Anti-HPV16 E2 Protein T-Cell Responses and Viral Control in Women with Usual Vulvar Intraepithelial Neoplasia and Their Healthy Partners

T-cell responses (proliferation, intracellular cytokine synthesis and IFNγ ELISPOT) against human papillomavirus 16 (HPV16) E2 peptides were tested during 18 months in a longitudinal study in eight women presenting with HPV16-related usual vulvar intraepithelial neoplasia (VIN) and their healthy male partners. In six women, anti-E2 proliferative responses and cytokine production (single IFNγ and/or dual IFNγ/IL2 and/or single IL2) by CD4+ T lymphocytes became detectable after treating and healing of the usual VIN. In the women presenting with persistent lesions despite therapy, no proliferation was observed. Anti-E2 proliferative responses were also observed with dual IFNγ/IL2 production by CD4+ T-cells in six male partners who did not exhibit any genital HPV-related diseases. Ex vivo IFNγ ELISPOT showed numerous effector T-cells producing IFNγ after stimulation by a dominant E2 peptide in all men and women. Since the E2 protein is absent from the viral particles but is required for viral DNA replication, these results suggest a recent infection with replicative HPV16 in male partners. The presence of polyfunctional anti-E2 T-cell responses in the blood of asymptomatic men unambiguously establishes HPV infection even without detectable lesions. These results, despite the small size of the studied group, provide an argument in favor of prophylactic HPV vaccination of young men in order to prevent HPV16 infection and viral transmission from men to women

Epidemiological and economic burden of potentially HPV-related cancers in France.

Human papillomaviruses (HPV) infection is now known to be responsible for almost all cervical cancers, and for a substantial fraction of Head and Neck cancers (HNCs). However, comprehensive epidemiological and economic data is lacking in France, especially for rarer potentially HPV-related cancers, which include anal, vulvar and vaginal cancers. Using the national comprehensive database of French public and private hospital information (PMSI), we assessed prevalence and incidence of patients with in-hospital diagnosis for potentially HPV-related cancers in 2013, and estimated costs related to their management over a 3-year period after diagnosis in France. Concerning female genital cancers, 7,597, 1,491 and 748 women were hospitalized for cervical, vulvar and vaginal cancer in 2013, respectively, with 3,120, 522 and 323 of them being new cases. A total of 4,153 patients were hospitalized for anal cancer in 2013, including 1,661 new cases. For HNCs, 8,794 and 14,730 patients were hospitalized for oral and oropharyngeal cancer in 2013, respectively; 3,619 and 6,808 were new cases. Within the 3 years after cancer diagnosis, the average cost of hospital care per patient varied from €28 K for anal cancer to €41 K for oral cancer. Most expenditures were related to hospital care, before outpatient care and disability allowance; they were concentrated in the first year of care. The total economic burden associated with HPV-potentially related cancers was about €511 M for the French National Health Insurance over a 3 years period (2011 to 2013), ranging from €8 M for vaginal cancer to €222 M for oropharyngeal cancer. This study reported the most up-to-date epidemiological and economic data on potentially HPV-related cancers in France. These results may be used to evaluate the potential impact of new preventive strategies, namely the generalized organized screening of cervical cancer and the nine-valent HPV vaccine, indicated in the prevention of cervical, vaginal, vulvar and anal cancers

Clinical and biological characteristics of the eight women having usual VIN and their male partners.

<p>NA: not applicable.</p

Anti-HPV16 E2 peptide proliferative responses in women.

<p>T-cell proliferation induced by E2 peptide pools were studied with PBMCs from A: women presenting with usual VIN either during the whole study (F#1 and F#7) or at study entry and up to the healing after treatment (F#3, F#6, F#8 before M6, F#5 before M12) and B: asymptomatic women having cleared their usual VIN after treatment (F#2, F#4 before their entry in the study, F#3, F#6, F#8 from M6, F#5 from M12). SI is represented by the cpm in peptide-stimulated cells/cpm in negative control wells (without peptide). Each bar represents SI against a pool of E2 peptide or positive control (+) obtained from PBMCs sampled at M0 (blue), M6 (red), M12 (yellow) and M18 (turquoise blue). Proliferative responses with SI >3 were scored as positive, provided that cpm in the negative control was above 500. The values of positive SI for E2 pools are indicated in black near the corresponding bar. Responses to 15 mer peptides comprised in the various peptide pools are shown, superposed to the response to the pool, in another color. The SI values for these peptides are indicated using the same colors near the corresponding bars.</p

<i>Ex vivo</i> anti-HPV16-E2 peptide IFNγ ELISPOT responses in women presenting with usual VIN.

<p>IFNγ ELISPOT response to E2 peptide pools was studied using PBMCs from A: women presenting with usual VIN either during the whole study (F#1 and F#7) or at study entry and up to the healing after treatment (F#3, F#6, F#8 before M6, F#5 before M12) and B: asymptomatic women having cleared their usual VIN after treatment (F#2, F#4 before their entry in the study, F#3, F#6, F#8 from M6, F#5 from M12). Bars represent numbers of SFC/10⁶ PBMCs against a pool of E2 peptides, negative (0) and positive (+) controls obtained at M0 (blue), M6 (red), M12 (yellow) and M18 (turquoise blue). Standard deviation of triplicates appears for each bar. Responses were considered significant when the mean number of SFC per 10⁶ cells in the 3 experimental wells was >3-fold the mean number of SFC in the negative control wells (PBMC alone) and >30 SFC/10⁶ cells. Positive responses are identified by a black star above the corresponding bar.</p

Ex vivo IFNγ ELISPOT responses in women and men against HPV16, HPV2, HPV27 and HPV55 E2 309–323 peptide.

<p>A: number of SFC in negative control was substracted.</p><p>B: number of SFC was <3-fold the mean number of SFC in the negative control.</p

<i>Ex vivo</i> anti-HPV16-E2 peptide IFNγ ELISPOT responses in male partners of women presenting with usual VIN.

<p>IFNγ ELISPOT responses to E2 peptide pools were studied with PBMCs from male partners of women presenting with usual VIN. Bars represent numbers of SFC/10⁶ PBMCs against a pool of E2 peptides, negative (0) and positive (+) controls obtained at M0 (blue), M6 (red), M12 (yellow) and M18 (turquoise blue). Standard deviation of triplicates appears for each bar. Responses were considered significant when the mean number of SFC per 10⁶ cells in the 3 experimental wells was >3-fold the mean number of SFC in the negative control wells (PBMC alone) and >30 SFC/10⁶ cells. Positive responses are identified by a black star above the corresponding bar.</p

Anti-HPV16 E2 peptide proliferative responses in male partners of women presenting with usual VIN.

<p>Proliferative assays were performed using PBMCs in presence of pools of E2 peptides. SI is represented by the cpm in peptide-stimulated cells/cpm in negative control wells (without peptide). Each bar represents SI against a pool of E2 peptide or positive control (+) obtained from PBMCs sampled at M0 (blue), M6 (red), M12 (yellow) and M18 (turquoise blue). Proliferative responses with SI >3 were scored as positive, provided that cpm in the negative control was above 500. The values of positive SI for E2 pools are indicated in black near the corresponding bar. Responses to 15 mer peptides comprised in the various peptide pools are shown, superposed to the response to the pool, in another color. The SI values for these peptides are indicated in the same colors near the corresponding bars.</p