Stervbo Ulrik, Seidel Maximilian, Uszkoreit Julian, Kaliszczyk Sviatlana, Anft Moritz, Eisenacher Martin, Westhoff Timm H, Babel Nina
Center for Translational Medicine and Immune Diagnostics Laboratory, Medical Department I, Marien Hospital Herne, University Hospital of the Ruhr-University Bochum, Herne, Germany.
Berlin Institute of Health, Berlin-Brandenburg Center for Regenerative Therapies, and Institute of Medical Immunology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany.
Front Transplant. 2025 Apr 24;4:1537656. doi: 10.3389/frtra.2025.1537656. eCollection 2025.
Kidney transplantation is associated with an increased risk of severe COVID-19 disease. Additionally, cells of the kidney express ACE-2 making them a potential target of the SARS-CoV-2 virus. Both uncontrolled viral replication and T-cell receptor (TCR) mediated cellular immunity towards the infected cells could lead to tissue destruction in the kidney. In cases where pathological findings are not always capable of providing definitive diagnosis, insights into the TCR repertoire could offer valuable information. Here we present a case of potentially infection driven tubulitis in a kidney transplant patient.
The source of kidney infiltrating T-cells was assessed through next generation TCR sequencing. Using cells from the living donor and overlapping peptide pool of SARS-CoV-2 S-, N-, and M-protein (Wuhan variant), antigen specific T-cells were isolated from peripheral blood by overnight stimulation and subsequent isolation using antibodies and magnetic beads against CD154 and CD137. The clonotypes of these two samples were compared to the clonotypes in a single kidney biopsy cylinder.
We found that 11.1% of the repertoire of the kidney infiltrating T cells were identical to SARS-CoV-2 specific T-cells in the periphery, and only 3.1% of the repertoire was identical to allo-specific TCRs. We also observed substantial overlap between the TCR repertoires of virus-specific and donor-specific T cells, with high similarity and even identical TCR sequences present in both populations. The TCRs with dual specificity constituted a larger proportion of the allo-specific than the virus specific population. These results indicate that SARS-CoV-2 specific T-cells may directly spill into an allo-specific T cell response and that either class of T-cells may cause the observed tubulitis.
TCR-seq of whole biopsies is a method to evaluate the ingragraft TCR repertoire can complement routine pathology and provide further insights into the mechanisms underlying a diagnosis.
肾移植与严重 COVID-19 疾病风险增加相关。此外，肾细胞表达血管紧张素转换酶 2（ACE-2），使其成为严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）病毒的潜在靶点。不受控制的病毒复制和 T 细胞受体（TCR）介导的针对受感染细胞的细胞免疫都可能导致肾脏组织破坏。在病理结果并不总是能够提供明确诊断的情况下，了解 TCR 库可能会提供有价值的信息。在此，我们报告一例肾移植患者中可能由感染驱动的肾小管炎病例。
通过下一代 TCR 测序评估肾脏浸润性 T 细胞的来源。利用来自活体供体的细胞以及 SARS-CoV-2 S 蛋白、N 蛋白和 M 蛋白（武汉变种）的重叠肽库，通过过夜刺激从外周血中分离出抗原特异性 T 细胞，随后使用针对 CD154 和 CD137 的抗体及磁珠进行分离。将这两个样本的克隆型与单个肾活检样本中的克隆型进行比较。
我们发现，肾脏浸润性 T 细胞库中有 11.1%与外周血中 SARS-CoV-2 特异性 T 细胞相同，仅有 3.1%与同种异体特异性 TCR 相同。我们还观察到病毒特异性和供体特异性 T 细胞的 TCR 库之间存在大量重叠，两个群体中存在高度相似甚至相同的 TCR 序列。具有双重特异性的 TCR 在同种异体特异性群体中所占比例大于病毒特异性群体。这些结果表明，SARS-CoV-2 特异性 T 细胞可能直接混入同种异体特异性 T 细胞反应中，并且这两类 T 细胞中的任何一类都可能导致观察到的肾小管炎。
全活检的 TCR 测序是一种评估移植物内 TCR 库的方法，可补充常规病理学检查，并为诊断背后的机制提供进一步的见解。
