Human Herpesvirus-6A/B Binds to Spermatozoa Acrosome and Is the Most Prevalent Herpesvirus in Semen from Sperm Donors
Figure 3
Identification of HHV-6B binding to sperm was assessed by flow cytometry, analyzing 50.000 sperm per sample. (A) Thirty straw of semen were pooled and purified with PureSperm prior to incubation with HHV-6B. After incubation the sperm were washed twice in PBS/2%FCS and stained for the presence of HHV-6B with anti-gp60/110 (lower two panels), and anti-IgG2b as a control for binding specificity (upper two panels). Vehicle was used as a negative control. (B) Quantitation of three experiments analyzing the concentration-dependent binding of HHV-6B to sperm. Samples were prepared as described in (A). SD is indicated on top of each bar. (C) One straw of semen was incubated with HHV-6B diluted 1∶10 and subsequently stained with anti-gp60/110 and anti-IgG2b followed by a goat anti-mouse FITC-conjugated antibody. (D) Ten semen samples identified as HHV6A/B negative (five) or HHV-6 positive (five) by the PCR array were stained with anti-gp60/110 and anti-IgG2b and analyzed by flow cytometry. (E) Plot of delta median fluorescence intensity of data shown in (D); p<0.008, Mann-Whitney test.