Jaana Rautava

Oral Mucosa as a Reservoir of Human Papillomavirus: Point Prevalence, Genotype Distribution, and Incident Infections Among Males in a 7-year Prospective Study

BACKGROUND In addition to the anogenital malignancies, human papillomavirus (HPV) has been linked to oropharyngeal cancer as an important risk factor in both men and women. Knowledge of oral HPV infection among males is needed to elucidate the transmission routes and potential for prevention. OBJECTIVE To assess the prevalence, genotype distribution, and incidence of oral HPV infections among healthy Finnish men followed for 7 yr. DESIGN, SETTING, AND PARTICIPANTS Oral scrapings for HPV testing were taken from 131 fathers-to-be (mean age: 28.9 yr) at baseline and at 2-mo, 6-mo, 12-mo, 24-mo, 36-mo, and 7-yr follow-up visits to detect prevalent and incident HPV infections. Purified DNA extracted from scrapings was used for HPV genotyping, with the Multimetrix kit (Progen Biotechnik, Heidelberg, Germany) detecting 24 genotypes. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS Point prevalence, genotype distribution, and incident rates of oral HPV infections. Demographic data were collected using structured questionnaires, and covariates of incident oral HPV infections were analysed using uni- and multivariate Poisson regression (for panel data). RESULTS AND LIMITATIONS The point prevalence of oral HPV infection fluctuated from 15.1% to 31.1% during the follow-up period. In total, 17 different HPV genotypes were found. At baseline, the single most frequent genotype among the HPV-positive samples was HPV16 (33.3%; 8 of 24), followed by HPV33 (12.5%) and HPV82 (12.5%). Multiple-type infections comprised 16.7% (4 of 24), HPV16 being involved in all combinations. For baseline-negative men, the mean time to the first incident infection ranged from 3.9 mo (HPV82) to 25.7 mo (HPV56). None of the demographic factors was a significant independent predictor of incident oral HPV infections in multivariate models. CONCLUSIONS Detection of oral HPV DNA carriage in men is common, HPV16 being the most prevalent genotype. Oral mucosa may play a significant role in HPV transmission.

Prevalence, genotype distribution and persistence of human papillomavirus in oral mucosa of women: a six-year follow-up study.

Background Human papillomavirus (HPV) infections have been linked to a subset of oral and oropharyngeal cancers. However, little is known on the natural history of oral HPV infections. We designed the prospective Finnish HPV Family Study to assess the dynamics of HPV infections in parents and their infants. This study reports HPV genotype distribution and virus persistence in oral mucosa of the mothers. Materials and Methods Totally, 324 pregnant women were enrolled at the 3rd trimester of pregnancy and followed-up for 6 years. Oral scrapings taken with a brush were collected and HPV-genotyping was performed with nested PCR and Multimetrix® test (Progen, Heidelberg, Germany). The predictors of persistent oral HPV species 7/9 infections were analyzed using generalized estimating equation models. Results The point prevalence of oral HPV varied from 15% to 24% during the 6-year follow-up. Altogether, 18 HPV genotypes were identified either as single or multiple-type oral infections. HPV16 was the most prevalent type at 9.7%–18.4%, followed by HPV18, HPV6, and multiple infections. Altogether, 74 women had persistent oral HPV infection determined as at least two consecutive samples positive with the same HPV genotype. HPV16 and HPV6 were the two most frequent types to persist (76% and 9%) for a mean of 18.6 and 20.2 months, respectively, followed by multiple infections (8%) for 18.3 months. An increased risk for persistent oral HPV infection with species 7/9 was associated with being seropositive for low-risk (LR)-HPV-types at baseline, whereas the use of oral contraceptives and a second pregnancy during follow-up were protective. Clinical oral lesions were detected in 17% of these women, one-third of whom had persistent oral HPV-infections. Conclusion HPV16 and HPV6 were the most common genotypes in oral HPV-infections and were also most likely to persist. Use of oral contraceptives and a second pregnancy protected against oral HPV persistence.

HPV genotypes and their prognostic significance in head and neck squamous cell carcinomas

BACKGROUND Human papillomavirus (HPV) has been reported in up to 50% of head and neck squamous cell carcinomas (HNSCCs). Presence of HPV in HNSCC has been associated with more favorable prognosis. OBJECTIVES This study was designed to disclose HPV genotype distribution in head and neck squamous cell carcinomas (HNSCC) and their role in disease outcome. In addition, role of herpesviruses 1 and 2 (HSV-1 and -2) and cytomegalovirus (CMV) as co-factors was elucidated. STUDY DESIGN HPV-genotyping of 106 HNSCC was done with Multimetrix(®)-kit. Luminex-based-method was used to detect HSV-1 and -2 and CMV. RESULTS In males, 50% of HNSCC were HPV DNA positive and 25% of these were multiple HPV-types infections and in women, 72% and 31%, respectively. Low-risk (LR) HPV-types were found in 20.5% and co-infection with HSV-1 in 6.6%. Patients with HPV-positive and -negative HNSCC had similar survival. Patients not treated with chemoradiotherapy and co-infected with HSV-1 and HPV had a worse outcome. Similarly patients with LR-HPVs treated with radiotherapy had a poor prognosis. DISCUSSION Radiotherapy for HNSCC in patients with either the presence of LR-HPV-types or a co-infection with HPV and HSV-1 may result in poor outcome.

Vitamin D deficiency predisposes to adherent-invasive Escherichia coli-induced barrier dysfunction and experimental colonic injury.

Background:Adherent-invasive Escherichia coli (AIEC) colonization has been strongly implicated in the pathogenesis of Crohns disease. Environmental triggers such as vitamin D deficiency have emerged as key factors in the pathogenesis of inflammatory bowel diseases. The aim of this study was to investigate the effects of 1,25(OH)2D3 on AIEC infection-induced changes in vivo and in vitro. Methods:Barrier function was assessed in polarized epithelial Caco-2-bbe cells grown in medium with or without vitamin D and challenged with AIEC strain LF82. Weaned C57BL/6 mice were fed either a vitamin D–sufficient or –deficient diet for 5 weeks and then infected with AIEC, in the absence and presence of low-dose dextran sodium sulphate. Disease severity was assessed by histological analysis and in vivo intestinal permeability assay. Presence of invasive bacteria was assessed by transmission electron microscopy. Results:Caco-2-bbe cells incubated with 1,25(OH)2D3 were protected against AIEC-induced disruption of transepithelial electrical resistance and tight-junction protein redistribution. Vitamin D–deficient C57BL/6 mice given a course of 2% dextran sodium sulphate exhibited pronounced epithelial barrier dysfunction, were more susceptible to AIEC colonization, and showed exacerbated colonic injury. Transmission electron microscopy of colonic tissue from infected mice demonstrated invasion of AIEC and fecal microbiome analysis revealed shifts in microbial communities. Conclusions:These data show that vitamin D is able to mitigate the deleterious effects of AIEC on the intestinal mucosa, by maintaining intestinal epithelial barrier homeostasis and preserving tight-junction architecture. This study highlights the association between vitamin D status, dysbiosis, and Crohns disease.

Human papillomavirus in oral atrophic lichen planus lesions

OBJECTIVES Oral lichen planus (OLP) is potentially premalignant disorder in which human papillomavirus (HPV) DNA is detected more often than in normal oral mucosa. We assessed HPV-genotype distribution in atrophic OLPs as related to DNA content and repair, proliferation activity, apoptosis, cell adhesion and lymphocyte infiltration. MATERIALS AND METHODS Eighty-two OLP patients (74.4% women) with a mean follow-up-time of 62.4 months were included in the study. HPV was genotyped with Luminex-based assay detecting 24 HPV-genotypes. Data on a panel of biomarkers and static cytometry performed in these samples before were compared with HPV data. RESULTS HPV DNA was found in 15.9% of OLPs with genotypes: HPV6/11/16/31/33 and one multiple-type infection. Two of the five patients who developed cancer had low-risk HPV6/11 infection while three were HPV-negative. There was a statistically significant correlation between HPV DNA in OLP and DNA content and ploidy markers determined with static cytometry: in HPV-positive samples, proliferation index was higher (p=0.016), less cells were in resting state G1/G0 (p=0.021) but more often in the S-phase (p=0.036) than in HPV-negative lesions. HPV positivity was also related to topoisomerase IIα (p=0.051), caspase-3 (p=0.049) and CD20 (p=0.010) protein expression. CONCLUSION HPV-infection is associated with a subgroup of atrophic OLP. Static cytometry is a sensitive method to identify HPV-associated changes in DNA content and cell proliferation. Of 16 markers, only topoisomerase IIα (proliferation and DNA repair), caspase-3 (apoptosis) and CD20 (B-lymphocytes) were related to HPV. None of the HR-HPVs but only LR-HPVs were associated with the lesions developed to cancer.

Genotype-Specific Incidence and Clearance of Human Papillomavirus in Oral Mucosa of Women: A Six-Year Follow-Up Study

Background There are no previous longitudinal studies on genotype-specific natural history of human papillomavirus (HPV) infections in oral mucosa of women. Methods In the Finnish Family HPV Study, 329 pregnant women were enrolled and followed up. HPV-genotyping of oral scrapings was performed with nested PCR and Multimetrix® test (Progen, Heidelberg, Germany). Incidence and clearance times and rates for each HPV-genotype identified in oral mucosa were determined. Predictors for incident and cleared HPV infections for species 7/9 genotypes were analyzed using Poisson regression model. Results Altogether, 115 baseline HPV-negative women acquired incident oral HPV infection, and 79 women cleared their infection. HPV16 and multiple HPVs most frequently caused incident infections (65% and 12%) in 13.3 and 17.1 months respectively, followed by HPV58, HPV18 and HPV6 (close to 5% each) in 11–24 months. HPV58, HPV18 and HPV66 were the most common to clear. HPV6 and HPV11 had the shortest clearance times, 4.6 months and 2.5 months, and the highest clearance rates, 225.5/1000 wmr and 400/1000 wmr, respectively. The protective factors for incident oral HPV-species 7/9 infections were 1) new pregnancy during follow-up and 2) having the same sexual partner during FU. Increased clearance was related with older age and a history of atopic reactions, whereas previous sexually transmitted disease and new pregnancy were associated with decreased clearance. Conclusions HPV16 was the most frequent genotype to cause an incident oral HPV-infection. Low risk HPV genotypes cleared from oral mucosa more quickly than high risk HPV genotypes. Pregnancy affected the outcome of oral HPV infection.

Lactobacillus rhamnosus GG and its SpaC pilus adhesin modulate inflammatory responsiveness and TLR-related gene expression in the fetal human gut.

Background:Bacterial contact in utero modulates fetal and neonatal immune responses. Maternal probiotic supplementation reduces the risk of immune-mediated disease in the infant. We investigated the immunomodulatory properties of live Lactobacillus rhamnosus GG and its SpaC pilus adhesin in human fetal intestinal models.Methods:Tumor necrosis factor (TNF)-α mRNA expression was measured by qPCR in a human fetal intestinal organ culture model exposed to live L. rhamnosus GG and proinflammatory stimuli. Binding of recombinant SpaC pilus protein to intestinal epithelial cells (IECs) was assessed in human fetal intestinal organ culture and the human fetal intestinal epithelial cell line H4 by immunohistochemistry and immunofluorescence, respectively. TLR-related gene expression in fetal ileal organ culture after exposure to recombinant SpaC was assessed by qPCR.Results:Live L. rhamnosus GG significantly attenuates pathogen-induced TNF-α mRNA expression in the human fetal gut. Recombinant SpaC protein was found to adhere to the fetal gut and to modulate varying levels of TLR-related gene expression.Conclusion:The human fetal gut is responsive to luminal microbes. L. rhamnosus GG significantly attenuates fetal intestinal inflammatory responses to pathogenic bacteria. The L. rhamnosus GG pilus adhesin SpaC binds to immature human IECs and directly modulates IEC innate immune gene expression.

Detection of human papillomavirus in laryngeal squamous cell carcinoma: systematic review and meta-analysis

Recent studies have reported a human papillomavirus (HPV) prevalence of 20% to 30% in laryngeal squamous cell carcinoma (LSCC), although clinical data on HPV involvement remain largely inconsistent, ascribed by some to differences in HPV detection methods or in geographic origin of the studies.

Oral microbiome composition changes in mouse models of colitis.

Oral mucosal pathologies are frequent in inflammatory bowel disease (IBD). Since host–microbiome interactions are implicated in the pathogenesis of IBD, in this study the potential for changes affecting the oral microbiome was evaluated using two complementary mouse models of colitis: either chemically (dextran sulfate sodium) or with Citrobacter rodentium infection.

CD44v6 in developing, dysplastic and malignant oral epithelia

The CD44v6 adhesion molecule has been linked to progression of various carcinomas, but its role in relation to oral-cancer development is not clear. The study was designed to determine whether CD44v6 levels were clinically significant in oral dysplasias. Twenty-nine oral dysplasias were immunostained with CD44v6 antibody on follow-up. Developing normal epithelia and adult normal epithelia and oral carcinomas were stained for comparison. Oral dysplasias and carcinomas exhibited heterogenous staining patterns. No statistically significant correlation between CD44v6 expression and outcome was found for dysplasia patients. The results show that in developing and healthy oral mucosa CD44v6 is associated with epithelium-specific differentiation but in dysplasias and carcinomas it mirrors disorderly epithelial maturation. The results also suggest that determination of CD44v6 levels is not helpful in judging the likely clinical behaviour of oral dysplasia.