[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.

[A summary of the data about antigenic and genetic diversity of rabies virus circulating in the terrestrial mammals in Russia].

The data about antigenic and molecular-genetic diversity of the rabies virus circulating in Russia are reviewed. Based on our studies and the literature data analysis circulation of two phylogenetic virus groups in Russia was revealed: Arctic and cosmopolitic. The Arctic group includes the subgroups of proper Arctic and Arctic-like viruses; the cosmopolitic--Central Russian, Northeastern European and steppe. It was found that the division into subgroups corresponded to the geographic distribution of rabies viruses.

[Production of the monoclonal antibodies to the rabies virus nucleoprotein].

Five hybridomas secreting monoclonal antibodies (MAbs) for the nucleocapsid protein of the rabies virus were obtained through the fusion of the SP2/0 murine myeloma cells with splenocytes of BALB/c mice immunized with fixed rabies virus (CVS strain). All hybridomas secret MAbs of the IgG class that display different specificity to the nucleocapsids of rabies and rabies-related viruses. MAbs 2ell showed the specificity for the prevalent in Russia rabies viruses that are similar to commercially available anti-rabies conjugate.

[A new principle in the selection of vaccinal rabies virus based on quantitation of the expression of G protein, a major immunogen of rabies virus].

The paper describes a new principle of the selection of the rabies virus vaccine strain ERA-CB20M based on quantitation of the expression of glycoprotein, a major immunogen of rabies virus. There is a correlation between the level of glycoprotein expression and that of vaccine virus attenuation. The application of the new principle for vaccine virus selection will permit a safer and more immunogenic rabies vaccine to be prepared.

[Protective activity of panavir in experimental rabies infection].

Significant and reliable activity of panavir was shown in experiments on mice intramuscularly infected with the rabies virus. The animal protection depended on the dose of panavir and the treatment scheme. The reliable protection (30-45% of the mice, p<0.05-0.01) was observed only after the panavir intramuscular injection to the site of the virus inoculation (the infection atrium), but not after the systemic (intraperitoneal) administration. High titers of the virus-neutralizing antibodies (7.92-10.45 IU/ml), comparable to the antibody titers in the combined suspensions of the brain tissues (6.01-10.45% IU/ml), were detected in the combined sera of the survived animals treated with panavir. For the first time a quantitative method for determination of the virus-neutralizing antibody levels in IU/ml was used (the FAVN test, the gold standard for comparative evaluation of the level of the vaccinal and infective antirabies immunity).

Detection of type 2 herpes simplex virus in cells of spermatogenic epithelium in infected testes of guinea pigs.

We developed a model of herpetic orchitis in guinea pigs. Intratesticular inoculation of type 2 herpes simplex virus suspension results in infection of the testicular spermatocytes and spermatides. The possibility of viral infection dissemination from infected into intact testis is proven.

[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.

[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.

[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.

[Effectiveness of the interferon inducers ridostin and camedon in prevention and treatment of experimental alpha- and flavivirus infections]. / Effektivnost' induktorov interferona ridostina i kamedona v profilaktike i lechenii éksperimental'nykh al'fa- i flavivirusnykh infektsii.

Antiviral activity of rybamidil (virasol) and interferon inducers ridostine and camedone developed in Russia and introduced in practical medicine has been studied in mice with experimentally induced arboviral infections, flavivirus (tick-borne encephalitis-TBE, and yellow fever-YF) and alphavirus ones (Western and Eastern equine encephalomyelitis-WEE and EEE). Rybamidil injected subcutaneously proved to be ineffective both for prevention and treatment of these infections. Early interferon inducers ridostine and camedone (larifan) with a peak of interferon production 4 hours after injection, when used in doses of 5 and 150 mg/ kg, respectively, had a clear-cut prophylactic antiviral effect if injected 24 or 4 hours before infection. Ridostine had a distinct antiviral effect, providing up to 40% protection from TBE, YF, and WEE.

[The efficacy of a cultured rabies vaccine studied in guinea pigs preliminarily infected with a fixed rabies virus]. / Izuchenie éffektivnosti kul'tural'noi antirabicheskoi vaktsiny na morskikh svinkakh v usloviiakh predvaritel'nogo zarazheniia ikh fiksirovannym virusom beshenstva.

In experiments of curative vaccination, carried out with the use of an experimental model similar to the current practice of treatment with antirabies preparations, the advantages of using tissue-culture rabies vaccine with immunogenic potency equal to 1.3 international units (I. U.) were shown. In these experiments the vaccine was introduced into guinea pigs infected with fixed rabies virus, the course of vaccination consisting of 14 daily injections. No correlation between the induction of virus-neutralizing antibodies and the immunogenic potency of tissue-culture rabies vaccine was established: the use of the vaccine with immunogenic potency equal to 0,3 and 1,3 I.U. had no essential influence on the level of antibody formation in the animals.

[The purification and concentration of the rabies virus by a diafiltration concentration method]. / Ochistka i kontsentrirovanie virusa beshenstva metodom diafil'tratsionnogo kontsentrirovaniia.

Purified concentrates of rabies virus were prepared by both microfiltration chromatography and diafiltration methods. The diafiltration concentrate had a high level of protective activity and was not inferior to the microfiltration chromatographic one. The infectious activity was better retained after diafiltration, the chromatographic purity of both preparations was similar. It is concluded that diafiltration presents a good alternative to the currently used methods for purification and concentration of viruses.

[The interaction of monoclonal antibodies to the structural proteins of the Vnukovo-32 vaccinal virus with other viruses of the rabies group]. / Vzaimodeistvie monoklonal'nykh antitel k strukturnym belkam vaktsinnogo virusa Vnukovo-32 s drugimi virusami gruppy beshenstva.

The interaction of 7 monoclonal antibodies (MCA) to the nucleocapsid complex and 3 MCA to protein G of the vaccine virus Vnukovo-32 with 33 and 27 members of the rabies virus group, respectively, was studied. The indirect immunofluorescence test showed 7 MCA to the nucleocapsid complex to recognize 4 antigenic determinants (AD). Two MCA recognized the AD common for all the viruses under study. Individual ecological variants may be detected using other MCA. Three MCA to protein G had marked virus-neutralizing activity. MCA 1C5 neutralized all the rabies viruses under study. The other two MCA differed in the number of viruses they could neutralize. The viruses under study were divided into 4 groups depending on their interactions with certain MCA in neutralization test.

[The isolation and characteristics of hybridomas producing monoclonal antibodies to the structural proteins of the Vnukovo-32 strain of the rabies virus]. / Poluchenie i kharakteristika gibridom, produtsiruiushchikh monoklonal'nye antitela k strukturnym belkam virusa beshenstva, shtamm Vnukovo-32.

A series of hybridomas secreting monoclonal antibodies (MCA) to glycoprotein and nucleocapsid proteins of rabies virus strain Vnukovo-32 was selected as a result of fusion of splenocytes from immune BALB/c mice with cells of myeloma line Sp2/OAq14, screening and cloning by limiting dilution methods in semi-liquid agar. Four hybridomas secreted MCA to glycoprotein in high titres (5.0 x 10(5)-2.2 x 10(6)) and had marked virus-neutralizing and therapeutic properties. Eight hybridomas produced MCA to the nucleocapsid complex: five hybridomas secreted MCA of the G class in high titres (2.4 x 10(5)-1.6 x 10(6)) and three hybridomas secreted MCA of the M class in low titres.

[The efficacy of a cultured rabies vaccine studied on guinea pigs previously infected with the rabies street virus]. / Izuchenie éffektivnosti kul'tural'noi antirabicheskoi vaktsiny na morskikh svinkakh, predvaritel'no zarazhennykh ulichnym virusom beshenstva.

In this work materials on the development of an experimental model for the study of rabies vaccines are presented. The comparative study of different immunization schedules for vaccines with different protective potency has been carried out. Guinea pigs infected with street rabies virus, strain k, were used as an experimental model. As shown in this investigation the optimum method of infecting the animals with strain k was intramuscular injection causing 50% mortality among the animals, the incubation period lasting 10-24 days. Only those tissue-culture rabies vaccines which had activity equal to 1.0-1.3 I. U. and, when injected into the animals, ensured survival rate ranging from 57% to 76%, depending on the immunization schedule, were shown to possess protective potency. It should be pointed out that survival rate among the animals receiving the preparation according to the reduced schedules recommended by WHO was higher than among those immunized daily for 14 days. In all groups immune response was observed. Still in the animals receiving the preparation according to the reduced schedules a higher level of virus-neutralizing antibodies was registered. Thus, an experimental model capable of being used for the evaluation of the quality of existing and newly developed antirabies preparations was obtained. Besides, we believe it to be expedient to carry out the field trial of rabies vaccines with activity equal to 1.0-1.3 I. U., using the reduced immunization schedules.

[Experimental study of the regularity of T-killer induction as a result of anti-rabies immunization]. / Izucehnie v éksperimente zakonomernostei induktsii T-killerov pri antirabicheskoi immunizatsii.

Experiments in CBA, mice immunized with inactivated unconcentrated cultural rabies vaccine (CRV) demonstrated that a clone of T-effectors of memory was formed within 30-37 days after primary immunization which reacted by the secondary type in response to inoculation of subthreshold doses of the rabies antigen. This clone of immunological memory seems to remain in the quiescent state for very long periods (the observation period 406 days). The immune splenocytes from the donors which had been given subthreshold doses of the antigen at intervals of 60, 103, 180 and 406 days, protected 2-4 times more animals than those after primary immunization with a threshold dose of the antigen. A continuous state of cell-mediated immunity--T-effectors processing the protective properties in vivo--may be maintained by repeated threshold doses of antigen given at 7-day intervals. The observed patterns of the induction of T-effectors possessing protective properties in vivo allowed some principles for construction of the optimal schedule of rabies vaccination to be formulated.

[Abortive and recurrent rabies in dogs intracerebrally infected with the rabies street virus]. / Abortivnoe i vozvratnoe beshenstvo u sobak, zarazhennykh v mozg ulichnym virusom beshenstva.

In the experiments on dogs inoculated intracerebrally with the "paralytic rabies" variant of the "Yak" strain of street rabies virus we observed spontaneous recovery of 1 out of 5 sick animals. Rabies in this dog was verified by three isolations of the virus from the saliva during the disease and by an increase of antibody titres in the time course of to 6776. In another experiment, among 7 dogs developing rabies after intracerebral inoculation with the "convulsive rabies" variant of the virus one animal showed recurrent form of rabies. The diagnosis of rabies was confirmed by isolation of the virus from the saliva in primary disease as well as by virus isolation from the salivary glands and the brain of the dog dying with recurrent rabies.

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 daily immunization and antirabies gamma-globulin on the development of immune T-killers in inbred mice infected with the street rabies virus]. / Vliianie ezhednevnoi immunizatsii i antirabicheskogo gamma-globulina na formirovanie immunnykh T-killerov u lineinykh myshei, zarazhennykh ulichnym virusom beshenstva.

Daily administration of vaccine as well as combined use of antirabies gamma-globulin and vaccine were shown not to induce the production of immune T-killers possessing protective properties in adoptive transfer in mice pre-infected with street rabies virus in contrast to a single administration of a shock dose of antigen. Immune splenocytes protected 31-43% of mice infected with street rabies virus in a dose of 2-4 im LD50 and did not protect the animals infected with a dose of 12 im-LD50. After combined administration of immune splenocytes and an interferon inducer (double-stranded RNA) the additive protective effect was observed.

[Immunogenic properties of the liposomal form of the rabies vaccine]. / Izuchenie immunogennykh svoistv liposomal'noi formy antirabicheskoi vaktsiny.

The liposomal form of inactivated cultural whole-virion rabies vaccine was shown to induce formation of more intensive immunity than the vaccine without liposomes. These data have been obtained in immunity challenge by intracerebral inoculation of fixed virus as well as intramuscular inoculation of street rabies virus.