Currently, thirty-seven million people are infected with human immunodeficiency virus-1 (HIV-1) worldwide. Thankfully, the development of combined antiretroviral therapy (cART) regimens has decreased mortality and significantly improved the overall quality of life for these patients. However, approximately half of all patients clinically manifest with HIV-associated neurocognitive disorder (HAND), a spectrum of cognitive, motor, and behavioral abnormalities which histologically present as non-specific gliosis, synaptodendritc damage and loss of white matter and myelin. Furthermore, the severity of white matter damage correlates with the length of ART duration. However, almost no studies have been performed to determine how the myelin sheath or the oligodendrocytes that synthesize the sheath are damaged. Thus, we hypothesized that the administration of ART contributed in part to the myelin loss in the CNS of HIV-positive patients. Previously, we have reported that the protease inhibitor class of ART drugs hampered the in vitrodifferentiation of oligodendrocytes. Given that the new US guidelines for treating HIV patients recommends anew class of drugs, the integrasestrand transferinhibitors(INSTIs)as front-line therapy, we examined if two specific INSTIs, Elvitegravir (EVG) and raltegravir (RAL), altered the survival and/or maturation of developing oligodendrocytes in vitroand in vivo. We found that treatment of oligodendrocyte precursor cells (OPCs) with EVG, but not RAL, during differentiation reduced the number of cells positive for immature oligodendrocyte marker galactosylceramide (GalC) and mature oligodendrocyte marker myelin basic protein (MBP) in vitro, as well as the synthesis of myelin proteins. However, neither EVG or RAL induced cell loss or apoptosis, as determined by cell counts and TUNEL assays, suggesting that EVG does not affect OPC viability but instead, inhibits differentiation. EVG-induced oligodendrocyte differentiation deficits could be reversed by pre-treating the cells with a drug that pharmacologically inhibits the phosphorylation of eukaryotic initiation factor 2α(eIF2α) throughthe cellular integrated stress response (ISR). Finally, in vivo,mice receiving EVG/COBI failed to remyelinate the corpus callosum during the three week recovery period following demyelination, after cuprizone treatment. Although EVG/COBI treatment by itself did not cause overt white matter loss in this brain region. Our study demonstrates that EVG, but not RAL, inhibits oligodendrocyte precursor cell differentiation both in vitroand in vivo. Furthermore, EVG may be inhibiting oligodendrocyte precursor cell differentiation though activation of the ISR. Also, we found thatthe effects of EVG on oligodendrocyte differentiation could be attenuated in vitroby inhibiting the ISR. These studies suggest that ART may contribute to cognitive impairment by inhibiting renewal and replacement of oligodendrocytes in adults or development of oligodendrocytes in children. Further, our results suggest an ISR inhibitor might attenuate the negative effect of EVG on the maturation of oligodendrocytes. Our findings also suggest that development of less toxic ART compounds and adjunctive therapies are needed to minimize the side effects of ART on the CNS.