PT  - JOURNAL ARTICLE
AU  - Hanyu Pan
AU  - Jing Wang
AU  - Huitong Liang
AU  - Zhengtao Jiang
AU  - Lin Zhao
AU  - Yanan Wang
AU  - Xinyi Yang
AU  - Zhiming Liang
AU  - Xiaoting Shen
AU  - Qinru Lin
AU  - Yue Liang
AU  - Jinglong Yang
AU  - Panpan Lu
AU  - Yuqi Zhu
AU  - Weirong Xiang
AU  - Min Li
AU  - Pengfei Wang
AU  - Huanzhang Zhu
TI  - Allogeneic gene-edited HIV-specific CAR-T cells secreting PD-1 blocking scFv enhance anti-HIV activity in vitro
AID  - 10.1101/2022.01.14.476413
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.01.14.476413
4099  - http://biorxiv.org/content/early/2022/01/15/2022.01.14.476413.short
4100  - http://biorxiv.org/content/early/2022/01/15/2022.01.14.476413.full
AB  - HIV-specific chimeric antigen receptor (CAR) T cells have been developed to target latently infected CD4+ T cells that express virus either spontaneously or after intentional latency reversal. However, the T-cell exhaustion and the patient-specific autologous paradigm of CAR-T hurdled the clinical application. Here, we created HIV-specific CAR-T cells using human peripheral blood mononuclear cells and a 3BNC117-E27 CAR (3BE CAR) construct that enables the expression of PD-1 blocking scFv E27 and the single-chain variable fragment of the HIV-1-specific broadly neutralizing antibody 3BNC117 to target native HIV envelope glycoprotein (Env). In comparison with T cells expressing 3BNC117-CAR alone, 3BE CAR-T cells showed greater anti-HIV potency with stronger proliferation capability, higher killing efficiency (up to ~75%) and enhanced cytokine secretion in the presence of HIV envelope glycoprotein-expressing cells. Furthermore, our approach achieved high levels (over 97%) of the TCR-deficient 3BE CAR-T cells with the functional inactivation of endogenous TCR to avoid graft-versus-host disease without compromising their antiviral activity relative to standard anti-HIV CAR-T cells. These data suggest that we have provided a feasible approach to large-scale generation of “off-the-shelf” anti-HIV CAR-T cells in combination with antibody therapy of PD-1 blockade, which can be a powerful therapeutic candidate for the functional cure of HIV.Competing Interest StatementThe authors have declared no competing interest.