Local and Systemic Functional Responses of Mouse Macrophages to Intravaginal Infection with Type 2 Herpes Simplex Virus and Vaccination.

Activity of cathepsin D and phagocytosis of macrophages from vaginal lavage fluid, peritoneal exudation, and spleen were studied in mice of sensitive (DBA/2) and resistant (BALB/c) lines after intravaginal infection with type 2 herpes simplex virus and vaccination. Activity of cathepsin D and intensity of phagocytosis (irrespective of the macrophage source) and their ratio in BALB/c mice in early terms after infection were close to the control levels taken as a unit. In DBA/2 mice, these parameters and their balance were shifted and changes in cathepsin D activity depended on the time after challenge. Activities of cellular and extracellular cathepsin D increased sharply on day 1 postinfection under conditions of local virus interaction with the vaginal mucosa and activation of the pathological process. Later, after generalization of the infection, activity of cathepsin D decreased, while phagocytosis increased in all the studied macrophage populations. Vaccination corrected the cathepsin D/phagocytosis imbalance and created conditions for rapid elimination of the virus.

[Vaccines as an approach to the immunocorrection in herpetic infections].

Development of vaccines for immunologic correction in herpetic infections is an important problem that raises a growing concern worldwide. The data on the experimental studies of the efficacy of an inactivated whole-virion vaccine against herpes simplex viruses types 1 and 2 (HSV-1 and -2) using an animal model are discussed. The results of the multiyear application of the vaccine to ophthalmology and dermatology practice are also presented. The results unambiguously show a high efficacy of the vaccine in the prevention of recurrences of the infections based on activation of specific T-cell response. A live vaccine against the varicella zoster virus (VZV) was developed for control of the infection in children. For the cytomegalovirus (CMV) infection in adults, inactivated whole-virion vaccines are at the stage of development. An important part of the study addresses a combined application of the inactivated vaccines with immunomodulators.

[Immunomodulators and specific inactivated vaccines in urgent prophylaxis under experimental arboviral infection].

A reliable protective activity of the home-manufactured immunomodulators (ridostin, polyribonate glucosemuramyl-dipeptide, Mylife, and peptidoglycane-160) was detected in mice. The mice were infected with the equine eastern encephalomyelitis virus (EEEV, an alphavirus), or with the tick-borne encephalitis virus (TBEV), or the yellow fever (YF) virus (both flaviviruses). The effect of the urgent vaccination reliably increases when the vaccination is combined with the immunomodulators listed above. Under the alphavirus infection, the combined effects of the vaccine and ridostin were accompanied with increased specific humoral and cellular immune response (virus-specific antibodies and adoptive transfer of immune lymphocytes). The combined application of the specific vaccine and ridostin can be recommended for clinical trials of TBE in the foci of Infection.

[Efficiency of coadministration of immunomodulators and vaccine in an experiment on tick-borne encephalitis].

Experiments on a tick-borne encephalitis (TBE) model in CBA and BALB/c mice demonstrated that immunomodulators (ridostin, polyribonate, and peptidoglycan-160) and a specific vaccine against TBE were significantly effective in increasing the level of a protective effect and life expectancy in the experimental group as compared to the control group. The findings allow one to recommend the immunomodulator ridostin in combination with the inactivated vaccine for the emergency prophylaxis of TBE in its virus-infected subjects in the foci of infection.

[Combined effects of immunomodulators and specific inactivated vaccines against alpha- and flavivirus infections].

The combined effects of immunomodulators, such as ridostin, polyribonate, glucose muramyl dipeptide, and peptidoglycan-160, and specific vaccines on survival of mice with alpha- (eastern equine encephalitis) and flavivirus (tick-borne encephalitis (TBE) infections were found to be significant. In alphavirus infection, the combined effects of the specific vaccine and ridostin were accompanied by increases in specific humoral immunity (specific antibodies) and cellular immunity (adoptive transfer of immune lymphocytes). The concurrent use of the specific vaccine and the immunomodulator ridostin is recommended in clinical trials of TBE in the foci of infection.

[Inactivated herpes simplex virus types 1 and 2 divaccine as an agent for effective immunoprophylaxis of recurrent genital herpes].

Prevention of recurrent genital herpes with the inactivated herpetic divaccine Vitaherpavac against herpes simplex virus types 1 and 2 has a number of advantages over the most commonly used symptomatic therapy: it ceases or significantly reduces the number of recurrences and accordingly prolongs a relapse-free interval, abolishes viremia and the manifestations of clinical symptoms of recurrences, induces no dependence to the vaccine. Coadministration of the Vitaherpavac vaccine and the immunomodulator Giaferon has been shown to have some advantage over vaccination only. The new formulation of the agent as suppositories (per rectum) not only enhances the immunogenicity and protective properties of the vaccine, but also reduces the frequency of its application and makes more convenient for patients to use.

[Vitaherpavac is the first Russian herpes simplex virus vaccine obtained on the Vero B continuous cell line].

Vitaherpavac, a dry inactivated herpes simplex virus (HSV) culture vaccine, has been obtained, by using the Vero B continuous cell line as a substrate for accumulation of herpes simplex virus types 1 (US strain) and 2 (VN strain). Vitaherpavac and the similar vaccine Herpovax made by the Research Institute of Vaccines and Sera, Saint Petersburg (for which preparation a primary trypsinized chick embryo cell culture used as a substrate for accumulation of HSV types 1 and 2), underwent comparative clinical trials. The tolerability and therapeutic effectiveness of the vaccine were tested in patients diagnosed as having chronic frequently recurring herpes. The trials have yielded positive results that suggest that it is expedient to introduce of the new vaccine Vitaherpavac into practice to treat chronic recurrent herpetic infection of various localizations. Vitaherpavac has been registered in the Russian Federation and permitted for medical application.

Functional activity of peritoneal macrophages from sensitive and resistant mouse strains during intravaginal infection with herpes simplex virus type 2 and mucosal vaccination.

In the early period after intravaginal infection with herpes simplex virus type 2 (2 h), macrophages from sensitive DBA/2 mice were characterized by higher capacity to engulf the antigen, decreased function of the lysosomal apparatus, lower activity of cathepsin D, and reduced oxygen metabolism compared to cells from resistant BALB/c mice. Mucosal vaccination with herpes vaccine and hyaluronic acid promoted the increase in functional activity of macrophages and improved survival of sensitive mice (by 60%).

[Antiviral activity of Bacillus intermedius RNAase in guinea-pigs and rabbits infected with outdoor rabies virus].

Single injection of Bacillus intermedius RNAase in a dose of 5 mg/kg could protect 40 and 50-70% of the outdoor rabies virus-preinfected guinea-pigs and rabbits, respectively. In the control group there were 100 and 75-100% deaths of the RNAase-untreated guinea-pigs and rabbits, respectively. Animal protection was observed only when RNAase was injected into the site of viral administration. The intramuscular injection of RNAase, other than the site of viral administration failed to protect the infected animals. The efficacy (75%) of RNAase injected into the rabbits was similar 1 and 24 hours after animal infection.

[Herpesvirus infection in patients with chronic glomerulonephritis].

Virological examinations of blood, urine and saliva in 75 patients with chronic glomerulonephritis (CG) revealed, in 95% of them, herpes-virus infections caused by herpes simplex virus, type 1 (34.4%), herpes simplex virus, type 2 (2.6%) and cytomegalovirus (12%) or mixed infections (46%). The infection rate in the control group of children without renal pathology was reliably lower. A majority of CG patients (94%) had a diagnostically significant level of antiherpetic antibodies, class IgG, which also evidence to chronic herpes-virus infection.

[The antiviral activity of RNAse Bacillus intermedius in experiments with mice preinfected with street rabies virus]. / Protivovirusnaia aktivnost' RNKazy Bacillus intermedius v éksperimentakh na myshakh linii CBA, predvaritel'no zarazhennykh ulichnym virusom beshenstva.

RNAse Bacillus intermedius, when administered once and according to 11 repeated experiments, protected the preliminarily infected CBA mice with street rabies virus (protection of 40-67%; p < 0.01-0.001). A reliable protection of Animals was registered only when RNAse was administered intramuscularly at the virus introduction spot; it was not effective, when the bacterial RNAse was injected in the brain, vein, under the skin or in muscles of a non-infected extremity. Neither did it produce any suppressive effect on the vaccinal antirabic immunity.

[Antiviral activity of human recombinant gamma-interferon and recombinant hybrid protein of tumor necrosis factor-alpha-thymosin-alpha1 on models of herpes-virus and cytomegalovirus infection in vitro]. / Protivovirusnaia aktivnost' rekombinantnogo interferona-gamma cheloveka i rekombinantnogo gibridnogo belka alpha-faktor nekroza opukholei-timozin-alpha1 na modeliakh gerpes-virusnoi i tsitomegalovirusnoi infektsii in vitro.

The conducted studies showed a certain efficiency of gene-engineering preparation of IFN-gamma and TNF-T in virus infections caused by herpes simplex virus 2 (HSV-2) and cytomegalovirus (CMV) in cell cultures of human embryo fibroblast (HEF). The drugs have no viricidal action. IFN-gamma, when used according to the treatment scheme in vitro, proved to be more effective versus TMF-T both in HSV-2 and in CMV. It inhibited significantly the HSV-2 reproduction within the dilution range of 1:50 to 1:500. It was also effective, when used in CMV, within the dilution range of 1:5000 and lower, whereas TNF-T was effective within the range of 1:500 and lower as well as in 0.1 multiple infection. A significantly higher effect was ensured when the drugs were used for prevention. In HSV-2, IFN-gamma inhibited the virus reproduction, like in the treatment scheme, within the dilution range of 1:50 to 1:500, whereas TNF-T was effective in the range of 1:50. In CMV, the drugs' effect, when used for prevention, was similar to that observed in the treatment scheme. The highest inhibition values were registered for HSV-2, when it was used 24 hours before infection (IFN-gamma--2.25 Ig, dilution range of 1:50; TNF-2--1.0 Ig, dilution range of 1:50). IFN-gamma and TNF-2 exert a synergic action on different stages of virus reproduction. A reliable additive effect was ensured in prevention made 4 hour before infection by IFN-gamma and TNF-T only in experimental CMV infection.

[Polymorphism of herpes simplex virus type 1 and 2 DNA from laboratory strains and clinical material from patient with genital herpes]. / Polimorfizm DNK virusa prostogo gerpesa tipov 1 i 2 iz laboratornykh shtammov i klinicheskogo materiala ot patsientov s genital'nym gerpesom.

Polymerase chain reaction (PCR), that can amplify a fragment of the DNA-polymerase gene of 4 herpes viruses, i.e. herpes simplex viruses, type 1 (HSV-1), herpes simplex viruses, type 2 (HSV-2), Epstein-Barr virus and cytomegalovirus, was made use of to study the genetic polymorphism of HSV-1 and HSV-2 strains. The obtained amplicons were analyzed by the method of restriction-size fragments' polymorphism (RSFP) with restrictases Rsal, Taql and Hinfl. Four HSV-1 strains had an identical restriction profile. Strain G (HSV-2) also displayed the expected restriction profiles, however, contradictory results were obtained for strain BH (HSV-2): the restriction profiles with restrictases Hinfl and Rsal corresponded to HSV-2, and the restriction profile with Taql corresponded to HSV-1. The sequencing of appropriate fragments of strains G and BH revealed a dot-type mutation localized in Taql restriction site. The thus worked out PCR was used jointly with RSFP in the genotyping of 75 urogenital samples obtained from women with genital herpes who were treated at Moscow patient-care facilities. HSV-1 and HSV-2 were detected in 18 (24%) and 57 (76%) of samples, respectively. No changes were registered in the restriction profile for HSV-2 among the investigated samples and all of them had the restriction profile similar to that of strain G. The conclusion is that genital herpes associated with HSV-2 is genetically stable within its Moscow population.

[The antiviral activity of peptide immunomodulator "Gepon" in experimental infections caused by herpes simplex viruses types 1 and 2]. / Protivovirusnaia aktivnost' peptidnogo immunomoduliatora "Gepon" pri éksperimental'nykh infektsiiakh, vyzvannykh virusami prostogo gerpesa tipov 1 i 2.

Experimental data are reported on the antiviral activity of peptide immune-modulator "Gepon" in infections caused by herpes simplex virus (HSV), types 1 and 2, in vitro and in vivo. The drug proved to be non-toxic and did not possess any viricidal action. Its antiviral effect was registered in experiments with a multiple-infection cells' culture. The maximal effect was a 100-fold reduction of the viral titer when it was used in a concentration of 6.25 mcg/I as a preventive measure 24 hours before triggering the infection. The mentioned drug's effect was reliably higher than its use within a treatment scheme (1 hour after the infection onset). "Gepon" possessed the reliable protecting qualities (36% of protection with a mean increase of the infected mouse life by 1.9 days) in experiments with intraperitoneally infected mice (10 LD50/mouse of HSV, type 2), when the drug was administered in doses of 0.1 and 1 mcg/mouse.

[The antiviral activity of the peptide immunomodulator "Gepon" in experimental models of street rabies virus]. / Protivovirusnaia aktivnost' peptidnogo immunomodulatora "Gepon" v éksperimentakh na modeli ulichnogo beshenstva.

Intracerebral injections of the Gepon drug were shown to be non-toxic when administered to outbred white mice, 0.01-0.1 micrograms. The drug protects 30-40% of mice (p < 95-99%) in contamination by street rables virus. The animals' protection depends on a dose and drug application scheme. The most pronounced protection was observed in its intramuscular administration, 0.1 microgram in the virus locus (infection opening). It should be necessarily pointed out that such pronounced protection (40%; p < 0.01) was noted within the therapeutic scheme of drug administration after virus infected the animals. A new method is described, which is related with evaluating the antiviral drugs' quality, i.e. with registering the dynamics of morbidity and death of infected animals.

[Virus isolated in the human blood leukocyte culture and its interaction with hepatitis C and G viruses]. / Virus, vydelennyi na kul'ture leikotsitov krovi cheloveka, i ego vzaimootnoshenie s virusami gepatitov C i G.

Electrone-microscopic investigations are indicative of that the cultures of healthy donors, stimulated by phytohemagglutinin (PHA), can be successfully used to study the etiology of parenterally transmitted hepatitis. An electronic-microscopic study of a virus, isolated from the blood serum of a patient with hepatitis on the basis of the PHA-stimulated human leukocyte cultures and named a hepatitis leukocytic virus (HLV), enabled, by using the negative contrasting method, to detect viral particles of the hexagonal shape, sized 50-65 nm, with a coating divided by a 4-5 nm light space. Therefore, the HLV was described as belonging to the Flaviviridae family. RNA of the C hepatitis virus was detected in the K HLV strain stored, for 24 years, at the Museum of the Viruses Research Institute, Russian Academy of Medical Sciences, in a lyophilizated bed at -5 degrees C, however, an attempt to genotype the RNA failed. No RNA donor leukocytes were found in the materials of further passing of HLV by using the PHA-stimulated cultures, which can be explained by an inactivation of HLV at storage. No RNA of the C hepatitis virus was found in the above materials either, however, in 1999, DNA of the TT virus was detected at passing the strain, which indicates that the virus is widely spread in the population of healthy donors, whose lymphocytes are used preparing the blood leukocyte cultures.

[Therapeutic effect of gefin, a new antiviral drug, in experimental genital herpes ]. / Lechebnoe deistvie novogo protivovirusnogo preparata gefina pri éksperimental'nom genital'nom gerpese.

Antiviral activity of Gefin was studied in guinea pigs infected with herpes simplex virus (HSV-2). Up to 54.9% animals challenged with HSV survived after 7-day treatment with Gefin (3 local applications a day). The prospects of further trials of the drug in HSV genital herpes are discussed.

[Combined use of killed vaccines and immunomodulator ridostin for urgent prevention of epidemic stomatitis, Aujeszky disease and carnivore plague in experiment]. / Sochetannoe ispol'zovanie ubitykh vaktsin i immunomoduliatora ridostina dlia ékstrennoi profilaktiki iashchura, bolezni Aueski i chumy plotoiadnykh v éksperimente.

Results of experimental studies of mice and pigs infected with foot-and-mouth disease virus, minks infected with Aujeszky's disease virus, and dogs infected with canine distemper virus are described. In animals with foot-and-mouth disease and Aujeszky's disease, combined treatment with killed vaccine and immunomodulator Ridostin by the scheme of urgent prophylaxis (3 days before infection) caused 75% (foot-and-mouth disease) and 100% (Aujeszky's disease) prevention of animal death and development of generalized infection. The use of Ridostin by the scheme of urgent prophylaxis in a canine distemper infection focus arrested clinical symptoms of the disease in 50% of animals received immunomodulator. Clinical symptoms of canine distemper in the other dogs treated with immunomodulator were manifested in a mild form, and their appearance was delayed to 23-25 days after contact with infected animal.

[Mechanisms for the therapeutic effect of herpes polyvaccines in chronic ophthalmic herpes and genital herpes]. / Mekhanizm lechebnogo éffekta gerpeticheskoi polivaktsiny pri khronicheskom oftal'mogerpese i gerpes genitalii.

Commercial inactivated culture polyvaccine against herpes simplex viruses (types 1 and 2) developed at D. I. Ivanovsky Institute of Virology promoted cessation of viremia. During the first vaccination viremia coincided with appearance of a focal allergic test on the retina, which is proposed for the diagnosis of herpetic involvement of the posterior compartment of the eye. T-cellular immunity normalized after a course of vaccination. Experimental immunization of rats and vaccination of patients with chronic ophthalmic and genital herpes demonstrated the therapeutic activity of inactivated herpetic polyvaccine in suppositoria.

[Development of a suppository form of the interferon inducer poludan and potentiation of its action by hyaluronic acid]. / Razrobotka svechevoi formy induktora interferona poludan i potentsirovanie ego deistviia gialuronovoi kislotoi.

A promising dosage form of interferon inductor poludan has been developed for the treatment of patients with genital herpes and ophthalmic herpes: suppositoria. High interferon-producing and antiviral activity of poludan in suppositoria is potentiated by hyaluronic acid and antioxidants.