Fig 3.

Facile use of CCR5 domains by adapted HIV-1. (A) HIV-1 infections mediated by wild-type, mutant and murine CCR5s. Wild-type HIV-1_JRCSF and the variant adapted to a low amount of CCR5(HHMH) were compared for abilities to infect 293T cells transiently expressing CD4 and murine CCR5 (mu CCR5), CCR5(Δ18), CCR5(G163R) or the double mutant CCR5(Δ18, G163R). The data are means of two independent experiments performed in triplicate. Error bars are S.E.M. (B) CCR5(Δ18) and CCR5(HHMH) use by HIV-gpt viruses containing adaptive gp120 mutations. Pseudotyped viruses were used to infect HeLa-CD4 cells expressing CCR5(Δ18) (2.7 × 10⁴ molecules/cell), CCR5(HHMH)-low, or CCR5(HHMH)-high. Adaptive gp120 mutations in the virus pseudotypes were as follows: CCR5(HHMH)-Ad: S298N/F313L/N403S; CCR5(Δ18)-Ad minus N300Y: S298N/I307M/F313L/T315P/N403S; CCR5(Δ18)-Ad:S298N/N300Y/I307M/F313L/T315P/N403S. The data are from 2 independent experiments performed in duplicate. Error bars are S.E.M. (C) Infections mediated by CCR5(Δ18) plus sulfated N-terminal CCR5 peptide. HeLa-CD4 cells expressing 2.7 × 10⁴ CCR5(Δ18) molecules/cell were infected in the presence of varying concentrations of CCR5 peptide (0, 25, 100, and 200 μM), and infectivities (i_rel) were measured relative to JC.53 cells. The replication competent CCR5(Δ18)-adapted, CCR5(HHMH)-adapted, and wild-type JRCSF (blue, green, and red curves, respectively) isolates were tested. The graph shows a representative experiment performed in duplicate. Error bars are the range.