Data Sheet 1_Immune responses in rodent whole eye transplantation: elucidation and preliminary investigations into rejection diagnosis and monitoring.pdf

BackgroundWhole Eye Transplantation (WET) offers potential for vision restoration but is hindered by the complex challenge of immune rejection. Understanding and closely monitoring these immunological responses is crucial for advancing WET. This study delves into the timeline and nature of immune responses in a rodent model of WET without immunosuppression, aiming to elucidate a detailed picture of the immune landscape post-transplantation and establish innovative diagnostic and monitoring methods.
MethodsWe employed a multi-faceted approach to analyze immune responses post-WET, including assessments of gross changes in corneal transparency, thickness, and skin condition. Histopathological examinations of both ocular and surrounding skin tissues provided insights into cellular changes, complemented by ocular RT-qPCR for molecular analysis. Serological analysis was employed to quantify cytokines, chemokines, and donor-specific antibodies, aiming to identify potential biomarkers correlating with WET rejection and to validate the presence of antibody-mediated rejection. These methodologies collectively contribute to the development of non-invasive diagnostic and monitoring strategies for WET.
ResultsOur study revealed a rapid and acute immune response following WET, characterized by an early innate immune response dominated by complement involvement, and infiltration of neutrophils and monocytes by post-operative day (POD) 2. This was succeeded by an acute T-cell-mediated immune reaction, predominantly involving T helper 1 (Th1) cells and cytotoxic T lymphocytes (CTLs). The presence of donor specific antibody (DSA) and indications of pyroptosis in the early phases of rejection were observed. Notably, the early elevation of serum CXCL10 by POD4, coupled with ocular CD3+ cell infiltration, emerged as a potential early biomarker for WET rejection. Additionally, corneal transparency grading proved effective as a non-invasive monitoring tool.
ConclusionThis study offers a first-time comprehensive exploration of immune responses in WET, unveiling rapid and complex rejection mechanisms. The identification of early biomarkers and the development of non-invasive monitoring techniques significantly advance our understanding of WET rejection. Additionally, these findings establish an essential baseline for future research in this evolving field.

研究背景：全眼移植（Whole Eye Transplantation, WET）可为视力恢复提供潜在可能，但受限于免疫排斥这一复杂难题。深入理解并密切监测这类免疫应答，对于推进全眼移植研究至关重要。本研究针对无免疫抑制条件下的全眼移植啮齿类动物模型，探究其免疫应答的时序与特征，旨在阐明移植后免疫微环境的详细全貌，并建立创新性的诊断与监测方法。
研究方法：我们采用多维度分析手段，对全眼移植后的免疫应答展开研究，具体包括角膜透明度、厚度及皮肤状态的大体形态评估。对眼部及周围皮肤组织进行组织病理学检查，以解析细胞层面的病理变化，并辅以眼部实时荧光定量聚合酶链反应（RT-qPCR）开展分子水平分析。通过血清学分析定量检测细胞因子、趋化因子及供体特异性抗体，旨在筛选与全眼移植排斥反应相关的潜在生物标志物，并验证抗体介导排斥反应的存在。上述多种实验方法共同助力全眼移植无创诊断与监测策略的开发。
研究结果：本研究揭示了全眼移植后快速且剧烈的免疫应答进程：早期以补体激活为主的固有免疫应答，并于术后第2天（post-operative day, POD）出现中性粒细胞与单核细胞浸润；随后以T辅助1（T helper 1, Th1）细胞和细胞毒性T淋巴细胞（Cytotoxic T Lymphocytes, CTLs）为核心的急性T细胞介导免疫反应接踵而至。研究同时观察到供体特异性抗体（Donor Specific Antibody, DSA）的存在，以及排斥反应早期阶段的细胞焦亡迹象。值得注意的是，术后第4天血清CXCL10的早期升高，联合眼部CD3+细胞浸润，可作为全眼移植排斥反应的潜在早期生物标志物。此外，角膜透明度分级作为无创监测工具被证实具备良好应用价值。
研究结论：本研究首次对全眼移植中的免疫应答展开全面系统的探索，揭示了快速且复杂的排斥反应机制。早期生物标志物的筛选与无创监测技术的开发，极大提升了我们对全眼移植排斥反应的认知水平。此外，本研究成果为该新兴领域的后续研究奠定了重要基础。