IC.3. Specific Aims:
Specific Aim1: Role of NK cells and neutrophils at different disease stages of MmuPV1 infection
Rationale: The role of innate immune cells in HPV-associated cancers has been investigated in human studies (55). However, how these cells fight against papillomavirus infection at different disease stages remains unclear (8). We recently established persistent mucosal infection in immunocompetent heterozygous (Foxn1nu/+) NU/J mice. When compared with immunocompromised homozygous siblings, fewer viruses as well as less severe disease were found at the infected tissues (Fig. 3). Interestingly, we detected more NK cells and neutrophils infiltration at infection tissues (Fig.3 and data not shown). We hypothesize NK cells and neutrophils …show more content…
All mice will be synchronized at the diestrus stage with Depo-medroxyprogesterone (Depo-Provera) 3-5 days before infection (17). Three groups of each mouse strain will be infected with 1×109 virions as measured by viral genome equivalents at three mucosal sites (the vagina, anus, and tongue) as described in our previous studies (31-33). One group without viral infection will be used as the negative control. Mice will be sacrificed at 5 days, 10 weeks and 40 weeks post infection to capture the changes at different disease stages. Splenocytes and infected tissues will be harvested to monitor changes of NK and neutrophils (67). Viral transcripts (for active viral infection) and viral copy numbers (viral replication) will be monitored in the vaginal, anal, and oral lavages by real time PCR …show more content…
More severe diseases and higher viral titers will be detected in the homozygotes than in heterozygotes. We also expect to detect more NK cells and neutrophils at the infected tissues of heterozygous than those of homozygous mice. Depletion of either NK cells or neutrophils will promote higher viral infections and more advanced diseases in all the mucosal sites. We also expect to see anatomic differences at the three mucosal sites. Vaginal tissues will show the highest infection and the anal tissue will be the lowest according to our unpublished
Specific Aim1: Role of NK cells and neutrophils at different disease stages of MmuPV1 infection
Rationale: The role of innate immune cells in HPV-associated cancers has been investigated in human studies (55). However, how these cells fight against papillomavirus infection at different disease stages remains unclear (8). We recently established persistent mucosal infection in immunocompetent heterozygous (Foxn1nu/+) NU/J mice. When compared with immunocompromised homozygous siblings, fewer viruses as well as less severe disease were found at the infected tissues (Fig. 3). Interestingly, we detected more NK cells and neutrophils infiltration at infection tissues (Fig.3 and data not shown). We hypothesize NK cells and neutrophils …show more content…
All mice will be synchronized at the diestrus stage with Depo-medroxyprogesterone (Depo-Provera) 3-5 days before infection (17). Three groups of each mouse strain will be infected with 1×109 virions as measured by viral genome equivalents at three mucosal sites (the vagina, anus, and tongue) as described in our previous studies (31-33). One group without viral infection will be used as the negative control. Mice will be sacrificed at 5 days, 10 weeks and 40 weeks post infection to capture the changes at different disease stages. Splenocytes and infected tissues will be harvested to monitor changes of NK and neutrophils (67). Viral transcripts (for active viral infection) and viral copy numbers (viral replication) will be monitored in the vaginal, anal, and oral lavages by real time PCR …show more content…
More severe diseases and higher viral titers will be detected in the homozygotes than in heterozygotes. We also expect to detect more NK cells and neutrophils at the infected tissues of heterozygous than those of homozygous mice. Depletion of either NK cells or neutrophils will promote higher viral infections and more advanced diseases in all the mucosal sites. We also expect to see anatomic differences at the three mucosal sites. Vaginal tissues will show the highest infection and the anal tissue will be the lowest according to our unpublished