Figure 4.

Effects of the R5Nt moiety in the context of the trispecific CD4-MBL-R5Nt CAR. (A) Flow cytometry analysis of surface expression of CARs. After gating on live T lymphocytes, the levels of CAR expression were determined by the presence of CD4 on the CD8⁺ T cell populations (inside box). Cells were analyzed at day 11 post-transduction; inside boxes indicate % CD4-positive. (B) Activities of CARs in the HIV-1 spreading infection assay for the BX08 (left panels) and JR-FL (right panels) primary isolates. Cocultures at varying E:T ratios were continued for 19 days, with aliquots taken at 2-3-day intervals for p24 assay. Top panels show the effects of varying E:T ratios. p24 values are shown for BX08 at day 15, and for JR-Fl at day 7. In both graphs, the dotted line represents Target cells only. Bottom panels show the time courses of infection. P24 values are shown for cocultures at E:T ratio of 0.04:1. In all graphs, the dashed line represents the limit of detection determined from the standard curve. All assay points represent the mean of triplicate samples, with error bars denoting standard deviation.