Detection of human cytomegalovirus in motile spermatozoa and spermatogenic cells in testis organotypic culture

Herpesviridae

Alla A Kushch¹  Sergei V Alkhovskii¹  Anton S Tsibizov¹  Regina R Klimova¹  Larisa E Zavalishina⁴  Aleksander S Segal⁵  Ludmila V Shileyko²  Lubov' F Kurilo²  Sergei A Yakovenko³  Yurii A Tyulenev¹  Victor A Naumenko¹ 

[1] The D. I. Ivanovsky Institute of Virology, Ministry of Health and Social Development of the Russian Federation, 123098 Gamaleya str. 16, Moscow, Russia;Medical & Genetic Research Center of Russian Academy of Medical Sciences, 115478 Moscworechye str. 1, Moscow, Russia;Altravita IVF Clinic, 117186 Nagornaya str. 4a, Moscow, Russia;P.A. Herzen Research Oncological Institute, 125284 2nd Botkinsky pr. 3, Moscow, Russia;Department of Urology, Moscow State Medical Dental University, 127473 Delegatskaya str. 20/1, Moscow, Russia

关键词: testis organotypic culture;    spermatogenesis;    infertility;    human cytomegalovirus;   

Others  :  801370
DOI  :  10.1186/2042-4280-2-7

 received in 2011-04-11, accepted in 2011-06-28,  发布年份 2011

【 摘 要 】

Background

The presence of human cytomegalovirus (HCMV) in male genital tract suggests its vertical transmission with spermatozoa and the development of a potentially dangerous fetal infection. The objective of the present study was to evaluate the possibility of intracellular HCMV localization in male germ cells and to examine the effect of the virus on human spermatogenesis.

Methods

Semen samples from 91 infertile and 47 fertile men were analyzed. HCMV was detected by real time PCR, rapid culture method and PCR in situ. Human testis organotypic culture and quantitative karyological analysis were used to investigate viral effects on spermatogenesis. Localization of HCMV in immature germ cells and spermatozoa was studied by immunostaining with monoclonal antibodies and ultrastructural analysis of infected organotypic culture.

Results

Viral DNA was detected in 12.3% samples of motile spermatozoa, while infectious activity only in 2.9% infertile and fertile men without statistically significant intergroup difference. According to PCR in situ, the mean percentage of infected cell in both groups was 1.5% (0.25%-15%), which can serve as a criterion for evaluating the risk of HCMV transmission. In HCMV-infected organotypic culture viral antigens were identified in spermatides on day 4, in spermatogonia and spermatocytes on day 8, and in spermatozoa on day 14. Empty and full capsides and virions were visualized in germ cells by electron microscopy. The number of cells before introduction in culture was taken for 100%. On day 14 infected culture contained 36.8% spermatogonia, 18.7% spermatocytes, 27.6% round spermatides and 42.5% elongated spermatides; in comparison with 82.2%, 51.5%, 70.4% and 65.7% in uninfected culture, respectively (all p < 0.05). There were no changes in the number and viability of spermatozoa.

Conclusions

HCMV was detected in male germ cells, both in sperm samples and in testis organotypic culture. The virus may infect immature germ cells which develop to mature HCMV-carrying spermatozoa. A considerable decrease in the number of immature germ cells indicates that HCMV produces a direct gametotoxic effect and can contribute to male infertility.

【 授权许可】

   
2011 Naumenko et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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