FIG 3.
Higher transmission fitness of Envacute versus Envchronic viruses in multivirus competition assays in human tissue. (a) Penile tissues were dissected into 3-mm3 explants and exposed to eight different mixtures of a panel of Envacute viruses and the same three Envchronic viruses. (b) Following overnight culture, migrating cells from the tissue were collected and cocultured with PM1 T cells (MC+PM1). After 10 days in culture, DNA from tissue and MC+PM1 cells was isolated, PCR amplified, and analyzed by NGS. The cumulative percent replication in penile tissue (a) and in MC+PM1 cocultures (b) of Envchronic and Envacute viruses for tissue A and tissue B in each of the eight multivirus panels is shown. (c) The combined average replication of all Envchronic and all Envacute viruses of the eight virus competitions in penile tissues and Envchronic (green bars) and Envacute (red bars) viruses in MC+PM1 cocultures was determined. Each competition was done in triplicates in tissues from two donors. (d) Average replication of Envchronic (bars with green outlines) and Envacute (bars with red outlines) viruses of one competition set in cervical tissue and the corresponding MC+PM1 cocultures were evaluated by NGS. Statistical significance was determined using the Holm-Sidak method (a and b) (*, P < 0.05) and a two-tailed Mann-Whitney test (c and d) (***, P < 0.001; ****, P < 0.0001).