then examined to investigate the mechanisms of acute rejection in LTX.In vitro mixed culture of an immortal human airway epithelial cell line and allogeneic lymphocytes was made up to simulate the LTX.In mixed lymphocyte-airway epithelial cell line (ML-AECL), cytotoxic lymphocytes (CTL) were not elicited. We found that interleukin (IL)-10 was secreted from the lymphocytes with the stimulation of the AECL.CTL activity was generated when the ML-AECL was cultured with IL-2. This CTL showed the MHC-class I-specific cytotoxicity against the AECL.It is therefore suggested that lung parenchymal cells were candidates for antigen-presenting cells in the circumstance of inflammatory cytokines.

这项研究旨在建立临床肺和心肺移植（LTX和HLTX）急性排斥反应的监测系统。为此，检查了从接受者得出的淋巴细胞的细胞毒性。从支气管肺泡灌洗（BAL）中分离出淋巴细胞，并用一系列B林型细胞细胞系（LCL）作为靶标，用作标准细胞介导的淋巴细胞解析（CML）测定法中的效应细胞。与没有排斥反应或CMV感染相比，急性排斥反应期间，LCL靶标的平均裂解百分比（e。细胞毒性）明显更高。尤其是在病理工作制定的急性排斥早期阶段，急性排斥反应的早期较高。 CMV感染期间LCL靶标的平均裂解百分比没有显着差异。这项研究提供了一种功能性的体外测定，以进一步支持LTX和HLTX患者急性排斥反应的临床和组织学诊断。然后检查肺实质细胞的特性，以研究LTX中急性排斥的机制。在不成熟的人类气管上皮细胞线的体外混合培养物。未引起淋巴细胞 - 气管上皮细胞系（ML-AECL），细胞毒性淋巴细胞（CTL）。我们发现，当用IL-2培养ML-AECL时，生成了淋巴细胞中的白介素（IL）-10，并从淋巴细胞中分泌。该CTL显示针对AECL的MHC级I特异性细胞毒性。因此，在炎症细胞因子的情况下，肺实质细胞是抗原呈递细胞的候选者。

项目成果

吉竹 毅、中島 淳,他: "日本猿における一側同種肺移植後慢性移行期の移植肺および脾臓における主要組適合性抗原(MHC)発現の検討" 日本呼吸器外科学会雑誌. 8. 548-554 (1994)

Takeshi Yoshitake、Jun Nakajima 等人：“日本猴单侧同种异体肺移植后慢性过渡期移植肺和脾脏中主要相容性抗原 (MHC) 表达的检查”日本呼吸外科学会杂志 8。 548-554（1994）

Nakajima J,Ono M,et al.: "Human allogeneic airway epithelial cell line induced lymphocyte to secrete interleukin-10" Transplantation Proceedings. 28(3). 1861-1862 (1996)

Nakajima J,Ono M,et al.：“人同种异体气道上皮细胞系诱导淋巴细胞分泌白细胞介素 10”移植论文集。

中島 淳,他: "結節性硬化症に合併した過誤腫性肺脈管筋腫症-特異な臨床修飾が加えられた1例-" 日本胸部疾患学会雑誌. 33. 80-84 (1995)

Jun Nakajima 等人：“错构瘤性肺血管肌瘤病并发结节性硬化症 - 具有独特临床修改的病例”日本胸科疾病学会杂志 33. 80-84 (1995)。

Kawauchi M,Nakajima J,et al.: "Which is the target vessel of lung rejection?A primate heart-lung transplantation study" Transpl.antation Proceedings. 26(4). 2338-2339 (1994)

Kawauchi M，Nakajima J，et al.：“哪一个是肺排斥的靶血管？灵长类心肺移植研究”移植论文集。

Nakajima J,Ono M,et al.: "The role of costimulatory molecules on an airway epithelial cells acting as an alloantigenpresenting cells." Transplantation Proceedings. in press. (1997)

Nakajima J、Ono M 等人：“共刺激分子对作为同种抗原呈递细胞的气道上皮细胞的作用。​​”