Vol 61, No 3 (2016)

The complexity of the pathogenesis and insufficient knowledge about the slow retroviral infections, which include equine infectious anemia, necessitates finding an adequate laboratory model for the study of the infection process and immunogenesis to create means of prevention and treatment of diseases. Data about strains and cellular tropism of the virus are discussed. It was shown that mouse embryonic stem cells (ESCS) exhibited unique properties and characteristics. In contrast to fibroblasts and other cell types, these cells can be considered as a new cell system for studying EIAV in vitro and in vivo. Under differentiation-inducing conditions they are able to reproduce in vitro embryogenesis cells and form cells of three germ layers. Differentiation of mouse ESCs in the direction of hematopoiesis could contribute new knowledge and understanding of viral tropism EIAV in vitro. ESC can be returned back to the early pre-implantation embryo. Once in the germ cell environment, they participate in the formation of tissues and organs of the developing fetus. Thus, the adaptation of the mouse ESC to the equine EIAV through genetic transformation makes it possible to get closer to the creation of a laboratory model for the study of the in vivo immune response in the lentiviral infection.

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.

In this work, the results of a comprehensive laboratory examination of 37 children with retinoblastoma were described. The presence of Igm-, IgA, - IgG- antibodies to the herpes simplex virus types 1 and 2, cytomegalovirus (СMV), epstein-Barr virus (eBV), human herpes virus (HHV) type 6, Toxoplasma gondii, mycoplasma hominis and ureaplasma urealyticum in the serum was tested using ELISA. In the polymerase chain reaction the DNA of these pathogens were detected in the blood plasma of 18 patients and tumor biopsy specimens from 10 eyes. The results showed that children with RB were predominantly infected by the herpesviruses, among which prevailed CMV. in 4 of 5 enucleated eyes the DNA of herpesvirus [CMV (2 eyes), EBV (1 eye), HHV 6 (1 eye)] and ureaplasma urealyticum (1 eye) were also present in tumor tissue. Nucleic acid of infectious microorganisms were considerably more often detected in the tumor tissue than in plasma (5 of 10, 1 of 18, respectively; p = 0.023), suggesting thereby the presence of the virus in the eye and its adverse role in the pathogenesis of the RB.

A possible approach to effective, pathogenetically valid treatment of patients with the tick-borne encephalitis (TBE) is a complex therapy with the immunotropic preparations isolated from natural objects. This work is devoted to the comparative study of the antiviral activity of the tinrostim (immunoactive peptide from the optical ganglia of the squid Berritiuthis magister) and some officinal drugs used for prevention and treatment of the TBE (ribavirin, reaferon-EC, cycloferon, 4-jodantipyrin, immunoglobulin human against encephalitis ixodicum) in the experimental models of the TBE. All tested drugs significantly inhibited the proliferation of the highly virulent strain of the TBEV in the sensitive PK cell cultures: ribavirin and immunoglobulin against TBE completely inhibited viral replication (by 100%); cycloferon – by 75%; tinrostim, reaferon-EC, and jodantipyrin – by 50-60%. Therapeutic efficacy of the compounds was evaluated on a model of acute lethal TBE in mice: treatment with cycloferon and immunoglobulin against TBE prevented the mortality in 35-45% of infected animals; tinrostim – in 25%; ribavirin, reaferon-EC, and jodantipyrin – in 5-10%. The combination of the immunoactive peptide, tinrostim, with officinal drugs (ribavirin, cycloferon) was more effective than the treatment with a single drug, thereby indicating the prospects of the use of this therapy for treating TBE.