Projects / Programmes
HPV infection in Slovene fertile and infertile men, its harmful effect on sperm and transmission to female partners: arguments for early diagnosis and prevention
| Code |
Science |
Field |
Subfield |
| 3.05.00 |
Medical sciences |
Human reproduction |
|
| Code |
Science |
Field |
| B570 |
Biomedical sciences |
Obstetrics, gynaecology, andrology, reproduction, sexuality |
| Code |
Science |
Field |
| 3.02 |
Medical and Health Sciences |
Clinical medicine |
infertile men, human paipllomavirus infection, sperm quality, apoptosis, mitochondrial membrane potential, DNA fragmentation, electron microscopy, testicular histology, prevalence
Researchers (24)
Organisations (3)
Abstract
Etiologically, human papillomaviruses (HPV) are associated with benign and malignant lesions in cells of skin and mucous epithelium in both sexes, but little is known about the effect of HPV on fertility, especially in males. The presence of HPV DNA in sperm is most frequently associated with decreased sperm motility.
The aim of this study is to determine the effect of HPV infection on male fertility. We aim at finding the association between the presence of HPV DNA in the collected samples of the couple and sperm quality. Our focus will be on genotype-specific prevalence of HPV DNA infection in men of infertile couples. We will determine which organ, testis, epididymis or prostate malfunctions at the presence of HPV DNA. We will further search for the mechanism of harmful effects of HPV on the sperm cell. The women, partners of men affected by HPV infection will be followed up by HPV DNA determination and cytologic smear taking.
The study will be carried out at the Department of Obstetrics and Gynecology in collaboration with the Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana and the Center for Animal Genomics. Veterinary Faculty, University of Ljubljana. We will enrol 50 infertile and 50 fertile couples. To determine the prevalence of HPV DNA, additional 250 men of infertile couples will be enrolled. The couple will be taken five samples: cervical smear and a urine sample from the female partner, and a scrape from the surface skin of the penis, urine and a semen sample. For determination of HPV DNA prevalence in infertile men, the 3 lastly mentioned samples will be used.
All the samples with the exception of semen sample, will be immediately prepared and stored at -20°C until DNA extraction. The sperm sample will be managed at the andrology laboratory of the Department: one part will be submitted to HPV DNA extraction, while the other part will be prepared by centrifugation in the density gradient for the analysis of classic sperm characteristics and determination of sperm apoptotic markers (membrane mitochondrial potential (MMP) and sperm DNA fragmentation).
Sperm apoptotic markers will only be identified in sperm samples of men of fertile and infertile couples. The measurement of sperm apoptotic markers is performed using flow cytometry. MMP will be measured using DiOC6(3) staining, DNA sperm fragmentation will be determined using the TUNEL method.
DNA extraction from all frozen samples will be carried out using QIAampâ DNA Mini Kit (QIAGEN, Gaithersburg, USA) kit. Genotypes will be determined using LINEAR ARRAY HPV Genotyping test (Roche, Branchburg, NJ, USA).
Additionally, alpha glucosidase (epididymal function) and citron acid (prostatic function) will be determined. In men with HPV infection electron microscopy of sperm from the ejaculate and testis will be performed. From bioptic samples of testicular tissue of azoospermics HPV DNA will be extracted, and correlations with histology searched for. In women, partners of men with HPV infection, cervical smear for cytologic examination and HPV DNA isolation will be taken.
The results will determine the association between HPV and fertility. HPV DNA is more frequently found in semen of infertile men compared with fertile men. In men with HPV DNA in semen, classic characteristics of sperm and/or sperm apoptotic markers are abnormal. In azoospermics with HPV DNA in testicular tissue abnormal spermatogenesis is more frequent, and epididymal and prostatic functions are also abnormal. The analysis of morphology with electron microscopy of sperm from the ejaculate or testis will reveal the mechanisms of DNA invading the sperm cell. In female partners of men with HPV infection, HPV or cervical dysplasia occurs within two years since the screening of her male partner. Asymptomatic HPV infection in sperm is related with male infertility and de novo occurrence of HPV DNA cervical cells, which is why vaccination of men with HPV is
Significance for science
In representative populations of males from infertile couples vs. control group, we demonstrated that no correlations exist between HPV DNA in semen and decreased sperm motility, concentration and morphology considering the WHO criteria. Also, the correlation between HPV DNA in semen and pathological results of sperm function tests was not found. The FISH technique revealed that the most frequent site of HPV DNA was the surface of sperm head.
Significance for the country
Slovenia is today one of the few countries in the world which tested for HPV and reported the high HPV DNA prevalence in infertile and fertile males. Up today the prevalence of HPV was documented only in women. As regard to two papers published this year by our group, Slovenia would lead additional research in order to clear the relationship between HPV and assisted reproductive technologies.
Most important scientific results
Annual report
2011,
2012,
2013,
final report,
complete report on dLib.si
Most important socioeconomically and culturally relevant results
Annual report
2011,
2013,
final report,
complete report on dLib.si
