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

Abstract

BackgroundThe 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.MethodsSemen 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.ResultsViral 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.ConclusionsHCMV 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.

Detection and quantification of human herpes viruses types 4–6 in sperm samples of patients with fertility disorders and chronic inflammatory urogenital tract diseases

“…In some infected cells, we found an equatorial pattern of staining similar to the attachment site described for human papillomavirus (Foresta et al ., ). It has previously been shown that CMV can infect spermatogonia, spermatocytes and spermatides in human testis organotypic culture (Naumenko et al ., ). To our knowledge, this is the first report of successful CMV infection of spermatozoa in vitro.…”

Describing the Silent Human Virome with an Emphasis on Giant Viruses

“…Asymptomatic shedding from the genito-urinary tract was reported mainly for dsDNA eukaryotic viruses of the Adenoviridae, Herpesviridae, Papillomaviridae and Polyomaviridae families with the exception of ssDNA viruses of the Anelloviridae family [83,106,107,108,109,110,111]. In the case of polyomaviruses, it appears that viral excretion was correlated with the host immune status.…”

“…In this case it appears that some herpesviruses, such as human Herpesvirus 6 A/B or the Cytomegalovirus, were able to attach to the sperm head with an intact acrosome [108,113]. Thus, given the potential risk some herpesviruses may represent to the newborns, additional research is required to evaluate the impact of this asymptomatic shedding from herpesvirus-positive donor semen.…”

Herpes simplex virus inoculation in murine rete testis results in irreversible testicular damage

“…HSV and another member of the Herpesviridae, human cytomegalovirus (CMV), were found to replicate and cause damage to somatic and germ cells in testis organotypic culture (Naumenko et al . ,b). Despite the significance of these observations, it remains unclear whether HSV can cause testicular infection in vivo .…”