(FIG. 1-2-3-4) Telomere and ATM dynamics in CD4 T cell depletion in active and virus-suppressed HIV-1 infection

Published: 24-09-2019| Version 1 | DOI: 10.17632/5mhgx2t9c3.1
Contributors:
Sushant Khanal,
Zhi Qiang Yao

Description

Fig.1. HIV-1 infection of SupT1 cells. A) Bright and fluorescent microscopic examinations of HEK293T cells transfected for 48 h with mock or HIV-1 NL4-3 mCherry plasmid. B) Flow cytometry analysis of p24 expression in SupT1 cells with HIV-1 infection. C) RT-PCR detection of HIV-1 gene expression in SupT1 cells with HIV infection and RAL treatment (added 2 h after HIV infection). D) Flow cytometry analysis of p24 expression in SupT1 cells with HIV infection and RAL treatment (added 24 h after HIV infection). E) Time-dependent p24 expression in SupT1 cells with HIV infection and RAL treatment. F) RT-PCR detection of HIV-1 gene expression in SupT1 cells with HIV infection and RAL treatment (added 24 h after HIV infection). Fig.2. HIV-1 infection induces SupT1 cell apoptotic death, which is reversible by the antiviral treatment. A- B) Representative dot plots and summary data of flow cytometry analysis of early (Av+ 7AAD-) and late (Av+ 7AAD+) apoptosis of SupT1 cells with HIV infection and RAL treatment (added 2 h after HIV infection). C-E) Representative dot plots and summary data of flow cytometry analysis of early (Av+ 7AAD-) and late (Av+ 7AAD+) apoptosis of SupT1 cells with HIV infection and RAL treatment (added 24 h after HIV infection). F) Manual count of the number of CD4 T cells with HIV-1 infection and RAL treatment. Fig.3. HIV-1 infection of primary CD4 T cells induces apoptosis, which was reversed by the RAL treatment. A-B) Flow cytometry analysis of p24 expression in PHA-activated CD4+, CD8+, and CD19+ lymphocytes with or without HIV-1 infection at the indicated time points (n=3). C) p24 expression in different subsets of anti-CD3/CD28-activated CD4+ T cells with HIV-1 infection at indicated times. D) p24 expression in TCR-activated CD4 T cells with HIV infection and RAL treatment. E) Representative dot plots and summary data of flow cytometry analysis (n=6) of early (Av+ 7AAD-) and late (Av+ 7AAD+) apoptosis of primary CD4 T cells with HIV infection and RAL treatment (added 24 h after HIV infection). Fig.4. The extrinsic death pathways are not involved in HIV-induced CD4 T cell apoptosis in vitro. A) Av and 7AAD expression on SupT1 cells with HIV-1 infection and RAL treatment in the presence of anti-CD178, anti-CD40 and anti-CD253 blocking or isotype control antibodies. B) Av and 7AAD expression on primary CD4 T cells with or without HIV-1 infection and RAL treatment, in the presence of anti-CD178, anti-CD40 and anti-CD253 blocking or isotype control antibodies for 5 days (n=3).

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