阻断CD40-CD40L信号通路对小鼠肺移植术后闭塞性细支气管炎的影响

doi:10.3969/j.issn.1006-7795.2018.02.019

摘要/Abstract

摘要： 目的本研究使用抗CD40L抗体（克隆号：MR1）阻断CD40-CD40L信号通路，利用小鼠原位左肺移植模型，对比其与传统免疫抑制剂环孢素/甲泼尼龙对小鼠术后移植物闭塞性细支气管炎（obliterative bronchiolitis，OB）的治疗效果，并进一步评估其对移植物术后损伤的影响及相应机制，为临床肺移植术后治疗方法提供新思路。方法构建小鼠原位左肺移植模型，以野生型Balb/c小鼠作为供体小鼠，野生型C57BL/6小鼠作为受体小鼠。移植术后10 d，牺牲小鼠，采用苏木精/伊红（hematoxylin-eosin，HE）染色、Masson染色以及中性粒细胞特异性染色（抗髓过氧化物酶），观察各组移植左肺的组织病理学改变，利用实时荧光定量聚合酶链式反应（polymerase chain reaction，PCR）技术检测移植物中白细胞介素-17A（interleukin-17A，IL-17A） mRNA的表达水平的变化。结果术后10 d，注射了环孢素/甲泼尼龙和抗CD40L抗体治疗的小鼠，移植左肺外观体积增大；病理结果显示抗CD40L抗体治疗组较环孢素/甲泼尼龙治疗组炎性细胞浸润和OB现象得到有效缓解（P<0.05），中性粒细胞浸润显著减少（P<0.01）；抗CD40L抗体治疗组移植物中IL-17A mRNA表达水平，较环孢素/甲泼尼龙治疗组显著降低（P<0.01）。结论利用抗CD40L抗体阻断CD40-CD40L信号通路能够有效保护移植物细支气管上皮，缓解小鼠肺移植术后急性排斥和OB反应以及小气道损伤，且效果优于传统抑制剂环孢素/甲泼尼龙。

关键词: 小鼠原位左肺移植, 抗CD40L抗体(MR1), 环孢素/甲泼尼龙, 闭塞性细支气管炎

Abstract: Objective Our research focus on the effects to obliterative bronchiolitis (OB) by blocking CD40-CD40L pathway with anti-mCD40L on allografts in mouse orthotopic lung transplantation model, and comparing with traditional immunosuppressive agents-cyclosporine/methylprednisolone (C/M) to explore new clinical treatment methods. Methods Murine orthotopic lung transplantation was performed in wild type C57BL/6 mice using Balb/c donors. At day 10, histopathologic characteristics were assessed by hematoxylin-eosin (HE) staining, Masson staining and neutrophil staining with antimyeloperoxidase; real-time polymerase chain reaction (RT-PCR) was performed for interleukin-17A (IL-17A) expressions. Results Donor lung of recipients were harvested at 10 days after transplantation. Groups with treatment of anti-mCD40L and cyclosporine/methylprednisolone showed an enlargement appearance compared with allograft group. Pathology results demonstrated that administration of anti-mCD40L can better improve rejection injury and OB compared with cyclosporine/methylprednisolone treatment group (P<0.05). Furthermore, administration of anti-mCD40L significantly reduced transcription levels of IL-17A in grafts(P<0.01). Conclusion This findings demonstrated that by blocking CD40-CD40L pathway with anti-mCD40L can better alleviate transplant rejection and OB mediated by IL-17A in mouse lung transplantation.

Key words: mouse left orthotopic lung transplantation, anti-mCD40L, cyclosporine/methylprednisolone, obliterative bronchiolitis(OB)

中图分类号:

R392.4

陈荣卷, 陈其瑞, 许江南, 丁跃中. 阻断CD40-CD40L信号通路对小鼠肺移植术后闭塞性细支气管炎的影响[J]. 首都医科大学学报, 2018, 39(2): 258-264.

Chen Rongjuan, Chen Qirui, Xu Jiangnan, Ding Yuezhong. Effects to obliterative bronchiolitis by blocking CD40-CD40L pathway in the mouse lung transplantation[J]. Journal of Capital Medical University, 2018, 39(2): 258-264.

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