Interaction of herpesviruses with mature human spermatozoa in the model system  in vitro

E. A. Malolina; Малолина Е. А.; A. L. Lebedeva; Лебедева А. Л.; A. Yu. Kulibin; Кулибин А. Ю.; V. V. Evdokimov; Евдокимов В. В.; L. F. Kurilo; Курило Л. Ф.; T. M. Sorokina; Сорокина Т. М.; Ju. A. Tulenev; Тюленев Ю. А.; V. A. Naumenko; Науменко В. А.; A. A. Kushch; Кущ А. А.

doi:10.18821/0507-4088-2016-61-3-119-125

Interaction of herpesviruses with mature human spermatozoa in the model system in vitro

Authors: Malolina E.A.¹^,2, Lebedeva A.L.¹^,3, Kulibin A.Y.², Evdokimov V.V.¹^,4, Kurilo L.F.³, Sorokina T.M.³, Tulenev J.A.¹, Naumenko V.A.¹, Kushch A.A.¹
Affiliations:
1. Federal Research Centre of Epidemilogy and Microbiology named after the honorary academician N.F. Gamaleya
2. Koltzov Institute of Developmental Biology
3. Research Center for Medical Genetics
4. Moscow Scientific and Practical Center of Dermatovenereology and Cosmetology, Department of Healthcare of the Government of Moscow
Issue: Vol 61, No 3 (2016)
Pages: 119-125
Section: ORIGINAL RESEARCH
URL: https://virusjour.crie.ru/jour/article/view/370
DOI: https://doi.org/10.18821/0507-4088-2016-61-3-119-125
ID: 370

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Abstract

The DNA of human herpesviruses (HHV), including the herpes simplex virus (HSV) and cytomegalovirus (CMV), is often identified in ejaculates of patients with urogenital diseases and infertility. At least a part of viral DNA is associated with cell fraction of ejaculate. However, it remains unclear how the semen is infected by the virus. It can be located in gametes or be capable of infecting mature germ cells, including motile sperm cells. In order to resolve this issue, interactions of the CMV and HSV with human sperm cells were studied using an original optimized model of the herpesviral infection of male gametes in vitro. The analysis of the immunofluorescent staining of gametes for viral antigens has shown that CMV infected 2% gametes, while HSV infected 17.26 ± 2.58% gametes. The fraction of progressively motile sperm cells contained 13.99 ± 4.64% infected cells. Localization of HSV was studied by the confocal microscopy. Sometimes, viral gB protein was found on sperm cell membrane. In addition, optical scanning of other cells has shown the intracellular localization of the viral proteins. In the majority of spermatozoa, the viral proteins were observed in the head and neck. In some cells, they were located in the middle piece or, rarely, in the equatorial segment. In general, after in vitro infection HSV antigens were located in the same areas of the sperm cells as in ejaculates from infected patients. According to DNA–DNA hybridization in situ, gametes containing HSV DNA accounted for 16.94 ± 5.28%, which is consistent with the results obtained in the immunofluorescence assay. It can be concluded that mature male gametes are infected by HHV in the genital tract, where the virus binds to the sperm cell membrane and enters the cell. Interaction of HHV with progressively motile sperm cells implies a vertical viral transmission upon fertilization and points to the necessity of testing ejaculate for herpesviruses infections.

Keywords

human herpesviruses, herpes simplex virus, cytomegalovirus, human sperm, infection of gametes in vitro, male infertility, vertical transmission, DNA-DNA hybridization, localization of viral antigens, confocal microscopy

About the authors

E. A. Malolina

Federal Research Centre of Epidemilogy and Microbiology named after the honorary academician N.F. Gamaleya; Koltzov Institute of Developmental Biology

Author for correspondence.
Email: kate.ma85@gmail.com
ORCID iD: 0000-0003-3539-6829

Ekaterina A. Malolina, Candidate of Biology, Research scientist

Moscow, 123098
Moscow, 119334

Russian Federation

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