Outcomes of HPV type-specific serostatus do not associate with oral or genital HPV-carriage in non-vaccinated women followed for three years - BMC Women's Health - bmcwomenshealth.biomedcentral.com

Subjects

The Finnish Family HPV (FFHPV) study is a prospective cohort study conducted at the Department of Obstetrics and Gynecology, Turku University Hospital, University of Turku and at the Institute of Dentistry, Faculty of Medicine, University of Turku, Turku, Finland. The cohort included 329 pregnant women who were recruited between 1998 and 2001 (minimum of 36 weeks of pregnancy) and were followed-up for six years after the delivery. All participants were Caucasian origin and had the same ethnic background. The HPV results of the whole FFHPV study cohort (331 mothers and 131 fathers) have been reported in a series of previous publications, only few cited here [1, 12, 13]. A written informed consent was obtained from all the study participants. The Research Ethics Committee of Turku University and Turku University Hospital has approved the study protocol and its amendment (#2/1998 and #2/2006).

The present study focuses on naturally acquired HPV antibody levels for HPV genotypes 6, 11, 16, 18 and 45 and their association to the genital and oral HPV infection outcomes among 267 unvaccinated women from the FFHPV study [14]. Of the 329 women originally enrolled, 267 women were eligible for the present study having at least two serum samples taken during the study period.

Samples

Genital and oral scrapings from the women were collected for HPV-testing with a cytobrush (MedScand, Malmö, Sweden) as described before [12]. HPV genotyping was done between the years of 2005 to 2010 by Luminex-based Multimetrix kit (Progen Biotechnik GmbH, Heidelberg, Germany), which detects 24 low-risk (LR)- and high-risk (HR)-HPV genotypes as followed: LR-HPV: 6, 11, 42, 43, 44, and HR-HPV: 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, 82 [15].

Serology

Blood samples were taken at baseline and at 12-, 24- and 36 months of the follow-up. The samples were collected between the years of 1998 to 2003. All samples were stored after collection first at − 20 °C for no longer than one week, and then at − 70 °C until the analyzed between the year 2008 to 2009 as previously described [1]. Major capsid protein L1 antibodies for HPV types 6, 11, 16, 18 and 45 were analyzed by multiplex HPV serology based on glutathione S-transferase fusion-protein capture on fluorescent beads (also referred as GST-L1 assay), as described previously [16, 17]. This method is frequently used and validated in seroepidemiological studies concerning HPV [18]. Sera were scored as positive when the antigen-specific median fluorescence intensity (MFI) values exceeded the cut-off level of 200 MFI for the L1 antigen of individual HPV genotypes [19].

Statistical analyses

Frequency tables were analyzed using the χ2 test or the Fisher's exact test for categorical variables. Differences in the means of continuous variables (i.e. log-transformed HPV antibody titres) were analyzed using ANOVA (analysis of variance) after controlling for their normal distribution [14]. Women's genital and oral HPV 6, 11, 16, 18 and 45 genotype-specific and any-HPV prevalence at each follow-up visit was compared with the women's serological status for these same HPV genotypes [14]. Serological status was classified into three categories as follows: 1) Always seronegative (MFI remains < 200 at each visit); 2) Seroconversion, defined by two conditions: (i) an MFI value < 200 in the first and > 200 in the subsequent sample, and (ii) at least a two-fold increase of the previous serum value in any subsequent sample; and lastly 3) Persistent seropositivity (MFI constantly > 200 in all follow-up visits) [14]. The persistence and clearance of oral and genital HPV 6, 11, 16, 18 and 45 infections were compared between two groups of women: 1) those with constantly high-titer (> 400MFI) of HPV antibodies, and 2) women who tested constantly HPV-seronegative. Logistic regression with its Odd Ratio (OR) was calculated as a likelihood of the above serostatus to predict HPV persistence or clearance. Persistent HPV infection was defined as being positive for type-specific 6,11,16,18 or 45 HPV genotype for 24 months or longer during the follow-up. All statistical analyses were performed using SPSS (IBM, NY, USA, PASW Statistics version 26.0.1.) and STATA/SE 16.1 (Stata Corp., College Station, TX, USA) software packages. All statistical tests performed were two-sided and declared significant at the P-value < 0.05 level.

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