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Abstract：
Isolation of tumor-specific T cells and their antigen receptors (TCRs) from malignant pleural effusions (MPE) may facilitate the development of TCR-transduced adoptive cellular immunotherapy products for advanced lung cancer patients. However, the characteristics and markers of tumor-specific T-cells in MPE are largely undefined. To this end, to establish the phenotypes and antigen specificities of CD8+ T cells, we performed single-cell RNA and TCR sequencing of samples from three advanced lung cancer patients. Dimensionality reduction on a total of 4,983 CD8+ T cells revealed 10 clusters including naïve, memory, and exhausted phenotypes. We focused particularly on exhausted T cell clusters and tested their TCR reactivity against neoantigens predicted from autologous cancer cell lines. Four different TCRs specific for the same neoantigen and one orphan TCR specific for the autologous cell line were identified from one of the patients. Differential gene expression analysis in tumor-specific T cells relative to the other T cells identified CXCL13, as a candidate gene expressed by tumor-specific T cells. In addition to expressing CXCL13, tumor-specific T cells were present in a higher proportion of T cells co-expressing PDCD1(PD-1)/TNFRSF9(4-1BB). Furthermore, flow cytometric analyses in advanced lung cancer patients with MPE documented that those with high PD-1/4-1BB expression have a better prognosis in the subset of 57 adenocarcinoma patients (p = .039). These data suggest that PD-1/4-1BB co-expression might identify tumor-specific CD8+ T cells in MPE, which are associated with patients\' prognosis. (233 words).
摘要：
从恶性胸腔积液（MPE）中分离肿瘤特异性T细胞及其抗原受体（TCR）可能有助于开发用于晚期肺癌患者的TCR转导过继性细胞免疫治疗产品。然而,MPE中肿瘤特异性T细胞的特征和标志物在很大程度上是不明确的.为此,建立CD8+T细胞的表型和抗原特异性，我们对3例晚期肺癌患者的样本进行了单细胞RNA和TCR测序.总共4,983个CD8+T细胞的维度减少显示10个簇，包括幼稚，记忆,和耗尽的表型。我们特别关注耗尽的T细胞簇，并测试了它们对自体癌细胞系预测的新抗原的TCR反应性。从患者之一中鉴定出对相同新抗原具有特异性的四种不同TCR和对自体细胞系具有特异性的一种孤儿TCR。肿瘤特异性T细胞相对于其他T细胞的差异基因表达分析将CXCL13鉴定为由肿瘤特异性T细胞表达的候选基因。除了表达CXCL13之外,肿瘤特异性T细胞存在于较高比例的共表达PDCD1(PD-1)/TNFRSF9(4-1BB)的T细胞中。此外，对MPE晚期肺癌患者的流式细胞仪分析表明,PD-1/4-1BB高表达者在57例腺癌患者亚组中预后较好(p=0.039).这些数据表明PD-1/4-1BB共表达可能在MPE中鉴定肿瘤特异性CD8+T细胞，与患者预后相关。（233字）
