Exposure to HIV-1 Directly Impairs Mucosal Epithelial Barrier Integrity Allowing Microbial Translocation

Abstract

While several clinical studies have shown that HIV-1 infection is associated with increased permeability of the intestinal tract, there is very little understanding of the mechanisms underlying HIV-induced impairment of mucosal barriers. Here we demonstrate that exposure to HIV-1 can directly breach the integrity of mucosal epithelial barrier, allowing translocation of virus and bacteria. Purified primary epithelial cells (EC) isolated from female genital tract and T84 intestinal cell line were grown to form polarized, confluent monolayers and exposed to HIV-1. HIV-1 X4 and R5 tropic laboratory strains and clinical isolates were seen to reduce transepithelial resistance (TER), a measure of monolayer integrity, by 30–60% following exposure for 24 hours, without affecting viability of cells. The decrease in TER correlated with disruption of tight junction proteins (claudin 1, 2, 4, occludin and ZO-1) and increased permeability. Treatment of ECs with HIV envelope protein gp120, but not HIV tat, also resulted in impairment of barrier function. Neutralization of gp120 significantly abrogated the effect of HIV. No changes to the barrier function were observed when ECs were exposed to Env defective mutant of HIV. Significant upregulation of inflammatory cytokines, including TNF-α, were seen in both intestinal and genital epithelial cells following exposure to HIV-1. Neutralization of TNF-α reversed the reduction in TERs. The disruption in barrier functions was associated with viral and bacterial translocation across the epithelial monolayers. Collectively, our data shows that mucosal epithelial cells respond directly to envelope glycoprotein of HIV-1 by upregulating inflammatory cytokines that lead to impairment of barrier functions. The increased permeability could be responsible for small but significant crossing of mucosal epithelium by virus and bacteria present in the lumen of mucosa. This mechanism could be particularly relevant to mucosal transmission of HIV-1 as well as immune activation seen in HIV-1 infected individuals.

Author Summary

Clinical studies have shown that HIV-1 infected patients have increased intestinal permeability. In chronically infected patients that progress to AIDS, there is activation of immune cells consistent with leakage of microbes via the gut. However, the mechanism by which this occurs is not clear. Here, we show that direct exposure of intestinal and genital epithelial cells to HIV leads to breaching of the mucosal barrier and increased leakage of both bacteria and virus across the epithelium. The mechanism of this breakdown appears to be due to inflammatory factors produced by epithelial cells themselves, in response to HIV-1 exposure, that destroy the tight junctions between epithelial cells, thereby allowing microbes access to the inside of the body. Interestingly, we found that treatment of epithelial cells with just the surface glycoprotein from HIV could lead to similar breakdown of the barrier. This implies that when mucosal epithelial cells come in direct contact with large amounts of HIV-1, the virus can cross into the inside of the body and cause direct infection of target cells. The crossing of the bacteria by similar mechanism can lead to chronic inflammation and activation of immune cells of the body.