State Key Laboratory of Genetic Engineering, And Key Laboratory of Medical Molecular Virology of Ministry of Education/Health, School of Life Sciences, Fudan University, Shanghai, 200438, China.
Shanghai Institute of Infectious Disease and Biosecurity, School of Life Sciences, Fudan University, Shanghai, 200438, China.
Virol Sin. 2023 Apr;38(2):285-295. doi: 10.1016/j.virs.2023.01.003. Epub 2023 Jan 16.
HIV-specific chimeric antigen receptor (CAR) T-cells have been developed to target HIV-1 infected CD4 T-cells that express HIV Env proteins. However, T cell exhaustion and the patient-specific autologous paradigm of CAR-T cell hurdled clinical applications. Here, we created HIV-specific CAR-T cells using human peripheral blood mononuclear cells and a 3BNC117-E27 (3BE) CAR construct that enabled the expression of programmed cell death protein (PD-1) -blocking scFv E27 and the single-chain variable fragment of the HIV-1-specific broadly neutralizing antibody 3BNC117 to target native HIV Env. Compared with T cells expressing 3BNC117-CAR alone, 3BE CAR-T cells showed greater cytotoxic activity against HIV Env cells with stronger proliferation capability, higher killing efficiency, and enhanced cytokine secretion in the presence of HIV Env-expressing cells. Furthermore, we manufactured TCR-deficient 3BE CAR-T cells through gene editing and demonstrated that these CAR-T cells could effectively kill HIV Env cells in vivo without the occurrence of severe graft-versus-host disease (GvHD) in NSG mice. These data suggest that we have provided a feasible approach to the generation of "off-the-shelf" anti-HIV CAR-T cells in combination with PD-1 checkpoint blockade immunotherapy, which can be a powerful therapeutic candidate for the functional cure of HIV.
HIV 特异性嵌合抗原受体（CAR）T 细胞已被开发用于靶向表达 HIV Env 蛋白的 HIV-1 感染的 CD4 T 细胞。然而，T 细胞耗竭和 CAR-T 细胞的患者特异性自体范式阻碍了临床应用。在这里，我们使用人外周血单核细胞和 3BNC117-E27（3BE）CAR 构建体创建了 HIV 特异性 CAR-T 细胞，该构建体能表达程序性细胞死亡蛋白（PD-1）阻断 scFv E27 和 HIV-1 特异性广泛中和抗体 3BNC117 的单链可变片段，以靶向天然 HIV Env。与单独表达 3BNC117-CAR 的 T 细胞相比，3BE CAR-T 细胞对表达 HIV Env 的细胞具有更强的细胞毒性活性、更强的增殖能力、更高的杀伤效率和增强的细胞因子分泌。此外，我们通过基因编辑制造了 TCR 缺陷型 3BE CAR-T 细胞，并证明这些 CAR-T 细胞在没有发生严重移植物抗宿主病（GvHD）的情况下可以有效地杀死体内的 HIV Env 细胞。这些数据表明，我们已经提供了一种可行的方法来生成与 PD-1 检查点阻断免疫疗法相结合的“现成”抗 HIV CAR-T 细胞，这可能是 HIV 功能性治愈的有力治疗候选物。
