[Monoclonal antibodies to rubella virus glycoprotein E1].

AIM: To obtain monoclonal antibodies (MCAs) to glycoprotein E1 of rubella virus, to assess their immunochemical characteristics and ability to use fluorescent MCA for rapid identification of rubella virus. MATERIALS AND METHODS: Rubella virus strain C-74 (Moscow), vaccine strains "Orlov" (Saint-Petersburg), Wistar RA 27/3 (USA) as well as strain Judith (Germany) were used. Viral antigens were obtained using diploid cells L-68 and cell lines VNK-21-F and Vero E6. MCAs were produced by conventional method and their isotype was determined: Immunoblotting, immunoenzume assay (IEA), hemagglutination inhibition assay (HIA) and immunofluorescence assay (IFA) were performed. RESULTS: Five monoclonal antibodies--Kh-252.1, Kh-347.2, Kh-183.3, Kh-214.4, Kh-187.5--to antigens of rubella virus strain C-74 were obtained. Isotypes of these antibodies were determined and their reactivity with native and denaturated antigens of other strains ("Orlov", Wistar RA 27/3, Judith) was characterized. IEA showed that all MCAs interacted with rubella virus glycoprotein E1 at high titers ranging from 1/1600 to 1/200,000. Immunoblotting demonstrated that 4 MCAs (Kh-252.1, Kh-347.2, Kh-183.3, Kh-214.4) had aforementioned feature. MCAs inhibited hemagglutinating activity of Judith strain in titer from 1/16 to 1/1024 in HIA. FITC conjugate of MCA Kh-347.2 (most sensitive variant) allowed to detect rubella virus in infected Vero E6 cells after 24 hours since infection, whereas FITC conjugates of 3 MCAs (Kh-183.3, Kh-214.4, Kh-187.5)--after 72 hours since infection. CONCLUSION: Use of FITC conjugates of MCAs is a perspective tool for identification of rubella virus glycoprotein E1 in infected cell cultures and nasopharyngeal swabs.

[Expression of receptors for human alpha- and gamma-interferons on the surface of peripheral blood mononuclear cells in viral infections]. / Ekspressiia retseptorov dlia chelovecheskikh alpha- i gamma-interferonov na poverkhnosti mononuklearnykh kletok perifericheskoi krovi pri virusnykh infektsiiakh.

Expression of receptors on peripheral blood mononuclears of donors and patients with viral infections during therapy was studied using FITC-labeled monoclonal antiidiotypical antibodies with the "internal image" of human alpha- and gamma-interferons and monoclonal antibodies to these interferons. Activation of the immune system caused by infection modifies the expression of interferon receptors and leads to appearance of membrane-bound interferons, mainly gamma-interferon.

[The protective effect of monoclonal anti-idiotypic antibodies in experimental herpes infection]. / Protektivnyi éffekt monoklonal'nykh antiidiotipicheskikh antitel pri éksperimental'noi gerpeticheskoi infektsii.

The protective effect of monoclonal anti-idiotypic antibodies m(anti-ID)Abs, simulating the biologic effects of human alpha interferons, were studied in male guinea pigs with genital herpes simplex. Therapeutic effect of m(anti-ID)Abs daily applied as liquid dosage form and as suppositories+ was assessed. Experiments demonstrated that m(anti-ID)Abs in liquid dosage form to a different measure decreased the severity of urogenital infection in guinea pigs and that m(anti-ID)Abs-25 completely prevented the development of grave infection.

[Monoclonal antibodies to Chlamydia psittaci: their isolation, characteristics and use]. / Monoklonal'nye antitela k Chlamydia psittaci: poluchenie, kharakteristika i primenenie.

A panel of 22 hybridomas producing monoclonal antibodies (McAb) to C. psittaci was obtained. 15 hybridomas produced IgG1 antibodies, 4 hybridomas produced IgM antibodies and 3 hybridomas produced IgG2b, IgG3 or IgA antibodies. IgG1 antibodies and 2 IgM antibodies did not bind complement in the complement fixation test. All McAb were reactive in the enzyme immunoassay and the indirect immunofluorescence test and did not precipitate specific antigens. Peroxidase conjugates on the basis of McAb effectively detected Chlamydia antigen, prepared from the crude suspension of chick embryo yolk sack infected with different strains of C. psittaci and C. trachomatis, in different modifications of EIA.

[Monoclonal anti-idiotypic antibodies simulating the biological effects of human alpha-interferons]. / Monoklonal'nye antiidiotipicheskie antitela, imitiruiushchie biologicheskie éffekty chelovecheskikh al'fa-interferonov.

The method of somatic hybridization was used to generate a panel of hybridomas producing monoclonal anti-idiotypic antibodies (mono-Ai-Ab) imitating biological effects of human alpha-interferons (hIF-alpha). Induction of syngeneic anti-idiotypic antibodies in BALB/c mice was achieved with monoclonal antibodies (MCA) IF-39 capable of neutralizing three kinds of hIF-alpha (lymphoblastoid, leukocyte, genetic-engineering). The screening of mono-Ai-Ab was done by determinations of antiviral activity (AV-activity) of supernatants from growing hybrid cell cultures caused by the cytopathic effect of 10-100 doses of mouse encephalomyocarditis virus (MEMC) by a micromethod in Vero cells grown in 96-well plates. Mono-Ai-Ab were found to neutralize MCA IF-39 and not to bind with immunosorbent of staphylococcal reagent containing protein A and BALB/c mouse immunoglobulins. It was shown that mono-Ai-Ab possessed AV-activity against MEMC and vesicular stomatitis viruses and were not inferior in this activity to commercial preparations of leukocyte IF-alpha. Mono-Ai-Ab had tissue species-specificity triggering the mechanism of AV-activity in human and simian cells as well as bovine kidney cells (MDVK line) imitating hIF-alpha in this effect.

[The protective effect of hemagglutinin-specific monoclonal antibodies in experimental influenza infection]. / Protektivnyi éffekt gemaggliutininspetsifichnykh monoklonal'nykh antitel pri éksperimental'noi grippoznoi infektsii.

Experiments in mice showed a high protective effect of monoclonal antibodies (MCA) to influenza A/Krasnodar/101/59 (H2N2) virus hemagglutinin, possessing neutralizing activity in ovo. A 100% protective effect was observed upon intranasal administration of MCA Kp/101-3 48 hours before infection, and 90% effect upon administration of MCA 96 hours before infection. A 100% therapeutic effect was observed upon intranasal administration of MCA Kp/101-3 less than 24 hours postinfection and 70% therapeutic effect was achieved by administration of these MCA 48-72 hours postinfection.

[2-site competitive immunoenzyme analysis based on monoclonal antibodies for the detection and titration of antihemagglutinating measles antibodies]. / Dvukhsaitovyi konkurentnyi immunofermentnyi analiz na osnove monoklonal'nykh antitel dlia vyiavleniia i titrovaniia antigemaggliutiniruiushchikh korevykh antitel.

Using monoclonal antibodies (MCA) to measles virus (strain Leningrad-16) hemagglutinin, a competitive enzyme immunoassay (cEIA) was developed for the detection and titration of antihemagglutinating antibodies in human sera. Serum samples from children, collected in measles foci, were tested in cEIA, SP-EIA, HI, and PHA tests. The results evidence a high correlation of antihemagglutinin titres in cEIA and HI tests, the correlation coefficient of cEIA and PHA being 97.7%. The cEIA based on MCA to measles virus hemagglutinin is highly specific for the detection and titration of measles antihemagglutinins, it does not involve the use of simian erythrocytes. A significant advantage of this test system consists in the possibility of using unpurified antigen prepared from the infected cell lysate.

[Preparation and properties of monoclonal antibodies to influenza virus A/Krasnodar/101/59 (H2N2)]. / Poluchenie i svoistva monoklonal'nykh antitel k virusu grippa A/krasnodar/101/59 (H2N2).

Somatic hybridization produced a set of 6 mouse hybridomas producing monoclonal antibodies of G isotype to influenza A/Krasnodar/101/59 (H2N2) virus. MCA were characterized by solid phase enzyme-immunoassay, hemagglutination-inhibition test, and indirect immunofluorescence technique. According to the results of radioimmunoprecipitation, all 6 hybridomas produced MCA to hemagglutinin of influenza A/Krasnodar/101/59 (H2N2) virus.

Preparation of Hybridomas producing monoclonal antibodies against human interferon.

To prepare hybridomas secreting monoclonal antibodies (MoAb) against human alpha-interferon (alpha-IFN), BALB/c mice were immunized with IFN produced in Namalwa cells. Native alpha-IFN, as well as partially purified or on cellulose adsorbed alpha-IFN preparations were used for immunization. Seven hybridomas continuously secreting IgG against human alpha-IFN were prepared by fusion of splenocytes from immunized donors with the mouse myeloma cells. MoAb reacted in ELISA as well as in neutralization test with human lymphoblastoid, leukocytic and recombinant alpha-IFN.

[Development and practical use of new experimental models of the various forms of herpetic infection]. / Razrabotka i prakticheskoe ispol'zovanie novykh éksperimental'nykh modelei raznykh form gerpeticheskoi infektsii.

New experimental models of neurological herpes in cotton rats and genital herpes in male guinea pigs have been developed which are more adequate to the corresponding human diseases, and models of ophthalmic herpes in rabbits and guinea pigs have been improved. These models may be used for screening and evaluation of the effectiveness of drugs for herpes. A high activity against herpes of bromovinyldeoxyuridine and acyclovir has been verified, a marked therapeutic effect of Soviet monophosphates ara-A, ara-C, and original silur preparation in some forms of herpes infection has been demonstrated.

[Acute meningoencephalitis in children caused by herpes simplex virus type I]. / Ostrye meningoéntsefality u detei, vyzvannye virusom prostogo gerpesa I tipa.

In 14 observations of acute meningoencephalitis in children mainly under 3 years of the age, data pointing to an etiological role of herpes simplex type I virus, were obtained on laboratory examination. Six children died, and in the rest 8 children who survived gross residua were observed. The morphological picture showed extensive colliquation necroses in the parietal, temporal, and less frequently, in occipital lobes and the Varolian pons. In 4 cases out of 6 intranuclear inclusions were detected. The incidence of herpetic encephalitis was 16%.

[Methods for the rapid diagnosis of herpes simplex using the indirect hemagglutination reaction]. / Metody bystroi diagnostiki gerpesa prostogo s pomoshch'iu reaktsii nepriamoi gemaggliutinatsii.

A method for preparation of an erythrocyte immunoglobulin herpes preparation for the indirect hemagglutination test (IHA) is described. A high specificity and sensitivity of this test and the preparation has been demonstrated. The results of the use of IHA with this preparation for the detection of herpes simplex virus in specimens from patients are presented.

[Production of an erythrocyte preparation for detection of herpes antibody in the hemagglutination reaction]. / Poluchenie éritrotsitarnogo preparata dlia vyiavleniia protivogerpeticheskikh antitel v reaktsii gemaggliutinatsii

A method for obtaining an erythrocyte diagnostic preparation for detection and titration of herpetic antibody in the passive hemagglutination test is reported. The use of this preparation for quantitative assay of herpetic antibody in sera from herpes simplex patients detected antibody in 100% of cases. Considerably less frequent findings of herpetic antibody were in human sera from diseases of other etiologies and in donor sera. Titers of antibody detected in the PHA test were 2--10 times higher than in the CFT. However, no differences between the PHA test and the CFT were found in per sent of seroconversions and in increase of antibody titers. The method is simple, sensitive and easy to perform in any clinical laboratory.