Owing to the divergent results between HPV16 and MusPV1 obtained with heparin inhibition in vivo and with binding to ECM, we examined the inhibition of in vitro infection with heparin over a range of concentrations (Fig. 7). Heparin treatment completely inhibited MusPV1 infection of HaCaT cells, whereas Pam212 infection was only inhibited 60% at the highest concentration of heparin, at over 109 M excess of heparin/VLP. We also examined the ability of exogenous heparin to prevent infection of two other murine T-5224 lines, 308 and S1. MusPV1 infection was also poorly inhibited in these lines. Complete abrogation was not attainable even at the highest concentration of heparin, similar to that seen with MusPV1 infection of the Pam212 cell line. In contrast, HPV16 PsV infection of the three murine keratinocyte-derived cell lines was completely inhibited by heparin treatment. We did not obtain complete inhibition of HaCaT cell infection by HPV16 in zone of elongation experiment, although in previous assays, both published and unpublished, complete inhibition has been observed at high concentrations (Fig. 7) (Drobni et al., 2003 and Johnson et al., 2009).
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