a, CD32neg and CD32high (FUN-2) CD4+ T cells from human PBMCs were assessed by flow cytometry for the expression of CD19, CD40 and HLA-DR, and compared to B cells (CD3− CD14−CD19+ lymphocytes). Representative flow cytometry results of per cell antigen levels on B cells (top, blue histograms) and CD32neg and CD32high CD4+ T cells (bottom, grey and red histograms, respectively) from an HIV-1− donor. b, Representative CD32b staining of PBMCs from an HIV-1+, ART-suppressed participant. PBMCs were stained with an optimized concentration of the 2B6 monoclonal anti-CD32b antibody, followed by an antibody cocktail that included the FUN-2 monoclonal pan-CD32 antibody, as described in the Methods. Shown are the 2B6 and FUN-2 fluorescence minus one (FMO) antibody cocktail-stained samples and a sample co-stained with 2B6 and FUN-2. c, d, CD32 mRNA expression levels in CD4+CD32+ subsets. c, The relative expression of CD32A and CD32B mRNA isoforms in sorted CD4+CD32int and CD4+CD32high subsets from HIV-1+, ART-suppressed participants (n = 4). d, mRNA expression of CD32A and CD32B from patient G07W1610. e, T and B cell lineage-specific mRNA transcripts in sorted CD4+CD32+ subsets from participant G07W1610. Relative mRNA expression of target genes was normalized to ATCB using the comparative Ct method. Results are mean ± s.d. of each value from each participant (n = 4; c), or from values generated from two separate experiments using samples from the same patient (d).