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. 2018 Nov 23;9:2719. doi: 10.3389/fimmu.2018.02719

Figure 1.

Figure 1

Myeloid cells in human cerival explants take up HIV. (A) Scheme for sorting immune cells from human cervical tissue. The figure shows 10,000 events recorded before cell sorting to set gates in a representative single cell suspension from a human cervical explant. The CD45+ cell gate (1) excluded CD45- and Sytox blue-stained dead cells. The mononuclear cell gate (2) based on forward and side scatter. CD3+ T cells (3) and CD14+ myeloid cells (4). CD3+ T cells from gate 3 separated into CD4+ (5) and CD8+ (6) T cells. CD14+ myeloid cells from gate 4 separated into CD11c– (7) macrophages and CD11c+ dendritic cells (8). (B) HIV RNA in cervical immune cells. HIV infection, assessed by qRT-PCR for gag, relative to GAPDH, in CD3+ CD4+ and CD8+ T cells and CD14+ myeloid cells, sorted from human cervical explants from five donors 20 h post-infection with HIV strain JRCSF and in uninfected control tissues. The lines indicate mean and SEM. (C) CD4 expression, measured by qRT-PCR, in sorted cervical CD4+ T cells, CD14+ myeloid cells, and CD8 T cells (representative data from donor 3). (D) Comparison of HIV RNA levels, assayed by gag qRT-PCR relative to GAPDH, in sorted cervical CD3+CD4+ T cells and CD14+ myeloid cells sorted from five donors 20 h post-infection with JRCSF or JRCSF packaged with Vpx protein. (E) HIV gag RNA by qRT-PCR in CD3+CD4+ T cells, CD14+ myeloid cells and CD3+CD8+ T cells sorted from uninfected or infected human cervical explants from donor 4, 5 days post-HIV infection (p.i.) and two additional cervical explants tested 7 days p.i. qRT-PCR values below the level of detection were arbitrarily assigned the value 0.00001. *p < 0.05, **p < 0.01, ***p < 0.001, paired t-test.