Adaptation of Imaging Mass Cytometry to Explore the Single Cell Alloimmune Landscape of Liver Transplant Rejection.
Adaptation of Imaging Mass Cytometry to Explore the Single Cell Alloimmune Landscape of Liver Transplant Rejection.
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DOI：
10.3389/fimmu.2022.831103
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发表时间：
2022
影响因子：
7.3
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中科院分区：
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排斥反应仍然是实体器官移植中移植物丢失的一个重要原因，但由于临床活检标本的稀缺，对移植物内同种免疫的深入探索受到限制。新兴的单细胞免疫分析技术在识别自身免疫和癌症免疫生物学机制方面显示出了希望。在这些应用中，成像质量细胞计数（IMC）已被证明能够在固定的组织标本内进行高度多重的单细胞免疫表型分析。在这项研究中，开发了一个由10个经验证的标志物组成的IMC组，以探索IMC在表征从肝移植受者获得的临床组织样本中慢性排斥（CR）的免疫景观方面的可行性。IMC染色具有高度特异性，与传统免疫组织化学相当。开发了一个单细胞分割分析管道，可以对109，245个离散细胞（包括30，646个免疫细胞）进行详细的可视化和定量。CR标本中的特异性降低鉴定出11个独特的免疫亚群。在CR中，大多数免疫亚群增加并且空间相关，包括两个CD 45 +/CD 3 +/CD 8+细胞毒性T细胞群和离散的CD 68+巨噬细胞群，这在无排斥反应（NR）的肝脏中未观察到。通过主成分分析和逻辑回归的建模显示，单细胞数据可用于构建具有高度一致性的统计模型（Wilcoxon秩和检验，p=0.000036）。这项研究强调了IMC在临床排斥事件中以单细胞分辨率研究同种免疫微环境的能力。IMC的进一步验证有可能检测新的生物标志物，识别治疗靶点，并生成实体器官移植临床结局的患者特异性预测模型。
Rejection continues to be an important cause of graft loss in solid organ transplantation, but deep exploration of intragraft alloimmunity has been limited by the scarcity of clinical biopsy specimens. Emerging single cell immunoprofiling technologies have shown promise in discerning mechanisms of autoimmunity and cancer immunobiology. Within these applications, Imaging Mass Cytometry (IMC) has been shown to enable highly multiplexed, single cell analysis of immune phenotypes within fixed tissue specimens. In this study, an IMC panel of 10 validated markers was developed to explore the feasibility of IMC in characterizing the immune landscape of chronic rejection (CR) in clinical tissue samples obtained from liver transplant recipients. IMC staining was highly specific and comparable to traditional immunohistochemistry. A single cell segmentation analysis pipeline was developed that enabled detailed visualization and quantification of 109,245 discrete cells, including 30,646 immune cells. Dimensionality reduction identified 11 unique immune subpopulations in CR specimens. Most immune subpopulations were increased and spatially related in CR, including two populations of CD45+/CD3+/CD8+ cytotoxic T-cells and a discrete CD68+ macrophage population, which were not observed in liver with no rejection (NR). Modeling via principal component analysis and logistic regression revealed that single cell data can be utilized to construct statistical models with high consistency (Wilcoxon Rank Sum test, p=0.000036). This study highlights the power of IMC to investigate the alloimmune microenvironment at a single cell resolution during clinical rejection episodes. Further validation of IMC has the potential to detect new biomarkers, identify therapeutic targets, and generate patient-specific predictive models of clinical outcomes in solid organ transplantation.