RESUMO
The outer membranes of Francisella tularensis were studied. The membranes were identified morphologically, immunologically and biochemically. They contained 12-20% of protein, 15-30% of carbohydrates, up to 40% of lipids. The main integral proteins of the outer membranes were the 47, 43, 17 and 12 kD proteins. The main protein 63 kD was not integral. The lipopolysaccharides isolated from the outer membranes and acetone-dried cells did not possess the protective properties in experimental tularemia. The preparations of outer membranes possessed the protective properties for mice infected with the virulent strain 503. Chitosan amplified the protective properties of outer membranes.
Assuntos
Vacinas Bacterianas , Quitina/análogos & derivados , Francisella tularensis/metabolismo , Antígenos de Bactérias/imunologia , Western Blotting , Membrana Celular/imunologia , Membrana Celular/metabolismo , Quitina/metabolismo , Quitosana , Eletroforese em Gel de Poliacrilamida , Francisella tularensis/imunologia , Soros Imunes , Imunização , Imunoeletroforese , Focalização Isoelétrica , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/metabolismoRESUMO
LPS-protein complex containing proteins of 15 kD, 17 kD and 19 kD was isolated from F. tularensis outer membrane by solving with sodium deoxycholate with the subsequent gel filtration on Sephacryl S-200. Protein of 17 kD constituted the main protein component of the complex. The LPS-protein ratio of this complex was 1:1. Proteins contained in LPS-protein complex have mainly the alpha-spiral structure. In the absence of detergent these proteins and LPS formed micelles with molecular weight exceeding 10(7) D. LPS-protein complex was shown to have a protective effect in mice infected with F. tularensis virulent strain 503.
Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Francisella tularensis/metabolismo , Lipopolissacarídeos/metabolismo , Animais , Cromatografia em Gel , Dicroísmo Circular , Eletroforese em Gel de Poliacrilamida , Camundongos , Micelas , Espectrofotometria InfravermelhoRESUMO
F. tularensis lipopolysaccharide (LPS) was studied with the use of monoclonal antibodies (McAb) having protective properties. The binding site of these McAb (IgG2a) is localized on the O-chain of LPS. In contrast to LPS isolated from vaccine strain 15, LPS isolated from F. tularensis cells in the R-form has no O-chains and does not interact with McAb.
Assuntos
Anticorpos Antibacterianos/análise , Anticorpos Monoclonais/análise , Especificidade de Anticorpos , Epitopos/análise , Francisella tularensis/imunologia , Lipopolissacarídeos/imunologia , Animais , Anticorpos Antibacterianos/isolamento & purificação , Anticorpos Monoclonais/isolamento & purificação , Sítios de Ligação de Anticorpos/imunologia , Epitopos/isolamento & purificação , Francisella tularensis/patogenicidade , Hibridomas/imunologia , Técnicas Imunológicas , Lipopolissacarídeos/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Inoculações Seriadas , Virulência/imunologiaRESUMO
The enzyme immunoassay and immunoblotting were used for the study of the serological activity of different mycobacterial antigens and the spectrum of antibodies to them in patients with different forms of tuberculosis and healthy persons. Antibodies in patients' sera were shown to bind antigens with different molecular weight. The level and spectrum of antibodies to purified protein fraction I made it possible to differentiate between patients with various forms of tuberculosis and healthy persons.
Assuntos
Antígenos de Bactérias/imunologia , Mycobacterium tuberculosis/imunologia , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/isolamento & purificação , Humanos , Immunoblotting/métodos , Técnicas Imunoenzimáticas , Peso Molecular , Tuberculoma/imunologia , Tuberculose Pulmonar/imunologiaRESUMO
The preventive activity of five monoclonal antibodies (McAb) in experimental tularemia was evaluated. McAb produced by hybridoma FB11-k (IgG2a), specific to F. tularensis lipopolysaccharide, prevented the death of mice and guinea pigs infected with F. tularensis virulent strain 503 of the holarctic subspecies.
Assuntos
Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos/imunologia , Francisella tularensis/imunologia , Lipopolissacarídeos/imunologia , Tularemia/prevenção & controle , Animais , Anticorpos Monoclonais/isolamento & purificação , Avaliação Pré-Clínica de Medicamentos , Francisella tularensis/patogenicidade , Cobaias , Hibridomas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Fatores de Tempo , Tularemia/imunologia , Virulência/imunologiaRESUMO
The protective properties of the preparation of F. tularensis outer membranes (OM), obtained from F. tularensis vaccine strain 15, were studied in experiments on hamadryas baboons challenged subcutaneously with F. tularensis virulent strain Schu (nonarctic subspecies). The subcutaneous immunization with the OM preparation prevented the development of clinically pronounced infection in more than 70% of the monkeys challenged with F. tularensis strain Schu in a dose of 787 live microbial cells 30 days after immunization. Antibody titers determined in the immunized monkeys with the use of the agglutination test (AT) and the passive hemagglutination test (PHAT) were usual in minimal diagnostic limits (1:80 for AT and 1:320 for PHAT) and did not significantly rise by day 20 after immunization. In all intact animals infected with F. tularensis strain Schu the development of the infectious process was registered, which was accompanied by a rise in temperature exceeding 39.5 degrees C and a rise in the titer of specific antibodies.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Francisella tularensis/imunologia , Papio/imunologia , Animais , Anticorpos Antibacterianos/sangue , Avaliação Pré-Clínica de Medicamentos , Feminino , Francisella tularensis/isolamento & purificação , Francisella tularensis/patogenicidade , Imunização , Masculino , Doenças dos Macacos/imunologia , Doenças dos Macacos/microbiologia , Doenças dos Macacos/patologia , Doenças dos Macacos/prevenção & controle , Fatores de Tempo , Tularemia/imunologia , Tularemia/microbiologia , Tularemia/patologia , Tularemia/prevenção & controle , VirulênciaRESUMO
Subcutaneous immunization, made in a single injection, with outer membrane preparations obtained from F.tularensis vaccine strain 15 and virulent strain A'Cole results in intensive immunity to tularemia in guinea pigs, ensuring the protection of 60-100% of the animals within a month after challenge with F.tularensis virulent strain 503 in a dose of 1,000 DCL. The development of protective effect induced by F.tularensis outer membranes can be observed during the first 24 hours and reaches its maximum by days 15-21 after immunization.