Tracking vaginal, anal and oral infection in a mouse papillomavirus infection model

Abstract

Noninvasive and practical techniques to longitudinally track viral infection are sought after in clinical practice. We report a proof-of-principle study to monitor the viral DNA copy number using a newly established mouse papillomavirus (MmuPV1) mucosal infection model. We hypothesized that viral presence could be identified and quantified by collecting lavage samples from cervicovaginal, anal and oral sites. Nude mice infected at these sites with infectious MmuPV1 were tracked for up to 23 weeks starting at 6 weeks post-infection. Viral DNA copy number was determined by SYBR Green Q-PCR analysis. In addition, we tracked viral DNA load through three complete oestrous cycles to pinpoint whether there was a correlation between the DNA load and the four stages of the oestrous cycle. Our results showed that high viral DNA copy number was reproducibly detected from both anal and cervicovaginal lavage samples. The infection and disease progression were further confirmed by histology, cytology, in situ hybridization, immunohistochemistry and transmission electron microscopy. Interestingly, the viral copy number fluctuated over the oestrous cycle, with the highest level at the oestrus stage, implying that multiple sampling might be necessary to provide a reliable diagnosis. Virus DNA was detected in oral lavage samples at a later time after infection. Lower viral DNA load was found in oral samples when compared with those in anal and vaginal tracts. To our knowledge, our study is the first in vivo study to sequentially monitor papillomavirus infection from mucosal anal, oral and vaginal tracts in a preclinical model.

Spontaneous and Vaccine-Induced Clearance of Mus Musculus Papillomavirus 1 Infection

“…While we studied skin disease here, MmuPV1 also replicates at oral, anal, and vaginal sites, suggesting how this model might be further improved (80). Using mice with persistent benign MmuPV1 papillomas, we demonstrate immune-directed clearance through vaccination with a naked DNA, hCRT-mE6mE7mL2, administered with electroporation.…”

Papillomavirus can be transmitted through the blood and produce infections in blood recipients: Evidence from two animal models

“…Conceptrol (Ortho Options, over the counter) as previously described . Tongues 519 were withdrawn using a sterile forceps and microneedles were used to wound the ventral surface 520 of the tongues (Cladel et al, 2016;Hu et al, 2015). Twenty-four hours after wounding, eight 521 female and six male Hsd: Nu athymic mice were again anesthetized and challenged with 522 infectious MmuPV1 virions (1×10 8 viral DNA equivalents) via the tail vein.…”

Recent advances in preclinical model systems for papillomaviruses

“…The virus can infect both cutaneous and mucosal epithelium (Cladel, Budgeon et al, 2013a; Cladel, Budgeon et al, 2015a;Hu, Budgeon et al, 2015)…”

“…Experiments have shown that back, muzzle and tail skin are susceptible sites and led the initial investigators to conclude that this virus shows a cutaneous tropism (Ingle, Ghim et al, 2011). However, additional studies demonstrated that secondary infections could be found in mucosal tissues of the vaginal, anal and oral cavities (Cladel, Budgeon et al, 2013a;Cladel, Budgeon et al, 2015b) and that infections could be experimentally induced at these sites with ease (Cladel, Budgeon et al, 2015a; Hu, Budgeon et al, 2015). More recent studies in our lab with several strains of mice with varying levels of immune deficiency have indicated that there are unknown mechanisms of altered tissue tropism in which vaginal sites in two strains show active and persistent infections whereas direct infection of tail skin showed no signs of clinical disease.…”