Goal of the study. To study the dependence of the clinical course of the papilloma viral infection and cytological characteristics of lesions in the cervical mucosa on the quantitative indices of human papilloma virus (HPV). Materials and methods. The study involved 175 female patients with HPV of a high carcinogenic risk including 125 subjects with clinical forms of the papilloma viral infection (PVI) and 50 subjects with subclinical and latent forms of the disease. Laboratory tests were carried out with the use of the polymerase chain reaction including real-time PCR for the quantitative determination of HPV. Cytological examinations of scrapes from the exocervical and endocervical mucosa were carried out according to Leishman I, and the results were interpreted according to Bethesda. Results. The authors established an association between clinical forms of PVI and infection with two or more HPV genotypes, and latent and subclinical forms of the disease and infection with one HPV genotype; HPV Genotype 16 prevails within the structure of HPV of a high carcinogenic risk. It was shown that patients infected with two or more HPV genotypes as well as subjects with subclinical and latent forms of the disease underwent reliably higher viral loads than subjects with anogenital warts and patients infected with one HPV genotype only. A higher HPV viral load was also noted in case of a persisting course of PVI and in patients with high-grade squamous intraepithelial lesions (H-SIL). Conclusion. Women with latent and subclinical forms, persistent PVI course and infected with two or more HPV genotypes of a high carcinogenic risk belong to the high-risk group developing expressed epithelial affections in the cervical mucosa. Quantitative HPV indices exceeding 5 lg of copies of HPV DNA per 100,000 cells belong to unfavorable predictors for the development of intraepithelial affections in the cervical mucosa and stipulate the need to conduct an additional examination (colposcopy or cytology) to exclude their development.

Aim: To study an association between seborrheic keratosis and human papilloma virus (HPV) using quantitative analysis of viral desoxyribonucleic acid (DNA); to assess prevalence of different phenotypes of beta-HPV. Materials and methods: We examined 60 renal transplant recipients (20 of them had multiple seborrheic keratosis) and 22 immunocompetent patients with seborrheic keratosis. Control group included 49 healthy subjects. Burr biopsy samples (micro-samples) were collected in sterile conditions. After sample procession and DNA isolation using DNK-sorb-C kit (Central Research Institute for Epidemiology – CRIE), polymerase chain reaction for HPV was performed with real-time fluorescent hybridization detection. For DNA amplification and detection we used RotorGene 3000 analyzer (Corbett Research, Australia). In the beta-HPV assay, recombinant plasmids were used as positive controls and control human beta-globin gene fragments (CRIE). 4 oligo-nucleotide systems (group-specific primers and probes) were used for the detection of beta-HPV DNA. Results: Keratotic lesions of open and covered skin regions were common in renal transplant recipients. Beta-HPV DNA was more frequent in seborrheis keratomas and intact skin (81% and 55%) of renal transplant recipients compared to healthy donors (47%). Conclusion: HPV DNA was frequently detected in keratotic lesions and intact skin of renal transplant recipients. In immunocompetent patients prevalence of HPV DNA in keratotic lesions was significantly higher compared to intact skin.

A battery of tests for detection human papillomavirus DNA, mRNA corresponding to viral oncogenes, and viral oncoprotein E7 in cancer bladder urothelium was piloted in 35 samples of bladder cancer. DNA of human papillomavirus type 16 (causes cervical cancer) was found in 16 (46%) samples; E6/E7 oncogene transcript and E7 oncoprotein of human papillomavirus type 16 were detected in 10 and 7 human papillomavirus DNA-positive samples, respectively. These findings attest to association of bladder cancer with human papillomavirus in Russia.

The DNA of virus of human papilloma of high carcinogenic risk was detected in 116 cervical samples. At that, the morphological symptoms of background processes are detected in 19 samples, CIN 1 in 9, CIN 2 in 23, CIN 3 in 54 (and out of them carcinoma in situ in 13), epidermoid cancer (squamous cell carcinoma) in 11 cases. The viral load of human papilloma of high carcinogenic risk in all samples of DNA exceeded threshold of clinical value (3 lg copies of DNA of human papilloma/105 cells). The genetic typing of human papilloma of high carcinogenic risk revealed the dominance of human papilloma of type 16 in 49.7%, type 33 in 15.3%, type 31 in 12.3% and type 45 in 5.5%. In women with background processes in cervix of the uterus DNA of human papilloma type 16 was detected more often in episome form. In case of dysplastic alterations of epithelium and cervical cancer DNA of human papilloma type 16 is detected in mixt form with different degree of integration into cell genome.

The DNA of virus of human papilloma of high carcinogenic risk was detected in 116 cervical samples. At that, the morphological symptoms of background processes are detected in 19 samples, CIN 1 in 9, CIN 2 in 23, CIN 3 in 54 (and out of them carcinoma in situ in 13), epidermoid cancer (squamous cell carcinoma) in 11 cases. The viral load of human papilloma of high carcinogenic risk in all samples of DNA exceeded threshold of clinical value (3 lg copies of DNA of human papilloma/105 cells). The genetic typing of human papilloma of high carcinogenic risk revealed the dominance of human papilloma of type 16 in 49.7%, type 33 in 15.3%, type 31 in 12.3% and type 45 in 5.5%. In women with background processes in cervix of the uterus DNA of human papilloma type 16 was detected more often in episome form. In case of dysplastic alterations of epithelium and cervical cancer DNA of human papilloma type 16 is detected in mixt form with different degree of integration into cell genome.

A female patient with recurrent bladder cancer underwent complex examination. The primary tumor removed in 2004 showed human papillomavirus (HPV) 16 DNA, mRNA corresponding to HPV16 oncogene E7, as well as HPV16 protein E7. The patient is a smoker who has been working at a chemical factory for over 20 years. During tumor recurrence in 2009, there was no DNA of high-risk HPV types in the cancer cells. HPV16 E7protein and cellular p 16(INK4alpha), an indicator of HPV-induced carcinogenesis, were not found. Colposcopy revealed no precancerous changes in the epithelium of the cervix uteri. The cervical epitheliocytes contained no high-risk HPV DNA, E7 and p16(INK4alpha) proteins. It seems expedient to continue in vitro studies of the possible role of HPV in urothelial carcinogenesis on an experimental model.

Резолюция по итогам Совета экспертов по иммунотерапии распространенного рака мочевого пузыря

Hyperexpression of p16(INK4a) protein is an early marker of cervical cancer. Hyperexpression of INK4a gene encoding this protein at the level of mRNA and p16(INK4a) was detected in tumor cells of some patients with bladder cancer associated with human papilloma virus-16. However, in contrast to cervical cancer, this phenomenon in urothelial carcinomas does not correlate with expression of human papilloma virus-16 oncogenes E6 and E7.

Oncoprotein E7 HPV16 was detected by immunohistochemical staining with specific polyclonal antiserum [Fiedler et al., 2004] in 7 out of the 24 (29.2%) studied bladder cancer specimens. The result is in good agreement with the hypothesis that HPVs take part in the carcinogenesis of the urothelium. However, some of the observations made seem rather hard to be interpreted at present. The latter include the detection of E7 HPV16 in a small number of cancer cells in a few bladder cancer specimens being examined; the presence of this protein in the cytoplasm, rather in the cancer cell nuclei, and its detection in some morphologically normal bladder urothelial specimens from non-cancer patients. Thus, the hypothesis that HPVs are implicated in the carcinogenesis of the bladder urothelium deserves further verification.