FIG 6.

Pseudotyping of HIV-1 ΔNef particles with VSV-G prevents SERINC5-mediated proinflammatory cytokine production by MDMs. (A, upper panel) Relative infectivity of HIV-1 ΔNef particles pseudotyped with VSV-G produced in 293T cells analyzed as described in the legend to Fig. 1A. (Lower panel) Representative Western blot analysis of these virions for incorporation of SERINC5.HA, Nef, and HIV-1 p24. (B) Schematic of experimental flow (as in Fig. 2B, but including additional infections of MDMs with VSV-G HIV-1). (C) Quantification of intracellular p24 levels in MDMs relative to cultures infected with ΔNef particles (set to 100%, with the percentage of p24⁺ cells ranging from 1% to 6% between donors). Shown are data points from cells of individual donors, with the mean of all donors indicated by a black line. (D) Cytokine production of MDMs infected with HIV-1 Env or VSV-G containing ΔNef+S5 particles relative to the corresponding ΔNef control. Data are displayed as log₂ fold change, with red, black, and blue, respectively, indicating upregulated, unaltered, or reduced cytokine production. (E) Quantification of IL-6 concentration in the supernatant of the MDMs analyzed in panels C and D relative to the corresponding ΔNef (set to 1). Shown are data points from cells of individual donors, with the mean of all donors indicated by a black line. (F and G) Pearson’s correlation between the ΔNef/ΔNef+S5 infectivity ratio and induction of IL-6 production by ΔNef+S5 particles containing HIV-1 Env (F) or VSV-G (G). Statistics (Student's t test): n.s., nonsignificant; **, P < 0.01.