Fig. 5: RhCMV encoded vFcγRs antagonize the clearance of viral genomes from plasma in vivo. | Nature Communications

Fig. 5: RhCMV encoded vFcγRs antagonize the clearance of viral genomes from plasma in vivo.

From: Rhesus Cytomegalovirus-encoded Fcγ-binding glycoproteins facilitate viral evasion from IgG-mediated humoral immunity

Fig. 5: RhCMV encoded vFcγRs antagonize the clearance of viral genomes from plasma in vivo.

A Four RhCMV-seronegative male RM were inoculated with an FL-RhCMVΔΔΔ. Three RhCMV-seronegative male RM and 12 RhCMV-seronegative female RM were inoculated with FL-RhCMV or FL-RhCMV plus RhCMV UCD52 respectively. DNA was isolated form plasma samples, and viral genome copy numbers were determined by qPCR using primer/probe sets targeting exon 1 of the IE locus. Copy numbers are reported as mean +/− SD via standard curve interpolation. B Kaplan-Meyer survival analysis showing time to control of plasma DNAemia (first-time point with genome copy numbers below the detection limit (1 copy/well). The median time to control was 31.5 days post-infection in animals infected with vFcγR-deleted RhCMV compared to 59.5 days in animals infected with FL-RhCMV (p = 0.031). C IgG binding to whole FL-RhCMV virions, RhCMV gB, and RhCMV pentamer was measured by ELISA and is shown as mean +/− SD. D ADCP is reported as the percentage of live THP-1 cells containing fluorescently labeled FL-RhCMV after incubation with plasma. ADCC is reported as the percentage of live rhesus CD16-expressing NK92 cells expressing the degranulation marker CD107a following co-incubation with FL-RhCMV infected fibroblasts and plasma samples. All responses are shown as mean +/− SD and demonstrate comparable magnitude and kinetics between animals infected with FL-RhCMV (n = 3) versus vFcγR-deleted RhCMVΔΔΔ (n = 4). E Kinetics of viral genome copy numbers in the plasma of CD4 + T cell-depleted RhCMV-naïve (seronegative) animals infected with vFcγR-deleted RhCMV (red) versus FL-RhCMV (black). Averages from immunocompetent RhCMV-seronegative RM infection in (A) are shown for comparison (gray). F Two RhCMV-seropositive RMs were inoculated with 5 × 106 PFU of vFcγR-deleted RhCMVΔΔΔ carrying an SIV-5’pol transgene as an immunological marker replacing the Rh13.1 ORF. The onset of SIV 5’Pol-specific CD4+ and CD8 + T cell responses and the boosting of RhCMV-specific T cell responses were measured in peripheral blood by intracellular cytokine staining (ICS) for IFN-γ and TNF-α using either a pool of 15mer peptides overlapping by 11 amino acids (AA) corresponding to SIV-5’pol52 or RhCMV lysates. The frequency of IFN-γ + and/or TNF-α + memory T cells is shown for each individual animal, and each indicated time point post-inoculation. Gating Strategies in Fig. S10. Source data are provided as a Source Data file.

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