Figure 1

Prolonged ART results in an increased frequency of classical monocytes. (A) Thawed PBMCs from HIV patients were stained with antibodies recognizing cell-surface and intracellular myeloid markers. Analysis was performed by sequentially gating on live cells, singlets (FSC-A/FSC-H), non-lymphocyte (SSC-A high/FSC-A high), lineage negative (CD3^- CD19^- CD56^-), and HLA-DR and CD11c positive populations. Myeloid cells were further defined by expression of CD14 and CD16 into three subsets (CD14^hiCD16^-, CD14^dimCD16⁺, and CD14^-CD16^- cells). Frequencies of CD14^hiCD16^- classical monocytes (lower right gate), CD14^dimCD16⁺ non-classical monocytes (upper left gate), and CD11c⁺ CD14^-CD16^- mDCs (lower left gate) and in relation to the parent myeloid gate were calculated. (B) Column statistics were performed by 1-way ANOVA on patients described in Table 1. Statistical significance is denoted as *p < 0.05, **p < 0.01, and ***p < 0.001. (C) Myeloid subpopulations were measured during pre-ART to post-ART time points from thawed PBMCs of ART-treated subjects (see "pre- ART-suppressed" subjects described in Table 2). Student’s paired t-test was performed and corresponding p values are described.