Glycan dependent phenotype differences of HIV-1 generated from macrophage versus CD4+ T helper cell populations

Abstract

Human immunodeficiency virus type 1 (HIV-1) is able to infect a variety of cell types with differences in entry efficiency and replication kinetics determined by the host cell type or the viral phenotype. The phenotype of the virus produced from these various cell types, including infectivity, co-receptor usage and neutralisation sensitivity, may also be affected by the characteristics of the producing cell. This can be due to incorporation of variant cell-specific molecules or differences in post-translational modifications of the gp41/120 envelope. In this study we produced genetically identical virus strains from macrophages, CD4-enriched lymphocytes as well as Th1 and Th2 CD4+ cell lines and compared each different virus stock for their infectivity in various cell types and sensitivity to neutralisation. In order to study the effect of the producer host cell on the virus phenotype, virus stocks were normalised on infectivity and were sequenced to confirm env gene homogeneity. Virus production by Th1 or Th2 cells did not compromise infectivity of the variant cell types tested. We observed no difference in sensitivity to co-receptor blocking agents upon viral passage through Th1 and Th2 CD4+ cell lineages nor did this affect DC-SIGN-mediated viral capture as measured in a transfer assay to CD4+ lymphocytes. Virus produced by macrophages was comparably sensitive to CC-chemokine inhibition as was virus generated from the array of CD4+ lymphocytes. We identified that virus produced from macrophages was fourteen times more resistant to 2G12 neutralisation than virus produced from CD4+ lymphocytes. Macrophage-produced dual-tropic (R5/X4) virus was six times more efficiently transmitted to CD4+ cells than lymphocyte-derived HIV-1 (p<0.0001) after DCSIGN capture. These results provide further insights to what extent the host cell influences viral phenotype and thereby various aspects of HIV-1 pathogenesis but suggest that viruses generated from Th1 versus Th2 cells are consistent in phenotype.

Macrophage- and CD4+T cell-derived SIV differ in glycosylation, infectivity and neutralization sensitivity

“…M-SIV was more efficiently transferred to target cells by DC-SIGN compared to T-SIV (t-test, p = 0.01). This observation is consistent with previous findings related to the transmission of dual-tropic HIV produced in macrophages versus CD4 + T cells through DC-SIGN (Heeregrave et al 2023). Similarly, DC-SIGNR and Langerin preferentially transferred M-SIV in comparison to T-SIV (t-test, p = 0.0001, 0.0003).…”

“…3B). These results underscore that the infectivity of M-SIV per ng capsid antigen surpasses that of T-SIV, confirming the findings previously obtained for SIV (Gaskill et al 2008) and dual-tropic HIV (Heeregrave et al 2023).…”

“…Our results support the conclusions of others that cell type-dependent differences in HIV and SIV, like Env glycosylation, influence the sensitivity towards potential biological interventions including lectins and antibodies (Heeregrave et al 2023;Raska et al 2010;Willey et al 1996). In contrast to our results, two other studies determined macrophage-derived HIV to be more neutralization sensitive than viruses produced in CD4 + T cells (Heeregrave et al 2023;Willey et al 1996). While one study utilized HIV and 2G12, a mannose-only binding neutralizing antibody, for their investigations (Heeregrave et al 2023), the other study made this conclusion by using sera of an HIV infected chimpanzee (Willey et al 1996).…”

“…Our results support the conclusions of others that cell type-dependent differences in HIV and SIV, like Env glycosylation, influence the sensitivity towards potential biological interventions including lectins and antibodies (Heeregrave et al 2023; Raska et al 2010; Willey et al 1996). In contrast to our results, two other studies determined macrophage-derived HIV to be more neutralization sensitive than viruses produced in CD4 + T cells (Heeregrave et al 2023; Willey et al 1996).…”

“…Notably, glycosylation of Env and cellular proteins is a cell type-dependent process (Liedtke et al 1994; Liedtke et al 1997; Raska et al 2010; Willey et al 1996) and differences in N -glycosylation of HIV and SIV from macrophages and CD4 + T cells have been associated with differential infectivity (Gaskill et al 2008; Heeregrave et al 2023), neutralization sensitivity (Heeregrave et al 2023; Willey et al 1996) and lectin reactivity (Heeregrave et al 2023; Lin et al 2003). Despite their importance in virus-host cell interactions and immune control, host cell-specific glycosylation differences have not been determined for HIV or SIV at a molecular level.…”