HIV-1 infection renders brain vascular pericytes susceptible to the extracellular glutamate
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Reduced pericytes’ coverage of endothelium in the brain is one of the structural changes leading to breach of the blood-brain barrier during HIV infection. We previously showed in central memory T (TCM) cells that HIV latency increases cellular susceptibility to DNA damage. In this study, we investigated susceptibility of primary brain pericytes infected with HIV-1 to DNA damage in response to glutamate and TNF-α, both known to induce neuronal death during chronic inflammatory conditions. To infect pericytes, we used a single-cycle HIV-1 pseudotyped with VSV-G envelope glycoprotein and maintained the cultures until latency was established. Our data indicate that pericytes silence HIV-1 expression at similar rate compared to primary TCM cells. TNF-α and IL-1β caused partial reactivation of the virus suggesting that progression of disease and neuroinflammation might facilitate virus reactivation from latency. Significant increases in the level of γH2AX, which reflect DNA damage, were observed in infected cultures exposed to TNF-α and glutamate at day 2 post-infection. Glutamate, an excitatory neurologic stimuli, also caused increases in the γH2AX level in latently infected pericytes, whereas PARP and DNA-PK inhibitors caused reductions in cell population suggesting that HIV-1 latency affects repairs of single- and double-strand DNA breaks. For comparison, we also analyzed latently infected astrocytes and determined that DNA damage response in astrocytes is less affected by HIV-1. In conclusion, our results indicate that productive infection and HIV-1 latency in pericytes interfere with DNA damage response, rendering them vulnerable to the agents that are characteristic of chronic neuroinflammatory disease conditions.
KeywordsPericytes Astrocytes Central memory T cells HIV latency Glutamate DNA damage response
We would like to thank Dr. Vicente Planelles for providing the DHIV construct. We would like to thank Dr. Meera Singh, Dr. Vir Singh, Dr. Sumanun Noina Suwunnakorn, Emily Weber, Sydney Simpson, and Jacob Botros-Greenlee for editing the manuscript and for their valuable comments.
This research was supported by the National Institutes of Health (NIH) grant R21 AI131961 to D.P.P. and also NIH grants R01 NS066801 and R01 NS054578 to S.B.M. The work was also supported by the University of Rochester Center for AIDS Research (NIH P30 AI078498). Funding for open access charge: National Institutes of Health.
Compliance with ethical standards
The Research Subjects Review Board at the University of Rochester Medical Center approved studies involving human samples. All the study participants were adults and blood samples were obtained after written informed consent, in accordance with the Declaration of Helsinki.
Conflict of interest
The authors declare that they have no conflict of interest.
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