RESUMO
The aim of the present study was to determine whether lipoarabinomannan (LAM), in combination with Freund’s incomplete adjuvant (FIA), was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA) in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-γ production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-γ production in response to ConA and OVA (P < 0.05). Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-γ production by at least 50 and 25 percent, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis.
Assuntos
Animais , Bovinos , Anticorpos Antibacterianos/imunologia , Doenças dos Bovinos/prevenção & controle , Adjuvante de Freund/imunologia , Lipídeos/imunologia , Lipopolissacarídeos/imunologia , Mycobacterium avium/imunologia , Ovalbumina/imunologia , Paratuberculose/prevenção & controle , Formação de Anticorpos/imunologia , Doenças dos Bovinos/imunologia , Ensaio de Imunoadsorção Enzimática , Adjuvante de Freund/administração & dosagem , Hipersensibilidade Tardia/imunologia , Hipersensibilidade Tardia/veterinária , Imunidade Celular , Lipídeos/administração & dosagem , Lipopolissacarídeos/administração & dosagem , Mycobacterium avium/química , Ovalbumina/administração & dosagem , Paratuberculose/imunologiaRESUMO
The aim of the present study was to determine whether lipoarabinomannan (LAM), in combination with Freund's incomplete adjuvant (FIA), was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA) in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-γ production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-γ production in response to ConA and OVA (P < 0.05). Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-γ production by at least 50 and 25%, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis.
Assuntos
Anticorpos Antibacterianos/imunologia , Doenças dos Bovinos/prevenção & controle , Adjuvante de Freund/imunologia , Lipídeos/imunologia , Lipopolissacarídeos/imunologia , Mycobacterium avium/imunologia , Ovalbumina/imunologia , Paratuberculose/prevenção & controle , Animais , Formação de Anticorpos/imunologia , Bovinos , Doenças dos Bovinos/imunologia , Ensaio de Imunoadsorção Enzimática , Adjuvante de Freund/administração & dosagem , Hipersensibilidade Tardia/imunologia , Hipersensibilidade Tardia/veterinária , Imunidade Celular , Lipídeos/administração & dosagem , Lipopolissacarídeos/administração & dosagem , Mycobacterium avium/química , Ovalbumina/administração & dosagem , Paratuberculose/imunologiaRESUMO
Distemper virus causes a disease affecting minks with respiratory, gastrointestinal, neurological and skin symptoms and showing high morbidity and mortality, mainly among puppies. It is controlled through immunization, using vaccines that are supplied for mink use. The aim of this work was to determine the seroneutralization titer against the distemper virus at a mink farm in Argentina. The antibody kinetics obtained after vaccination in 27 adult animals, as well as the duration of colostrum-transferred antibodies in 10 puppies were determined. All vaccinated adult minks showed protective titers up to at least 3 months after vaccination, and 37.5% significantly reduced their antibody levels, 12 months after vaccination. Only 20% of the puppies showed protective levels of colostrum-transferred antibodies at the age of 7 weeks, while non-detectable levels of antibodies were found when puppies reached 11 weeks old. Vaccination performed in these puppies at the age of 13 weeks, elicited protective seroneutralization titers. These results show that vaccination induces a satisfactory humoral immune response in our environment, and support the convenience of vaccinating dams annually before the beginning of the breeding season. The vaccination plan in puppies is also discussed.
Assuntos
Cinomose/prevenção & controle , Vison/imunologia , Morbillivirus/imunologia , Vacinas contra Parainfluenza/imunologia , Animais , ArgentinaRESUMO
Paratuberculosis or Johne's disease is a chronic infectious disorder caused by Mycobacterium avium subsp. paratuberculosis (Map). The disease produces diarrhea and weight loss in cattle and other animal species, and it is characterized by granulomatous enteritis and lymphadenitis. Histopathology and in situ techniques can be used as a diagnostic test, but the performance of these methods was not previously compared. The aim of this paper was to evaluate the ability of immunohistochemistry and in situ hybridization to detect Map in formalin-fixed tissue samples from infected cattle. Samples (ileum or ileocecal lymph node) from four animals that had positive Map culturing, lesions and detectable acid fast bacilli, as well as from two control animals, were tested by immunohistochemistry and in situ hybridization. Immunostaining and positive hybridization were observed in areas with lesions from infected animal samples, inside the cytoplasm of macrophages, epithelioid and giant cells. Immunostaining was intense in three samples and weak in one, while hybridization was weak in all cases. In situ hybridization was positive in negative areas of tissues analyzed by immunohistochemistry, which could be related to spheroplast detection as it was previously described for this method. Control samples resulted negative by these two methods. Both techniques were able to detect Map in formalin fixed and paraffin embedded tissues, however immunohistochemistry produced higher intensity staining and was easier to perform. Therefore, we believe that immunohistochemistry and in situ hybridization to be useful for the post-mortem diagnosis and research of Paratuberculosis.
Assuntos
Íleo/microbiologia , Imuno-Histoquímica/veterinária , Hibridização In Situ/veterinária , Linfonodos/microbiologia , Mycobacterium avium subsp. paratuberculosis/isolamento & purificação , Paratuberculose/diagnóstico , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/microbiologia , Paratuberculose/microbiologia , Álcool Feniletílico/análogos & derivados , Sensibilidade e EspecificidadeRESUMO
Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) causes Johne's disease, a chronic and fatal enteritis in ruminants. In the last stage of the disease, antibody titres rise and levels of interferon-gamma decrease, suggesting that the host-immune response is switching from a T helper 1 (Th1) to a Th2 profile. In infected cattle, the membrane protein p34 elicits the predominant humoral response against M. paratuberculosis. To map the B-cell epitopes of this antigen, affinity-purified bovine antibodies against the carboxy-terminal region of p34 were used to screen a 12-mer phage display library. Several phage clones carrying peptides resembling fragments of p34 were affinity selected. Based on the predicted amino acid sequence, peptides were chemically synthesized, which demonstrated reactivity with serum from naturally infected and p34-vaccinated cattle. Immunization of mice with these peptides elicited an anti-p34 antibody response. Two B-cell epitopes were identified and characterized. Based on the reactivity and the type of immune response elicited, epitope A was determined to be conformational, whereas epitope B was demonstrated to be sequential. Both epitopes were shown to be present in p34 proteins from M. avium ssp. avium or M. paratuberculosis but absent from M. intracellulare, the other member of the M. avium complex. Furthermore, both epitopes were mapped to regions of p34 that display high variability when compared to homologous proteins from other mycobacterial species of public and animal health importance. We hypothesize that these variable regions of p34 may play a role in the immunobiology of M. paratuberculosis infections.
Assuntos
Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Epitopos de Linfócito B , Mycobacterium avium subsp. paratuberculosis/imunologia , Sequência de Aminoácidos , Animais , Bovinos , Dados de Sequência Molecular , Paratuberculose/imunologiaRESUMO
Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.
Assuntos
Variação Antigênica , Antígenos de Bactérias/imunologia , Doenças dos Bovinos/microbiologia , Leptospira interrogans/imunologia , Doença de Weil/veterinária , Testes de Aglutinação , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Especificidade de Anticorpos , Antígenos de Bactérias/isolamento & purificação , Argentina , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/isolamento & purificação , Vacinas Bacterianas , Western Blotting , Bovinos , Doenças dos Bovinos/sangue , Eletroforese em Gel de Poliacrilamida , Leptospira interrogans/classificação , Leptospira interrogans/isolamento & purificação , Padrões de Referência , Coloração e Rotulagem , Doença de Weil/sangue , Doença de Weil/microbiologiaRESUMO
Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.(AU)
Assuntos
Animais , Bovinos , Estudo Comparativo , RESEARCH SUPPORT, NON-U.S. GOVT , Variação Antigênica , Antígenos de Bactérias/imunologia , Doenças dos Bovinos/microbiologia , Leptospira interrogans/imunologia , Doença de Weil/veterinária , Testes de Aglutinação , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Especificidade de Anticorpos , Antígenos de Bactérias/isolamento & purificação , Argentina , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/isolamento & purificação , Vacinas Bacterianas , Western Blotting , Doenças dos Bovinos/sangue , Eletroforese em Gel de Poliacrilamida , Leptospira interrogans/classificação , Leptospira interrogans/isolamento & purificação , Padrões de Referência , Coloração e Rotulagem , Doença de Weil/sangue , Doença de Weil/microbiologiaRESUMO
Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.
Assuntos
Animais , Bovinos , Variação Antigênica , Antígenos de Bactérias/imunologia , Doença de Weil/veterinária , Doenças dos Bovinos/microbiologia , Leptospira interrogans , Testes de Aglutinação , Especificidade de Anticorpos , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/isolamento & purificação , Argentina , Vacinas Bacterianas , Western Blotting , Doença de Weil/sangue , Doença de Weil/microbiologia , Doenças dos Bovinos/sangue , Eletroforese em Gel de Poliacrilamida , Leptospira interrogans , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/isolamento & purificação , Padrões de Referência , Coloração e RotulagemRESUMO
Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.
RESUMO
Foot and Mouth Disease Virus (FMDV) is one of the most feared animal virus and vaccination still has to be used in many countries. In previous reports, using a murine model, we studied the cellular basis of immune responses against FMDV and were able to show that they are atypical. In cattle, although complete protection may be attained after only one dose of killed virus vaccine, very little is known about protection against FMDV, except for antibody responses, but practically nothing concerning the cellular basis of their immune response. Moreover, since neutralizing titers do not always correlate with protection, the potency of vaccines in controlled by viral challenge. Our aim is to study cellular immune responses against FMDV, and to search for a correlate to protection. As a first step, 55 virgin cattle from a non endemic area (Patagonia) were divided into three groups: C: non immunized controls; HS: immunized with saponine containing vaccine; and EO: with oil emulsified vaccine. After vaccination, they were carried to an endemic area (Buenos Aires), where they were challenged with live FMDV. Animals were bled immediately before and 7 days after challenge, and their white blood cells and lymphocyte subpopulations were counted. All animals showed a marked neutropenia and eosinophilia, significantly higher in HS than in EO and C groups; both parameters were significantly better in the 2nd assay. Total lymphocyte counts were normal. Lymphocyte subpopulations were assessed by immunofluorescence using monoclonal antibodies: their proportions were normal and did not change during illness in group C. Several factors could have induced the observed eosinophilia and neutropenia: parasites, stress, saponine, others.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Aphthovirus/imunologia , Doenças dos Bovinos/prevenção & controle , Febre Aftosa/prevenção & controle , Vacinação/veterinária , Vacinas Virais/imunologia , Animais , Linfócitos B/química , Bovinos , Doenças dos Bovinos/sangue , Febre Aftosa/sangue , Imunidade Celular , Leucócitos/químicaRESUMO
Foot and Mouth Disease Virus (FMDV) is one of the most feared animal virus and vaccination still has to be used in many countries. In previous reports, using a murine model, we studied the cellular basis of immune responses against FMDV and were able to show that they are atypical. In cattle, although complete protection may be attained after only one dose of killed virus vaccine, very little is known about protection against FMDV, except for antibody responses, but practically nothing concerning the cellular basis of their immune response. Moreover, since neutralizing titers do not always correlate with protection, the potency of vaccines in controlled by viral challenge. Our aim is to study cellular immune responses against FMDV, and to search for a correlate to protection. As a first step, 55 virgin cattle from a non endemic area (Patagonia) were divided into three groups: C: non immunized controls; HS: immunized with saponine containing vaccine; and EO: with oil emulsified vaccine. After vaccination, they were carried to an endemic area (Buenos Aires), where they were challenged with live FMDV. Animals were bled immediately before and 7 days after challenge, and their white blood cells and lymphocyte subpopulations were counted. All animals showed a marked neutropenia and eosinophilia, significantly higher in HS than in EO and C groups; both parameters were significantly better in the 2nd assay. Total lymphocyte counts were normal. Lymphocyte subpopulations were assessed by immunofluorescence using monoclonal antibodies: their proportions were normal and did not change during illness in group C. Several factors could have induced the observed eosinophilia and neutropenia: parasites, stress, saponine, others.(ABSTRACT TRUNCATED AT 250 WORDS)