The pentameric complex is not required for vertical transmission of cytomegalovirus in seronegative pregnant rhesus macaques

Wang, H.-Y.; Taher, H.; Kreklywich, C. N.; Schmidt, K. A.; Scheef, E. A.; Barfield, R.; Otero, C. E.; Valencia, S.; Crooks, C. M.; Mirza, A.; Woods, K.; Burgt, N. V.; Kowalik, T. F.; Barry, P. A.; Hansen, S. G.; Tarantal, A. F.; Chan, C.; Streblow, D. N.; Picker, L. J.; Kaur, A.; Frueh, K.; Permar, S. R.; Malouli, D.

doi:10.1101/2023.06.15.545169

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bioRxiv · 10.1101/2023.06.15.545169

The pentameric complex is not required for vertical transmission of cytomegalovirus in seronegative pregnant rhesus macaques

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Abstract

Congenital cytomegalovirus (cCMV) infection is the leading infectious cause of neonatal neurological impairment but essential virological determinants of transplacental CMV transmission remain unclear. The pentameric complex (PC), composed of five subunits, glycoproteins H (gH), gL, UL128, UL130, and UL131A, is essential for efficient entry into non-fibroblast cells in vitro. Based on this role in cell tropism, the PC is considered a possible target for CMV vaccines and immunotherapies to prevent cCMV. To determine the role of the PC in transplacental CMV transmission in a non-human primate model of cCMV, we constructed a PC-deficient rhesus CMV (RhCMV) by deleting the homologues of the HCMV PC subunits UL128 and UL130 and compared congenital transmission to PC-intact RhCMV in CD4+ T cell-depleted or immunocompetent RhCMV-seronegative, pregnant rhesus macaques (RM). Surprisingly, we found that the transplacental transmission rate was similar for PC-intact and PC-deleted RhCMV based on viral genomic DNA detection in amniotic fluid. Moreover, PC-deleted and PC-intact RhCMV acute infection led to similar peak maternal plasma viremia. However, there was less viral shedding in maternal urine and saliva and less viral dissemination in fetal tissues in the PC-deleted group. As expected, dams inoculated with PC-deleted RhCMV demonstrated lower plasma IgG binding to PC-intact RhCMV virions and soluble PC, as well as reduced neutralization of PC-dependent entry of the PC-intact RhCMV isolate UCD52 into epithelial cells. In contrast, binding to gH expressed on the cell surface and neutralization of entry into fibroblasts by the PC-intact RhCMV was higher for dams infected with PC-deleted RhCMV compared to those infected with PC-intact RhCMV. Our data demonstrates that the PC is dispensable for transplacental CMV infection in our non-human primate model. One Sentence SummaryCongenital CMV transmission frequency in seronegative rhesus macaques is not affected by the deletion of the viral pentameric complex.

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Wang, H.-Y., Taher, H., Kreklywich, C. N., Schmidt, K. A., Scheef, E. A., Barfield, R., Otero, C. E., Valencia, S., Crooks, C. M., Mirza, A., Woods, K., Burgt, N. V., Kowalik, T. F., Barry, P. A., Hansen, S. G., Tarantal, A. F., Chan, C., Streblow, D. N., Picker, L. J., Kaur, A., Frueh, K., Permar, S. R., Malouli, D.. 2023-06-16. The pentameric complex is not required for vertical transmission of cytomegalovirus in seronegative pregnant rhesus macaques. https://doi.org/10.1101/2023.06.15.545169

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