Assuntos
Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose/diagnóstico , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Mycobacterium tuberculosis/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Tuberculose/sangue , Tuberculose/imunologiaRESUMO
A representative collection was obtained containing 68 monoclonal antibodies (MAB) to Toxoplasma gondii antigens, which was characterized by the binding with the below fractions of tochizoites in the immune-enzyme assay (IEA) and immunoblotting (IB): membrane (MEM), somatic (water-soluble, SOM) and excretory-secretory (ES). Most of MABs were produced to MEM antigens (43), 6 MABs reacted with the somatic fraction, and 3 MABs reacted with both fractions. Two MABs to ES antigen were detected in the latter group. An analysis of MABs in concurrent IEA and IB revealed the immune-dominant proteins of the MEM and SOM fractions of antibodies to T. gondii tochizoites (p30 and p27, respectively). The presence of 2 non-overlapping antigenic determinants was shown for p30. Further research would detect MABs that could be used in the diagnosis of toxoplasmosis.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Protozoários/imunologia , Toxoplasma/imunologia , Animais , Membrana Celular/imunologia , Epitopos , Immunoblotting/métodos , Técnicas Imunoenzimáticas , Camundongos , Camundongos Endogâmicos BALB CRESUMO
A recombinant strain producing TBI protein (artificial protein containing HIV-1 B and T cell epitopes) was constructed on the base of attenuated Salmonella typhimurium SL7207 strain and BALB/c mice were immunized with this strain in a dose of 10(9) live cells orally or rectally. A single immunization with the recombinant salmonella strain induced humoral and cellular immune response to HIV-1; hence, this strain is a promising candidate for vaccine against HIV.
Assuntos
Vacinas contra a AIDS/imunologia , Genes Sintéticos , HIV-1/imunologia , Proteínas/imunologia , Salmonella typhimurium/imunologia , Vacinas contra a AIDS/administração & dosagem , Vacinas contra a AIDS/biossíntese , Administração Oral , Administração Retal , Animais , Anticorpos Antivirais/análise , Formação de Anticorpos , Infecções por HIV/imunologia , Infecções por HIV/prevenção & controle , Imunidade Celular , Técnicas Imunoenzimáticas , Camundongos , Camundongos Endogâmicos BALB C , Biossíntese de Proteínas , Proteínas/genética , Proteínas Recombinantes/biossíntese , Recombinação Genética , Salmonella typhimurium/genética , Baço/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/biossíntese , Vacinas Sintéticas/imunologiaRESUMO
A method for designing molecular constructions of combined artificial immunogens mimicking viral particles is proposed. Using this method, it is possible to expose antigenic determinants of any viral protein on the surface of such particles and to deliver plasmids containing genes encoding the synthesis of protein antigens to target cells. This approach was used to create the constructions of combined artificial immunogens inducing the production of specific antibodies to HIV-1 and to evaluate the extent and duration of B-cellular immune response depending on the way of antigen exposure to immunocompetent cells.
Assuntos
Vacinas Combinadas/genética , Vacinas Sintéticas/genética , Vacinas contra a AIDS/genética , Animais , Epitopos/genética , Produtos do Gene env/genética , Produtos do Gene gag/genética , HIV-1/genética , HIV-1/imunologia , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Plasmídeos/genética , Vacinas Combinadas/imunologia , Vacinas Sintéticas/imunologiaRESUMO
A gene encoding the dimeric form of single-chain antibody fragments (scFv) was designed on the basis of the artificial gene coding monomeric scFv to tick-borne encephalitis glycoprotein E. The sequence encoding histidine oligomer was added to 3'-ends of the genes encoding the monomeric and dimeric forms of scFv antibodies. Escherichia coli strains were constructed for the production of monomeric and dimeric antibodies. These antibodies were purified using Ni-NTA resin. The specificity of the purified monomeric and dimeric antibodies in the binding reaction with tick-borne encephalitis virus was shown by ELISA and Western-blot analysis. The identity of glycoprotein E epitope bound by monomeric and dimeric scFv and parental monoclonal antibodies E6B was confirmed by competitive immunoassay.
Assuntos
Anticorpos Antivirais/genética , Vírus da Encefalite Transmitidos por Carrapatos/imunologia , Anticorpos Antivirais/imunologia , Sequência de Bases , Western Blotting , Linhagem Celular , Primers do DNA , Ensaio de Imunoadsorção Enzimática , Humanos , Técnicas Imunoenzimáticas , Fragmentos de Imunoglobulinas/genética , Fragmentos de Imunoglobulinas/imunologia , Plasmídeos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologiaRESUMO
In this work the results of obtaining HBcAg-producing attenuated Salmonella strains, serovars S. enteritidis and S. typhimurium, and their comparative study is presented. As revealed in this study, attenuated S. enteritidis strain E-23 and S. typhimurium strain T-10, producing HBcAg, induce cell-mediated and humoral immune response to HBcAg after injected into anovals. After injection S. typhimurium strain T-10 induces a much higher titer of specific antibodies than S. enteritidis strain E-23. The level of specific antibodies induced by recombinant HBcAg seems to correlate with the capacity of salmonellae for survival inside macroorganisms.
Assuntos
Apresentação de Antígeno , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Salmonella enteritidis/imunologia , Salmonella typhimurium/imunologia , Animais , Vetores Genéticos/imunologia , Hepatite B/imunologia , Hepatite B/prevenção & controle , Antígenos do Núcleo do Vírus da Hepatite B/genética , Vacinas contra Hepatite B/genética , Vacinas contra Hepatite B/imunologia , Imunidade nas Mucosas , Camundongos , Camundongos Endogâmicos BALB C , Plasmídeos/genética , Recombinação Genética , Salmonella enteritidis/genética , Salmonella typhimurium/genética , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologiaRESUMO
Recombinant strains producing hepatitis B virus (HBcAg) core protein and chimeric core protein exposing on its surface the major immunogenic epitope of HBsAg (HBcAg-HBs) were constructed on the base of attenuated S. typhimurium SL 7202 strain. The resultant Salmonella strains produced proteins which were capable of self-assembly into virus-like particles and showed antigenic properties of both core and surface hepatitis B proteins. A single rectal immunization with recombinant S. typhimurium induced humoral and cellular immune response to HBcAg and HBsAg. Specific anti-HBcAg were detected in animal sera and intestinal tissues, which indicated the formation of specific mucosal immunity.
Assuntos
Antígenos do Núcleo do Vírus da Hepatite B/genética , Antígenos de Superfície da Hepatite B/genética , Salmonella typhimurium/genética , Animais , Epitopos/imunologia , Antígenos do Núcleo do Vírus da Hepatite B/biossíntese , Antígenos de Superfície da Hepatite B/biossíntese , Imunidade Celular , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica , Recombinação Genética , Salmonella typhimurium/imunologiaRESUMO
The nonstructural 36K protein of vaccinia virus (VV) mapped in HindIII-P and HindIII-J fragments of VV strain L-IVP has been expressed in E. coli as a fusion protein. The products of 36K gene preserved the antigen homology with the native protein 36K and the capacity to bind specific immunoglobulins of rabbit antiVV serum. The protective properties of 36K gene products and their joint effect with the immunodominant protein 35K were investigated. The non-structural 36K gene products showed no protective activity, but increased the production of specific antibodies in mice immunized with a mixture of both protein preparations, this increase being compatible with that observed after immunization with the inactivated virus preparations.
Assuntos
Escherichia coli/genética , Vaccinia virus/genética , Proteínas Virais/genética , Animais , Clonagem Molecular , Camundongos , Camundongos Endogâmicos BALB C , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Virais/genética , Vacinas Virais/imunologiaRESUMO
Three monospecific antisera to the major 35 kD (p35) surface protein of vaccinia and ectromelia viruses have been obtained. Two of them are obtained to p35 protein isolated by electrophoresis in the presence of sodium dodecylsulfate from the protein fractions of vaccinia virus, soluble in NP40 and NP40 with dithiothreitol (NP40 and DTT-fractions). The third serum is obtained to NP40-fraction of ectromelia virus, containing practically only p35 protein. The obtained antisera were compared in the reactions with the different fractions of viral proteins in two versions of solid phase radioimmunoassay. The effect of such reagents as sodium dodecylsulfate, NP40, 2-mercaptoethanol, ethanol on the antigenic properties of p35 protein from vaccinia virus is discussed.
