Data_Sheet_1_Functional Change of Effector Tumor-Infiltrating CCR5+CD38+HLA-DR+CD8+ T Cells in Glioma Microenvironment.docx

Human glioma facilitates an impaired anti-tumor immunity response, including defects in circulation of T lymphocytes. The level of CD8+ T-cell activation acts as an immune regulator associated with disease progression. However, little is known about the characteristics of peripheral and tumor-infiltrating CD8+ T cells in patients with glioma. In this study, we examined the level of CD8+ T-cell activation in a group of 143 patients with glioma and determined that peripheral CD3+ T cells decreased in accordance with disease severity. The patients' peripheral CD8+ T-cell populations were similar to that of healthy donors, and a small amount of CD8+ tumor-infiltrating lymphocytes was identified in glioma tissues. An increase in activated CD8+ T cells, characterized as CD38+HLA-DR+, and their association with disease progression were identified in the patients' peripheral blood and glioma, and shown to display enriched CCR5+ and TNFR2+ expression levels. Ex vivo examination of CD38+HLA-DR+CD8+ T cells indicated that this subset of cells displayed stronger secretion of IFN-γ and IL-2 before and after a 6-h stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin (ION) relative to healthy CD38+HLA-DR+CD8+ T cells, indicating the functional feasibility of CD38+HLA-DR+CD8+ T cells. Higher CCL5 protein and mRNA levels were identified in glioma tissues, which was consistent with the immunohistochemistry results revealing both CCL5 and CD38+HLA-DR+CD8+ T cell expression. Patients' CCR5+CD38+HLA-DR+CD8+ T cells were further validated and shown to display increases in CD45RA+CCR7− and T-bet+ accompanied by substantial CD107-a, IFN-γ, and Granzyme B levels in response to glioma cells.

人脑胶质瘤加剧了抗肿瘤免疫反应的障碍，包括T淋巴细胞循环的缺陷。CD8+ T细胞激活水平作为一种与疾病进展相关的免疫调节因子。然而，关于胶质瘤患者外周和肿瘤浸润性CD8+ T细胞的特征，了解甚少。在本研究中，我们检测了143名胶质瘤患者的CD8+ T细胞激活水平，并发现随着疾病严重程度的增加，外周CD3+ T细胞数量减少。患者的周围CD8+ T细胞群体与健康供体相似，并在胶质瘤组织中发现了少量CD8+ 肿瘤浸润性淋巴细胞。在患者的周围血液和胶质瘤中，发现了激活的CD8+ T细胞（特征为CD38+HLA-DR+）的增加，以及它们与疾病进展的相关性，并显示出富含CCR5+和TNFR2+的表达水平。体外检测CD38+HLA-DR+CD8+ T细胞表明，与健康的CD38+HLA-DR+CD8+ T细胞相比，该细胞亚群在用佛波酯-12-肉豆蔻酸-13-乙酸（PMA）和离子霉素（ION）进行6小时刺激前后表现出更强的IFN-γ和IL-2分泌，这表明CD38+HLA-DR+CD8+ T细胞的功能可行性。胶质瘤组织中CCL5蛋白和mRNA水平较高，与免疫组化结果一致，揭示了CCL5和CD38+HLA-DR+CD8+ T细胞的表达。患者的CCR5+CD38+HLA-DR+CD8+ T细胞进一步得到验证，并显示在胶质瘤细胞的刺激下，CD45RA+CCR7-和T-bet+的增加，伴随着显著的CD107-a、IFN-γ和Granzyme B水平升高。