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The vector of anti-HIV-1 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target HIV-1. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-HIV-1 antibody linked to 41BB and CD3ζ signaling domains. And the vector product was designed for the treatment of HIV-1 infection.
CAR Construction : 3BNC117-41BB-CD3ζ Fig.1 CAR-T cells displayed preferable immunophenotype. The subset composition of UTD/3B/3BD CAR-T cells were measured by surface expression of CD45RA and CD62L. Jiang, Z., Liang, H., Pan, H., Liang, Y., Wang, H., Yang, X., ... & Zhu, H. (2021). HIV-1-specific CAR-T cells with cell-intrinsic PD-1 checkpoint blockade enhance anti-HIV efficacy in vivo. Frontiers in Microbiology, 12. |
CAR Construction : 3BNC117-41BB-CD3ζ Fig.2 CAR-T cells displayed preferable proliferation. CAR-T cells sorted for CAR expression were incubated with LHL2/3 cells (5 × 105cells) at 1:1 ratio for 5 days, and CAR+cells were counted daily to evaluate thein vitro proliferation. Jiang, Z., Liang, H., Pan, H., Liang, Y., Wang, H., Yang, X., ... & Zhu, H. (2021). HIV-1-specific CAR-T cells with cell-intrinsic PD-1 checkpoint blockade enhance anti-HIV efficacy in vivo. Frontiers in Microbiology, 12. |
CAR Construction : 3BNC117-41BB-CD3ζ Fig.3 Cell killing assay. Direct killing of LEL6 was performed using the LDH release assay. Jiang, Z., Liang, H., Pan, H., Liang, Y., Wang, H., Yang, X., ... & Zhu, H. (2021). HIV-1-specific CAR-T cells with cell-intrinsic PD-1 checkpoint blockade enhance anti-HIV efficacy in vivo. Frontiers in Microbiology, 12. |
CAR Construction : 3BNC117-41BB-CD3ζ Fig.4 cytotoxicity assay. Direct cytotoxicity effects on Jurkat cells, as the Env negative control here, were detected Jiang, Z., Liang, H., Pan, H., Liang, Y., Wang, H., Yang, X., ... & Zhu, H. (2021). HIV-1-specific CAR-T cells with cell-intrinsic PD-1 checkpoint blockade enhance anti-HIV efficacy in vivo. Frontiers in Microbiology, 12. |
CAR Construction : 3BNC117-41BB-CD3ζ Fig.5 TNF-α, IL-2 and IFN-γ production in co-cultures. Anti-HIV CAR-T cells were co-cultured with LEL6 cells (1 × 10^4cells) at 10:1 for 24 h, and supernatants were collected for ELISA. Jiang, Z., Liang, H., Pan, H., Liang, Y., Wang, H., Yang, X., ... & Zhu, H. (2021). HIV-1-specific CAR-T cells with cell-intrinsic PD-1 checkpoint blockade enhance anti-HIV efficacy in vivo. Frontiers in Microbiology, 12. |
CAR Construction : 3BNC117-41BB-CD3ζ Fig.6 CAR-T cells eliminated LEL6 cells in vivo. CAR-T cells displayed superior anti-HIV function in an HIV NCG mouse model. Jiang, Z., Liang, H., Pan, H., Liang, Y., Wang, H., Yang, X., ... & Zhu, H. (2021). HIV-1-specific CAR-T cells with cell-intrinsic PD-1 checkpoint blockade enhance anti-HIV efficacy in vivo. Frontiers in Microbiology, 12. |
CAR Construction : Fig.7 Surface plasmon resonance analyses. SPR sensorgrams showing the bindingof bNAbs expressed as IgG1 (green), IgA1 (blue), and IgA2 (red) to YU-2 gp140 or gp120 ligands expressed as normalized response units (RU) over time. Lorin, V., Malbec, M., Eden, C., Bruel, T., Porrot, F., Seaman, M. S., ... & Mouquet, H. (2017). Broadly neutralizing antibodies suppress post-transcytosis HIV-1 infectivity. Mucosal Immunology, 10(3), 814-826. |
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