RESUMO
Immunization of BALB/c mice with Rickettsia prowazekii antigens, Bordetella pertussis toxin and Legionella pneumophila cytolysin induces the synthesis of IgM autoantibodies of different specificity. Among monoclonal antibodies, multispecific antibodies with a wide reactivity spectrum have been found to make up high percentage (30-80%). Monoclonal antibodies interact with different bacterial antigens and tissue substances. A hypothesis has been put forward that normally the injection of the antigen is followed by the appearance of antigen-nonspecific "immunological noise", including the synthesis of both tissue-specific and multispecific autoantibodies. Such antigen nonspecific "immunological noise" must have a certain threshold level which can be determined with the use of hybridoma techniques. This problem is particularly topical for bacterial antigens, as many of them are used in the development of vaccinal preparations, which may lead to an increase in the synthesis of autoantibodies and induce different autoimmune disturbances in the body.
Assuntos
Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos/imunologia , Animais , Anticorpos Monoclonais/isolamento & purificação , Reações Antígeno-Anticorpo/imunologia , Antígenos de Bactérias/imunologia , Autoanticorpos/biossíntese , Autoanticorpos/imunologia , Hibridomas/imunologia , Imunização/métodos , Imunoglobulina M/biossíntese , Imunoglobulina M/imunologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Studies on the typing of L. pneumophila strains of serogroup 1, isolated from patients and environmental objects, have been made with the use of monoclonal antibodies (McAb) to cytolysin. The results of the comparison of the specificity of our McAb with that of a commercial set McAb obtained from the USA make it possible to recommend preparations based on McAb to cytolysin for the detection of L. pneumophila pathogenic strains of serovar 1. The use of FITC- and peroxidase-labeled McAb to cytolysin permits the reduction of the time necessary for the diagnosis of Legionella infections and the detection of the antigen in a dose of 10 ng/ml.
Assuntos
Anticorpos Monoclonais , Citotoxinas/imunologia , Legionella/classificação , Especificidade de Anticorpos , Microbiologia Ambiental , Humanos , Legionella/isolamento & purificação , Legionella/patogenicidade , Sorotipagem/métodos , VirulênciaRESUMO
The study of the biological properties of monoclonal antibodies (McAb) to L. pneumophila cytolysin has been carried out. These McAb have been shown to possess no capacity for the in vitro neutralization of cytolysin and the protection of guinea pigs from aerosol infection with L. pneumophila. Still the protective effect of the McAb under study has been observed in experiments with the intraperitoneal infection of guinea pigs, which is indicative of the possibility, in principle, of involving humoral immunity into the protection of the body from Legionella infection.
Assuntos
Anticorpos Monoclonais/imunologia , Citotoxinas/imunologia , Legionella/imunologia , Administração Intranasal , Aerossóis , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/isolamento & purificação , Cobaias , Injeções Intraperitoneais , Doença dos Legionários/etiologia , Doença dos Legionários/prevenção & controle , Camundongos , Camundongos Endogâmicos BALB CRESUMO
R. prowazekii antigens have been tested with the use of monoclonal antibodies (McAb) to different epitopes of the microorganism. As revealed in these tests, McAb B4/4 and A-3/D, active against species-specific thermolabile antigen, interact with protein having a molecular weight of 90-120 KD. McAb C5/2, active against thermostable group antigen common with that of Rickettsia typhi, interact with LPS-like antigen having a molecular weight of 30 KD. Ultrastructural immunochemical studies have revealed that both R. prowazekii antigens are located on surface structures of rickettsiae, such as the microcapsule and cell wall.
Assuntos
Anticorpos Monoclonais , Antígenos de Bactérias/análise , Rickettsia prowazekii/imunologia , Antígenos de Superfície/análise , Coxiella/imunologia , Epitopos/imunologia , Técnicas Imunológicas , Microscopia Eletrônica , Peso Molecular , Rickettsia prowazekii/ultraestrutura , Rickettsia typhi/imunologia , Especificidade da EspécieRESUMO
A number of hybridomas to different R. prowazekii determinants were obtained by the hybridization of spleen cells of BALB/c mice immunized with R. prowazekii corpuscular and soluble antigens. Some of the monoclonal antibodies (McAb) reacted with R. prowazekii thermolabile species-specific protein and did not react with R. typhi antigens (McAb of batches B4/4 and A-D3). McAb C5/2 and A3/2 reacted with the group thermostable antigen, common for R. prowazekii and R. typhi. McAb to the species-specific thermolabile antigen belonged to IgG2a. The McAb thus obtained permit the identification of R. prowazekii and R. typhi and the solution of the problem of the intragroup differentiation of rickettsiae belonging to the typhus group.
Assuntos
Anticorpos Monoclonais/isolamento & purificação , Antígenos de Bactérias/imunologia , Epitopos/imunologia , Rickettsia prowazekii/imunologia , Animais , Anticorpos Monoclonais/análise , Antígenos de Bactérias/análise , Epitopos/análise , Hibridomas/imunologia , Técnicas Imunoenzimáticas , Camundongos , Camundongos Endogâmicos BALB C , Solubilidade , Especificidade da EspécieRESUMO
The fusion of spleen cells, taken from BALB/c mice immunized with the purified preparation of L. pneumophila cytolysin, with cells Sp2/0 and NP has been carried out. As a result, hybridoma cells producing IgG1, IgG3 and IgM antibodies to this protein have been obtained. All monoclonal antibodies (McAb) thus obtained react with L. pneumophila strain lysates in the precipitation test, while IgG3 and IgM antibodies react with erythrocyte diagnostic agents prepared from the lysate of L. pneumophila cells in the hemagglutination test. In the Western blot assay, McAb react with the 37 KD protein (cytolysin) and a number of other proteins from L. pneumophila cultures and L. pneumophila cell lysate, but do not react with the species-specific protein with a molecular weight of 29 KD, contained in the outer membrane of L. pneumophila, as well as with other species: L. bozemanii, L. dumoffii, L. longbeachae, L. micdadei. The possibility of using these McAb conjugated with FITC and peroxidase for the rapid diagnosis of Legionella infection is shown.
Assuntos
Anticorpos Monoclonais/biossíntese , Citotoxinas/imunologia , Legionella/imunologia , Animais , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/isolamento & purificação , Citotoxinas/isolamento & purificação , Eletroforese em Gel Bidimensional , Imunofluorescência , Testes de Hemaglutinação , Hibridomas/imunologia , Immunoblotting , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Especificidade da EspécieRESUMO
As the result of immunization of BALB/c mice with the commercial preparation of typhus vaccine and R. prowazekii corpuscular antigen, in 29.2% and 40.3% of cases (respectively) the appearance of hybridomas synthesizing monoclonal antibodies (McAb) to different autologous structures (skin and thymic epithelium, cell nuclei, conjunctive tissue structures and vascular endothelium) has been revealed. The McAb under test have proved to be IgM-autoantibodies. McAb M-6, active against the basal membrane of human skin and thymic epithelium, produce quite a definite picture of disturbances in the differentiation of epithelium and can be used for the diagnosis of dyskeratosis.
Assuntos
Anticorpos Monoclonais/isolamento & purificação , Antígenos de Bactérias/imunologia , Autoanticorpos/isolamento & purificação , Imunização/métodos , Rickettsia prowazekii/imunologia , Pele/imunologia , Timo/imunologia , Animais , Anticorpos Monoclonais/análise , Autoanticorpos/análise , Vacinas Bacterianas/imunologia , Membrana Basal/imunologia , Epitélio/imunologia , Humanos , Hibridomas/imunologia , Imunoglobulina M/análise , Imunoglobulina M/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB CRESUMO
By the indirect immunofluorescence method it was shown to which epithelial thymus structures monoclonal antibodies (mAT) reacting with the different epidermal structures are directed. These mAT related to the autoantibodies were obtained earlier, as a result of lymphoid cells polyclonal activation, by the immunization of BALB/c mice with streptococcal group A nonspecific protein antigens of the cell wall. It was shown that mAT A6/1, reacting with the basal layer of the skin epithelium are directed to the epithelium of the cortical and medullar thymus zones, which is regarded as the so called endocrinal epithelium. These mAT, by the study with immunoblotting method, react with the protein of mV SOkD, B5/1 mAT to the skin epithelium, on the thymus sections react with the single cells around the Hassel bodies.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Bactérias/imunologia , Autoanticorpos/imunologia , Streptococcus pyogenes/imunologia , Timo/imunologia , Animais , Epitélio/imunologia , Imunofluorescência , Humanos , Immunoblotting , Camundongos , Camundongos Endogâmicos BALB C , Pele/imunologiaRESUMO
The activity of B. pertussis toxin has been tested in the continuous culture of CHO (Chinese hamster ovary) cells. The in vitro method of testing B. pertussis toxin is rapid, highly sensitive and specific. The unit of activity of B. pertussis toxin is higher than in mouse tests by several orders. The specificity of the action of B. pertussis toxin on CHO cells has been confirmed by the test of the neutralization of the toxicity effect with antiserum.