Donor variation in in vitro HIV-1 susceptibility of monocyte-derived macrophages

Abstract

Primary human cells from different donors vary in their susceptibility to in vitro infection with HIV-1. In order to perform genetic analysis to identify host factors that affect HIV-1 susceptibility, it is important that a clear phenotype is defined. Here, we report a standardized method to study variation for in vitro HIV-1 infection in monocyte-derived macrophages (MDM) from large numbers of individuals. With this assay, HIV-1 susceptibility of MDM from 489 different donors shows more than 3 log variation and a good correlation with the 32 base pair deletion in the CCR5 co-receptor (ccr5 Delta 32 genotype) of the donors. However, in 7 of 12 donors completely resistant to infection with CCR5-using HIV-1, this was not explained by the ccr5 Delta 32 genotype, showing evidence that other host factors are likely to influence HIV-1 replication in MDM. Infections with VSV-G pseudotyped HIV-1 indeed confirmed the existence of post-entry level restrictions in MDM.

Generation of Transmitted/Founder HIV-1 Infectious Molecular Clones and Characterization of Their Replication Capacity in CD4 T Lymphocytes and Monocyte-Derived Macrophages

“…Consistent with this phenotype, REJO.c and THRO.c were more sensitive to inhibition by sCD4 than were the other T/F viruses. In agreement with earlier studies, our results also indicated that MDM from different donors exhibited considerable variation in their susceptibility to productive HIV-1 infection (7,65). While our panel of 10 clade B T/F viruses exhibited a lower MRC than prototypic mac-tropic control viruses, our analysis clearly demonstrates that as a group they are replication competent in cultures of MDM.…”

“…For each donor, the median day 6 LucR values show markedly lower productive MDM infection with T/F Env-IMC-LucR viruses, and across the seven donors the difference between the T/F versus control viruses was statistically significant, as determined by the exact two-tailed Wilcoxon matched-pairs signed-rank test (P ϭ 0.0156). Notably, although considerable variation was observed in virus replication (i.e., level of LucR expression) among the different donor MDM, a phenotype previously reported (7,8,23,29), in each case there remained a difference of 1 to 3 orders of magnitude between T/F and control viruses. This analysis revealed differences in terms of virus infectivity and replication that are similar to those observed studying full-length viruses.…”

HIV-1 Induces Telomerase Activity in Monocyte-Derived Macrophages, Possibly Safeguarding One of Its Reservoirs

“…Then, MDM were infected with HIV-1 BaL, and the infection efficiency was evaluated after 12 days by quantification of intracellular and supernatant levels of p24 antigen. In agreement with previous reports, the susceptibility to HIV infection varied greatly between different donors (9,25). Table 1 summarizes the levels of p24 antigen in culture supernatants from the 10 donors as measured by ELISA.…”

“…MDM have a strong donor-dependent variation of in vitro susceptibility to HIV infection, with differences of up to 3 orders of magnitude in virus production (9,25). Similarly, we observed a different susceptibility of MDM infection with both R5 and X4 strains.…”

HIV-1 Nef Interacts with HCV Core, Recruits TRAF2, TRAF5 and TRAF6, and Stimulates HIV-1 Replication in Macrophages

“…High donor-linked variability between monocytes/macrophages from different donors, in particular for their in vitro HIV-1 susceptibility, has been reported previously [42,43]. Although we observed variability between peripheral blood cells (PBMCs, monocytes, PBLs) from different donors for NF-κB activation, NF-κB activation was 2-fold higher in cells isolated from coinfected patients than in cells isolated from monoinfected patients.…”