Kawashima Mitsuaki, Oliver Jillian D, Watanabe Tatsuaki, Oishi Hisashi, Huang Ning, Konoeda Chihiro, Hirayama Shin, Hwang David M, Li Qixuan, Huszti Ella, Liu Mingyao, Keshavjee Shaf, Juvet Stephen, Martinu Tereza
Latner Thoracic Research Laboratories, Toronto Lung Transplant Program, Ajmera Transplant Centre, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario, Canada.
Department of Laboratory Medicine and Molecular Diagnostics, Sunnybrook Hospital, Toronto, Ontario, Canada.
JHLT Open. 2023 Dec 29;4:100050. doi: 10.1016/j.jhlto.2023.100050. eCollection 2024 May.
The mouse orthotopic single lung transplant (LTx) model is an important scientific tool to explore LTx immunology. C57BL/10J (B10, H-2b) to C57BL/6J (B6, H-2b) minor alloantigen-mismatched LTx exhibits mild acute rejection and chronic fibrosis, mimicking human LTx, where acute rejection is dampened by immunosuppressants and chronic lung allograft dysfunction (CLAD) develops over time. However, we have observed variations in allograft histology across experiments, which were not explained by animal vendor or experimental conditions. The purpose of this study was to evaluate those variations objectively.
We performed a retrospective review of B10-to-B6 LTx performed in our laboratory 2012-2019. Only LTx without experimental interventions (eg, immunomodulatory agents or genetic modifications) examined at day 28 was eligible for this study. Mice from each surgeon were selected and divided into 3 groups to represent early, middle, and late timepoints in their mouse LTx experience (143 LTx from 5 surgeons). Histology from these LTx was graded in a randomized and blinded manner. Pathological variations and trajectories were graphed; logistic regression analyses were performed for statistical assessment.
Distribution and trajectories of pathological outcomes were significantly different across surgeons. In multivariable logistic regression analyses, surgeon was associated with pathological outcomes whereas case number was not. Longer warm ischemia time was associated with more severe pleural fibrosis.
The B10 to B6 single LTx model can be a powerful tool to recapitulate CLAD-like histology. However, this is a challenging operation and surgeon-dependent variability in histopathological findings needs to be taken into account when designing experimental protocols.
小鼠原位单肺移植（LTx）模型是探索肺移植免疫学的重要科学工具。C57BL/10J（B10，H-2b）到C57BL/6J（B6，H-2b）的次要同种异体抗原不匹配的肺移植表现出轻度急性排斥反应和慢性纤维化，类似于人类肺移植，其中急性排斥反应可通过免疫抑制剂得到缓解，而慢性肺移植功能障碍（CLAD）会随着时间的推移而发展。然而，我们观察到不同实验中同种异体移植组织学存在差异，这些差异无法用动物供应商或实验条件来解释。本研究的目的是客观评估这些差异。
我们对2012年至2019年在我们实验室进行的B10到B6肺移植进行了回顾性研究。只有在第28天检查时没有实验干预（如免疫调节剂或基因修饰）的肺移植符合本研究要求。从每位外科医生的小鼠中进行选择并分为3组，以代表他们在小鼠肺移植经验中的早期、中期和晚期时间点（来自5位外科医生的143例肺移植）。对这些肺移植的组织学进行随机和盲法分级。绘制病理变化和轨迹图；进行逻辑回归分析以进行统计学评估。
不同外科医生的病理结果分布和轨迹存在显著差异。在多变量逻辑回归分析中，外科医生与病理结果相关，而病例数则无关。较长的热缺血时间与更严重的胸膜纤维化相关。
B10到B6单肺移植模型可以成为重现CLAD样组织学的有力工具。然而，这是一项具有挑战性的手术，在设计实验方案时需要考虑组织病理学结果中外科医生依赖的变异性。
