Blocking antibodies targeting immune checkpoint molecules achieved invaluable success in tumor therapy and amazing clinical responses in a variety of cancers. Although common treatment protocols have improved overall survival in patients with chronic lymphocytic leukemia (CLL), they continue to relapse and progress. In the present in vitro study, the application of anti-PD-1 and anti-TIM-3 blocking antibodies was studied to restore the function of exhausted CD8⁺ T cells in CLL. CD8⁺ T cells were isolated from peripheral blood of 20 patients with CLL, treated with blocking antibodies, and cocultured with mitomycin-frozen non-CD8⁺ T cell fraction as target cells. Cultures were stimulated with anti-CD3/CD28 antibodies to assess the proliferation of CD8⁺ T cells by MTT and stimulated with PMA/ionomycin to measure the levels of CD107a expression and cytokine production by flow cytometry and ELISA, respectively. Our results showed that the blockade of PD-1 and TIM-3 does not improve the proliferation of CD8⁺ T cells in CLL patients. No significant difference was found between control and blocked groups in terms of degranulation properties and production of IFN-γ, TNF-α, IL-2, and IL-10 by CD8⁺ T cells. We observed that pre-treatment of CD8⁺ T cells with blocking antibodies in CLL patients at early clinical stages had no effects on restoring their functional properties. Further in vitro and in vivo complementary studies are required to more explore the utility of checkpoint inhibitors for CLL patients.

针对免疫检查点分子的阻断抗体在肿瘤治疗中取得了宝贵的成功，并在各种癌症中取得了惊人的临床反应。尽管常见的治疗方案提高了慢性淋巴细胞白血病 (CLL) 患者的总体生存率，但它们会继续复发和进展。在目前的体外研究中，研究了抗PD-1和抗TIM-3阻断抗体的应用，以恢复CLL中衰竭的CD8 ⁺ T细胞的功能。从 20 名 CLL 患者的外周血中分离出CD8 ⁺ T 细胞，用封闭抗体处理，并与丝裂霉素冷冻的非 CD8 ⁺ T 细胞部分作为靶细胞共培养。用抗 CD3/CD28 抗体刺激培养物以评估 CD8 的增殖⁺通过 MTT 检测并用 PMA/离子霉素刺激的 T 细胞，分别通过流式细胞术和 ELISA 测量 CD107a 表达水平和细胞因子产生水平。我们的结果表明，阻断 PD-1 和 TIM-3 并不能改善CLL 患者中 CD8 ⁺ T 细胞的增殖。在脱颗粒特性和 CD8 ⁺ T 细胞产生 IFN-γ、TNF-α、IL-2 和 IL-10 方面，对照组和阻断组之间没有发现显着差异。我们观察到 CD8 ^{+ 的}预处理在早期临床阶段，CLL 患者体内带有阻断抗体的 T 细胞对恢复其功能特性没有影响。需要进一步的体外和体内补充研究，以更多地探索检查点抑制剂对 CLL 患者的效用。