Figure 4

Sensitivity vector metrics reflect biologically relevant differences in T cell subset tropism. (A) Total PBMCs were infected with luciferase reporter pseudotypes bearing wt, S142N, or K421D JR-CSF envelopes. VSV-G pseudotypes were used as positive controls. All infections (except for VSV-G) could be inhibited by maraviroc (>95%). Error bars represent ranges between two experiments. (B) Scheme for using CCR7 (PE-Cy7) and CD45RO (FITC) to identify the following T-cell subsets: Naïve (CCR7+ CD45RO-), Central Memory (T_CM, CCR7+ CD45RO+), Effector Memory (T_EM, CCR7- CD45RO+), and Effector Memory RA (T_EMRA, CCR7- CD45RO-). (C) and (D) CD8-depleted PBMCs were infected with the indicated pseudotyped viruses at an MOI of 20 (as titered on Ghost-R5 cells). Three days post-infection, cells were analyzed by multi-color flow cytometry. (C) Infected cells were identified by intracellular p24 staining using PE-conjugated KC57 Mab. (D) Uninfected T-cell subset distribution is shown in grey density plot, while infected p24+ cells are overlaid as the red dots. The percent of total p24+ cells are indicated in each quadrant. All infections could be inhibited by maraviroc (>90%). Data shown here is a representative of two independent donors.