Fig 1.
Gating strategy for CD32 expression on peripheral CD4+ and CD4dimCD8bright T cell populations.
Lymphocytes were gated from whole human PBMCs using forward and side scatter area, then two singlet gates were utilized to remove doublet populations. This was followed by an Aqua Live/Dead stain and total CD3+ cell gate. From this, two different gates were determined: total CD3+CD8+ cells and CD3+ CD4 single positive cells. From the CD3+CD8+cells, CD8 single positive T cell and CD4dimCD8bright T cell populations were obtained. CD4 single positive T cells, CD4brightCD8bright, and CD8-CD4- were gated from total CD3+ cells. The results of these gates are five T cell populations: CD8+, CD4dimCD8bright, CD4+, CD4brightCD8bright, and CD8-CD4-. CD32 expression was determined through usage of a CD32 FMO, which was used as a negative control for CD32 staining and allowed for accurate gating on the positive populations only.
Fig 2.
CD32 expression on CD4+ and CD4dimCD8bright T cells among HIV seronegative and seropositive donors.
a) CD32 expression on CD4dimCD8bright T cells and CD4+ T cells from HIV+ and HIV- blood donors was evaluated by flow cytometry using peripheral blood of HIV seronegative (n = 4) and HIV seropositive (n = 11 donors). b) PBMCs from elite controllers (EC) (undetectable viral loads for >5 years) including one long term non-progressor (high viral load, CD4 > 500 cells/μL), cART adherent (viral load <40 copies/mL), and cART non-adherent/ treatment naïve (viremic) (viral load >10,000 copies/mL, CD4 count <500 cells/μL) donors were evaluated by flow cytometry for CD32 expression. c) Relationship between CD32 expression on CD4dimCD8bright T cells and CD4 count of HIV+ participants. Clinical CD4 counts taken on day of blood draw. * denotes p<0.05 determined by unpaired T-test and *** denotes p<0.01 as measured by paired T-test.
Table 1.
Clinical description of study participants.
Fig 3.
CD32 expression is higher on various CD4 and CD8 double positive T cell subsets.
a) CD32 expression was determined on five T cell populations: CD8+, CD4dimCD8bright, CD4+, CD4brightCD8bright, and CD8-CD4- cells. Positive CD32 was gated using FMO as a negative control. Expression of CD32 on each population was compared between HIV- and HIV+ donors. * denotes p<0.05 as measured by unpaired T-test. b) Within group comparisons of CD32 expression on T cell populations in HIV- and HIV+ donors was determined via ANOVA.