[Antiviral activity of the organic germanium complex with aciclovir against herpes simplex virus (Herpesviridae: Alphaherpesvirinae: Simplexvirus: Human alphaherpesvirus 1/2) in the in vitro and in vivo systems].

INTRODUCTION: A significant increase in the incidence of various forms of herpesvirus infection (HVI) determines the need to search for new approaches to the modification of one of the basic antiviral drugs aciclovir (ACV) and its dosage forms to improve their biopharmaceutical characteristics and increase the effectiveness of therapy. In this aspect, an innovative organic germanium complex with aciclovir (OGCA) is promising.The aim of the study was to assess the antiviral activity of OGCA against the herpes simplex virus (HSV) (human herpes virus, HHV) on the HVI models both in vitro and in vivo. MATERIAL AND METHODS: We studied the activity of OGCA in a therapeutic regimen against HSV-1 (HHV-1) (Kl strain), HSV-2 (HHV-2) (VN strain) using virological and statistical research methods in the in vitro model of HVI on Vero cell culture and the model of genital herpes (GH) caused by HHV-2 (VN strain) in male guinea pigs (Canis porcellus). RESULTS AND DISCUSSION: It was found OGCA inhibits the replication of HHV-1 and HHV-2 in Vero cells, and has anti-HHV activity in the GH model in male guinea pigs, leading to a decrease in the severity and duration of the disease, the intensity and duration of viral shedding. The most pronounced activity was detected when preparation was applied topically 5 times a day for 5 days at the early stages of infection (3% gel). The delayed use of OGCA (48 hours after infection) also had statistically significant efficacy compared to commercial reference drugs containing aciclovir or its pro-drugs: aciclovir (5% cream), AIL (acyclovir+interferon alfa-2b+lidocaine, 3% ointment), penciclovir (1% cream). OGCA significantly reduced the number of days of the pathogen shedding, as well as its infectivity, compared to animals in the control group and ones receiving placebo. The activity of OGCA, apparently, is due to its improved biopharmaceutical characteristics compared to aciclovir, as well as the presence of a number of biological activities of its constituent components. CONCLUSION: The results of the study allow us to consider OGCA as the basis for the development of antiviral agents for the treatment of HVI.

[RETRACTED: High-throughput sequencing in diagnostics and prevention of herpes simplex virus infection (Herpesviridae, Alphaherpesvirinae, Simplexvirus, Human alphaherpesvirus 1)].

RETRACTEDHerpes simplex viruses types 1 (HSV-1) and 2 (HSV-2) are among the most common viruses in the human population. The clinical manifestations of HSV infection vary widely, which necessitates reliable molecular methods for the timely diagnosis of herpes virus infection, as well as for detection of mutations in the genes responsible for drug resistance. PCR is often unable to detect HSV isolates with nucleotide substitutions at the primer binding site. Sanger sequencing of the whole genome reveals mutations mainly at the consensus level, which accumulate at advanced stages of viral infection. High-throughput sequencing (HTS, next generation sequencing) offers an obvious advantage both in early diagnosis of herpes virus infection and identification of HSV variants.

Development of the suppository form of human immunoglobulin preparation with high titers of antibodies to herpes simplex virus types 1 and 2 for the treatment of chronic forms of herpetic disease.

In spite of the vast arsenal of therapeutic agents, therapy of herpes virus infection (HVI) is very difficult, particularly in pregnant women, newborns and children in the first years of life, as well as in patients with immune deficiency. In this regard, possibility of using immunoglobulins for the treatment of HVI is currently attracting the attention of doctors. The aim of this work was to develop a suppository form of the drug containing donor immunoglobulins with high levels of neutralizing antibodies to herpes simplex virus types 1 and 2 for the treatment of chronic forms of herpetic disease. The study included the following steps: 1) selection of gamma-globulins with high antibody titer for HSV-1 and HSV-2 ELISA test; 2) determination of the level of neutralizing antibodies in the selected series of gamma-globulins in tests in tissue cultures and animals; 3) lyophilization of immunoglobulins; 4) development of the suppository form of the preparation containing gamma-globulin donors with high levels of neutralizing antibodies to HSV-1 and HSV-2; 5) study of the safety of the activity of neutralizing antibodies to HSV-1 and HSV-2 in the suppository form of the drug with hyaluronic acid used as immunomodulator. As the result of this work, immunoglobulin preparation in the suppository form was developed. The developed preparation meets the requirements for safety and efficacy. It is not toxic or pyrogenic. The problems of clinical use of this drug as a method of HVI therapy are discussed.

[THE SPECTRUM OF MARKERS OF HERPES VIRAL INFECTIONS AND ALGORITHM OF THEIR LABORATORY DIAGNOSTIC IN CHILDREN WITH INFLAMMATORY PROCESSES OF UPPER RESPIRATORY WAYS AND ENT-ORGANS].

The purpose of study is to explore markers of persistent herpes viral infections in children with inflammatory processes of upper respiratory ways and ENT-organs. The sampling included 118 examined patients aged from 1 month to 17 years. The complex of standardized viral, immunological, molecular genetic methods was applied to detect (to exclude) herpes infection: cytomegalovirus infection, Epstein-Barre virus infection, simplex herpes virus infection. The diagnostic algorithm of examination of children with diseases of upper respiratory ways for herpes infection is presented. The dominating significance of simplex herpes virus and Epstein-Barre virus and also Staphylococcus aureus and Streptococcus haemolyticus-ß group A at the analysis of microbial landscape. In 83.9% of children with diseases of upper respiratory ways chronic infections of simplex herpes virus, Epstein-Barre virus, cytomegalovirus; in39.39% - mixed-infection; in 41.03% - combination of simplex herpes virus and Epstein-Barre virus infections; in 33.33% - combination of simplex herpes virus and cytomegalovirus infections; in 7.69% - combination of simplex herpes virus and Epstein-Barre virus and cytomegalovirus infections; in 17.94% - combination of Epstein-Barre virus and cytomegalovirus infections; The particularity of course of persistent herpes infection in children had to do with absence of specific symptoms of nosologic form in 59.2% of cases. The results of analysis of smears from nasopharynx of children infected with herpes viruses permitted to detect: Staphylococcus aureus in 36.36%; Streptococcus haemolyticus-ß in 32.32%; Streptococcus haemolyticus-α in 11.11%; Candida albicans of mucous membranes in 4.04% of children. The viral bacterial mixed-infection was detected in 44.44%. The laboratory signs of activity of immune inflammation were detected: increasing of content of TNАα and decreasing of level of IFNγ. The results of study substantiate necessity of individual approach to therapy of children with diseases of upper respiratory ways and ENT-organs and with implementation of complex of curative rehabilitating activities.The purpose of study is to explore markers of persistent herpes viral infections in children with inflammatory processes of upper respiratory ways and ENT-organs. The sampling included 118 examined patients aged from 1 month to 17 years. The complex of standardized viral, immunological, molecular genetic methods was applied to detect (to exclude) herpes infection: cytomegalovirus infection, Epstein-Barre virus infection, simplex herpes virus infection. The diagnostic algorithm of examination of children with diseases of upper respiratory ways for herpes infection is presented. The dominating significance of simplex herpes virus and Epstein-Barre virus and also Staphylococcus aureus and Streptococcus haemolyticus-ß group A at the analysis of microbial landscape. In 83.9% of children with diseases of upper respiratory ways chronic infections of simplex herpes virus, Epstein-Barre virus, cytomegalovirus; in39.39% - mixed-infection; in 41.03% - combination of simplex herpes virus and Epstein-Barre virus infections; in 33.33% - combination of simplex herpes virus and cytomegalovirus infections; in 7.69% - combination of simplex herpes virus and Epstein-Barre virus and cytomegalovirus infections; in 17.94% - combination of Epstein-Barre virus and cytomegalovirus infections; The particularity of course of persistent herpes infection in children had to do with absence of specific symptoms of nosologic form in 59.2% of cases. The results of analysis of smears from nasopharynx of children infected with herpes viruses permitted to detect: Staphylococcus aureus in 36.36%; Streptococcus haemolyticus-ß in 32.32%; Streptococcus haemolyticus-α in 11.11%; Candida albicans of mucous membranes in 4.04% of children. The viral bacterial mixed-infection was detected in 44.44%. The laboratory signs of activity of immune inflammation were detected: increasing of content of TNАα and decreasing of level of IFNγ. The results of study substantiate necessity of individual approach to therapy of children with diseases of upper respiratory ways and ENT-organs and with implementation of complex of curative rehabilitating activities.

[SYNTHETIC PEPTIDE VACCINES].

An update on the development and trials of synthetic peptide vaccines is reviewed. The review considers the successful examples of specific protection as a result of immunization with synthetic peptides using various protocols. The importance of conformation for the immunogenicity of the peptide is pointed out. An alternative strategy of the protection of the organism against the infection using synthetic peptides is suggested.

[Molecular genetic characteristics of the virus isolated from patients with human acute encephalomyelitis and multiple sclerosis].

The study of the antigenic and molecular genetic structure of human acute encephalomyelitis virus (HAEV) showed a high similarity of the HAEV N gene with the homologous gene of the fixed rabies virus strain. The results of the nucleotide sequence analysis indicate that HAEV belongs to the lyssavirus genotype 1. The N gene sequence is the closest to those of the ERA-CB20-M and RV-97 strains of the rabies virus. The need for further research into the role of the human acute encephalomyelitis virus in human pathology stems from past surveys that revealed the presence of the VNAs against this virus in 6 per cent of the blood received from donors in the USA and in each third among the patients with multiple sclerosis in the former USSR.

Population structure of some street rabies virus strains.

Street rabies virus strains can contain from one to three biological (clinical) variants as it has been estimated using random-bred white mice and dogs.

Effect of adenovirus infection on human peripheral lymphocytes.

The effect of adenovirus infection on human peripheral lymphocytes has been studied in vitro. Virus antigens were found to multiply in the cell cultures but no infective virus was formed. On infecting the lymphocytes after stimulation with phytohaemagglutinin, infective virus was formed though in low titre. Adenovirus infection decreased the E-rosette forming activity and the response to phytohaemagglutinin of the lymphocytes. The immunomodulating drug levamisole failed to influence significantly the effect of the virus. Adenovirus thus causes several changes in the characteristics of peripheral lymphocytes which may lead to a disturbance of the immune function.

Development of tumours in mice chronically infected with herpes simplex virus.

Tumours developed in chronic infection lasting for 150--180 days in 39 (60%) of 65 mice infected with strain L2 of herpes simplex virus (HSV) type 1 and in 12 (20%) of 60 mice infected with strain 333 of HSV type 2. Similar results were obtained in 150 immunosuppressed mice chronically infected with HSV types 1 and 2. Pathomorphologically, the neoplasias in the first group (strain L2) were similar to adenocarcinoma and malignant lymphoma and in the second (strain 333) to lymphoma and angio- or fibrosarcoma. The respective HSV strains were isolated by cocultivation of blood leukocytes from chronically infected animals and cultures of the tumour cells with human embryo fibroblasts (HEF). HSV and Gross murine leukaemia virus antigens were detected in tumour cells by immunofluorescence and radioimmunoassay, respectively, and HSV antigen by immunofluorescence also in cultures of tumour cells and in cells of the brains, spinal cords, livers and spleens of the animals. HSV antibody was demonstrable in the blood serum from chronically infected tumour-bearing mice in a titre of 32.

Interaction of human lymphocytes and viruses in vitro.

Interaction between phytohaemagglutinin (PHA) stimulated human lymphocytes and two DNA viruses (adenovirus type 5 and herpes simplex virus type 1) considered to be closely connected with lymphoid tissues has been studied. The fate of the same viruses was investigated also in non-stimulated separated lymphocytes for comparative purposes. To elicit this interaction infectivity titrations, immunofluorescent technique and electron microscopy were used. The production of viral antigens was investigated by complement fixation. It has been shown that in PHA-stimulated lymphocytes from peripheral blood of healthy donors adenovirus type 5 is capable to replicate in its infectious form. Prolongation of the interval between stimulation and infection of cells significantly influenced the dynamics of replication. Non-stimulated lymphocytes produced antigens but no infectious particles.

Investigation of the nature of Australia antigen. I: The absence of biological activity of Australia antigen in human blood leukocyte culture.

Purified and concentrated preparations of Australia antigen had no stimulating effect on leukocytes of human subjects under study when tested either on DNA-polymerase activity, 3H-thymidine uptake or chromosomal alterations. Moreover, in patients with chronic hepatitis and cirrhosis of the liver no correlation between antigenemia and chromosome aberrations in blood leukocyte cultures could be detected. On the other hand, a serum obtained from a virus hepatitis patient with Australia antigen in the blood was found to stimulate leukocyte cultures from one patient with Down's syndrome and antigenemia, one mentally retarded patient and three normal donors. This stimulating agent is obviously not associated with Australia antigen.