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.

本研究旨在建立临床肺和心肺联合移植（LTX和HLTX）急性排斥反应监测系统。为此，检查了来自受体的淋巴细胞的细胞毒性。从支气管肺泡灌洗液（BAL）中分离淋巴细胞，并将其用作标准细胞介导的淋巴细胞溶解（CML）试验中的效应细胞，该试验使用一系列B淋巴母细胞系（LCL）作为靶标。平均溶解百分比（i. e.当LCL靶标共有一种或多种HLA I类致敏供体同种异体抗原（“I类相关靶标”）时，与无排斥或CMV感染相比，在无巨细胞病毒（CMV）感染的急性排斥期间，LCL靶标的细胞毒性显著更高。特别是在病理工作公式分类为“A1”的急性排斥反应的早期阶段，其显著更高。在CMV感染无排斥反应期间，LCL靶细胞的平均溶解百分比无显著差异。本研究为LTX和HLTX患者急性排斥反应的临床和组织学诊断提供了一种功能性的体外试验方法。然后，检测肺实质细胞的特性，以探讨LTX急性排斥反应的机制。在体外建立了永生化的人气道上皮细胞系和同种异体淋巴细胞的混合培养来模拟LTX。在混合淋巴细胞-气道上皮细胞系（ML-1）中，AECL），细胞毒性淋巴细胞（CTL）没有被激发。结果发现，AECL刺激淋巴细胞产生白细胞介素（IL）-10，IL-2刺激ML-AECL产生CTL活性。该CTL对AECL具有MHC Ⅰ类特异性细胞毒作用，提示肺实质细胞是炎性细胞因子环境下抗原提呈细胞的候选细胞。

项目成果

Kawauchi M, Nakajima J, 他3名: "Contribution of T cells in concordant xenoheart rejection." Transplantation Proceedings. 26. 1193-1194 (1994)

Kawauchi M、Nakajima J 和其他 3 人：“T 细胞在异种心脏排斥反应中的贡献。” 26. 1193-1194 (1994)

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）

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)。

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。​​”

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。