RESUMEN
Brucellosis, a zoonosis caused by Brucella, is highly detrimental to both humans and animals. Most existing vaccines are live attenuated vaccines with safety flaws for people and animals. Therefore, it is advantageous to design a multi-epitope subunit vaccine (MEV) to prevent Brucella infection. To this end, we applied a reverse vaccinology approach. Six cytotoxic T cell (CTL) epitopes, seven T helper cell (HTL) epitopes, and four linear B cell epitopes from CU/ZN-SOD, Omp31, and BP26 were obtained. We linked the CTL, HTL, B-cell epitopes, the appropriate CTB molecular adjuvant, and the universal T helper lymphocyte epitope, PADRE, with linkers AAY, GPPGG, and KK, respectively. This yielded a 412-amino acid MEV construct, which we named MEVcob. The immunogenicity, stability, safety, and feasibility of the construct were evaluated by bioinformatics tools (including the AlphaFold2 prediction tool, the AlphaFold2 tool, NetMHC-I pan 4.0 server, IEDB MHC-I server, ABCpred service, and C-ImmSim server); the physicochemical properties, secondary and tertiary structures, and binding ability of MEVocb to toll-like receptor 4 (TLR4) was analyzed. Then, codon adaptation and computer cloning studies were performed. MEVocb is highly immunogenic in immunostimulation experiments, The proteins translated by these sequences were relatively stable, exhibiting a high antigenic index. Furthermore, mouse experiments confirmed that the MEVocb construct could raise IFN-γ, IgG, IgG2a, IgG1, IL-2, TNF-α levels in mice, indicating that induced a specific humoral and cellular immune response in BALB/c mice. This vaccine induced a statistically significant level of protection in BALB/c mice when challenged with Brucella melitensis 043 in Xinjiang. Briefly, we utilized immunoinformatic tools to design a novel multi-epitope subunit candidate vaccine against Brucella. This vaccine aims to induce host immune responses and confer specific protective effects. The study results offer a theoretical foundation for the development of a novel Brucella subunit vaccine.
Asunto(s)
Vacuna contra la Brucelosis , Brucella melitensis , Brucelosis , Humanos , Animales , Ratones , Ratones Endogámicos BALB C , Proteínas de la Membrana Bacteriana Externa , Brucelosis/prevención & control , Epítopos de Linfocito B , Vacunas de Subunidad , Superóxido Dismutasa , Epítopos de Linfocito T , Biología Computacional/métodos , Simulación del Acoplamiento MolecularRESUMEN
INTRODUCTION: Vaccine formulation with appropriate adjuvants is an attractive approach to develop protective immunity against pathogens. Calcium phosphate nanoparticles (CaPNs) are considered as ideal adjuvants and delivery systems because of their great potential for enhancing immune responses. In the current study, we have designed nanoparticle-based vaccine candidates to induce immune responses and protection against B. melitensis and B. abortus. MATERIALS AND METHODS: For this purpose, we used three Brucella antigens (FliC, 7α-HSDH, BhuA) and two multi-epitopes (poly B and poly T) absorbed by CaPNs. The efficacy of each formulation was evaluated by measuring humoral, cellular and protective responses in immunized mice. RESULTS: The CaPNs showed an average size of about 90 nm with spherical shape and smooth surface. The CaPNs-adsorbed proteins displayed significant increase in cellular and humoral immune responses compared to the control groups. In addition, our results showed increased ratio of specific IgG2a (associated with Th1) to specific IgG1 (associated with Th2). Also, immunized mice with different vaccine candidate formulations were protected against B. melitensis 16M and B. abortus 544, and showed same levels of protection as commercial vaccines (B. melitensis Rev.1 and B. abortus RB51) except for BhuA-CaPNs. DISCUSSION: Our data support the hypothesis that these antigens absorbed with CaPNs could be effective vaccine candidates against B. melitensis and B. abortus.
Asunto(s)
Antígenos Bacterianos/química , Vacuna contra la Brucelosis/química , Vacuna contra la Brucelosis/inmunología , Nanopartículas/química , Adyuvantes Inmunológicos/administración & dosificación , Adyuvantes Inmunológicos/farmacología , Animales , Proteínas Bacterianas/inmunología , Brucella abortus/inmunología , Brucella melitensis/inmunología , Brucelosis/inmunología , Brucelosis/prevención & control , Fosfatos de Calcio/química , Sistemas de Liberación de Medicamentos , Femenino , Inmunidad Humoral , Inmunización , Inmunoglobulina G/inmunología , Proteínas de Transporte de Membrana/inmunología , Ratones Endogámicos BALB CRESUMEN
The intracellular nature of Brucella leads to rise in oxidative stress due to bacterial invasion, particularly at the site of predilection spleen and lymph nodes. The present study aimed to evaluate the erythrocytic and tissue specific oxidative stress responses induced during oil adjuvant killed Brucella melitensis vaccination. The results of the study clearly implicated a significant increase in level of catalase, and superoxide dismutase (SOD) activity and lipid peroxidation (LPO), and total protein content in erythrocytes after vaccination. The activity of glutathione-S-transferase (GST) was unaltered during the period of experiment. The catalase activity and GSH content was significantly increased in lung and spleen tissues. The tissues GST levels increased significantly in all tissues, while tissue SOD level increased significantly only in lung tissues. Thus, it can be inferred that oil adjuvant based Brucella vaccine induces negligible signs of inflammatory pathophysiology and supports the development of significant level of protection against virulent Brucella challenge.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Antioxidantes/metabolismo , Vacuna contra la Brucelosis/administración & dosificación , Brucelosis/prevención & control , Estrés Oxidativo , Animales , Biomarcadores , Vacuna contra la Brucelosis/inmunología , Brucella melitensis , Catalasa/análisis , Eritrocitos/metabolismo , Femenino , Peroxidación de Lípido , Lípidos/administración & dosificación , Ratones , Ratones Endogámicos BALB C , Superóxido Dismutasa/análisis , Vacunas de Productos Inactivados/administración & dosificación , Vacunas de Productos Inactivados/inmunologíaRESUMEN
Brucellosis is an infectious disease that brings enormous economic burdens for developing countries. The Brucella melitensis (B. melitensis) M5-90 vaccine strain (M5-90) has been used on a large scale in China, but may cause abortions if given to pregnant goats or sheep subcutaneously during the late stages of gestation. Moreover, the vaccine M5-90 cannot differentiate natural from vaccinated infection. Therefore, a safer and more potent M5-90 vaccine is required. In this study, a vjbR mutant of M5-90 (M5-90ΔvjbR) was constructed and overcame these drawbacks. M5-90ΔvjbR strain showed reduced survival capability in murine macrophages (RAW 264.7) and BALB/c mice and induced high protective immunity in mice. In addition, M5-90ΔvjbR induced an anti-Brucella-specific immunoglobulin G (IgG) response and stimulated the expression of gamma interferon (INF-γ) and interleukin-4 (IL-4) in vaccinated mice. Furthermore, M5-90ΔvjbR induced IgG response and stimulated the secretion of IFN-γ and IL-4 in immunized sheep. Moreover, the VjbR antigen allowed serological differentiation between infected and vaccinated animals. These results suggest that M5-90ΔvjbR is an ideal live attenuated and efficacious live vaccine candidate against B. melitensis 16â¯M infection.
Asunto(s)
Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Vacuna contra la Brucelosis/inmunología , Brucella melitensis/inmunología , Brucelosis/prevención & control , Modelos Animales de Enfermedad , Animales , Anticuerpos Antibacterianos/inmunología , Proteínas Bacterianas/administración & dosificación , Vacuna contra la Brucelosis/administración & dosificación , Vacuna contra la Brucelosis/genética , Brucella melitensis/genética , Brucelosis/inmunología , Brucelosis/microbiología , Evaluación Preclínica de Medicamentos , Femenino , Eliminación de Gen , Humanos , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-4/genética , Interleucina-4/inmunología , Ratones , Ratones Endogámicos BALB C , Eliminación de Secuencia , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunologíaRESUMEN
We previously developed a potent candidate vaccine against bovine brucellosis caused by Brucella abortus using the influenza viral vector expressing Brucella Omp16 and L7/L12 proteins (Flu-BA). Our success in the Flu-BA vaccine trial in cattle and results of a pilot study in non-pregnant small ruminants prompted us in the current study to test its efficacy against B. melitensis infection in pregnant sheep and goats. In this study, we improved the Flu-BA vaccine formulation and immunization method to achieve maximum efficacy and safety. The Flu-BA vaccine formulation had two additional proteins Omp19 and SOD, and administered thrice with 20% Montanide Gel01 adjuvant, simultaneously by both subcutaneous and conjunctival routes at 21 days intervals in pregnant sheep and goats. At 42 days post-vaccination (DPV) we detected antigen-specific IgG antibodies predominantly of IgG2a isotype but also IgG1, and also detected a strong lymphocyte recall response with IFN-γ production. Importantly, our candidate vaccine prevented abortion in 66.7% and 77.8% of pregnant sheep and goats, respectively. Furthermore, complete protection (absence of live B. melitensis 16M) was observed in 55.6% and 66.7% of challenged sheep and goats, and 72.7% and 90.0% of their fetuses (lambs/yeanlings), respectively. The severity of B. melitensis 16M infection in vaccinated sheep and goats and their fetuses (index of infection and rates of Brucella colonization in tissues) was significantly lower than in control groups. None of the protection parameters after vaccination with Flu-BA vaccine were statistically inferior to protection seen with the commercial B. melitensis Rev.1 vaccine (protection against abortion and vaccination efficacy, alpha = 0.18-0.34, infection index, P = 0.37-0.77, Brucella colonization, P = 0.16 to P > 0.99). In conclusion, our improved Flu-BA vaccine formulation and delivery method were found safe and effective in protecting pregnant sheep and goats against adverse consequences of B. melitensis infection.
Asunto(s)
Linfocitos B/inmunología , Vacuna contra la Brucelosis/inmunología , Brucella melitensis/genética , Brucelosis/prevención & control , Orthomyxoviridae/genética , Linfocitos T/inmunología , Aborto Espontáneo/prevención & control , Animales , Anticuerpos/inmunología , Antígenos Bacterianos/genética , Antígenos Bacterianos/inmunología , Antígenos Bacterianos/metabolismo , Linfocitos B/citología , Linfocitos B/metabolismo , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/inmunología , Proteínas de la Membrana Bacteriana Externa/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Proteínas Bacterianas/metabolismo , Vacuna contra la Brucelosis/genética , Vacuna contra la Brucelosis/metabolismo , Brucella melitensis/patogenicidad , Brucelosis/inmunología , Femenino , Cabras , Hemaglutininas Virales/inmunología , Inmunoglobulina G/inmunología , Interferón gamma/metabolismo , Lipoproteínas/genética , Lipoproteínas/inmunología , Lipoproteínas/metabolismo , Embarazo , Ovinos , Superóxido Dismutasa-1/genética , Superóxido Dismutasa-1/inmunología , Superóxido Dismutasa-1/metabolismo , Linfocitos T/citología , Linfocitos T/metabolismo , VacunaciónRESUMEN
This study was conducted to evaluate the immunogenicity and protective efficacy of a DNA vaccine encoding Brucella abortus Cu,Zn superoxide dismutase (SOD) using the Toll-like receptor 2/6 agonist S-[2,3-bispalmitoyiloxy-(2R)-propyl]-R-cysteinyl-amido-monomethoxy polyethylene glycol (BPPcysMPEG) as an adjuvant. Intranasal coadministration of BPPcysMPEG with a plasmid carrying the SOD-encoding gene (pcDNA-SOD) into BALB/c mice elicited antigen-specific humoral and cellular immune responses. Humoral responses were characterized by the stimulation of IgG2a and IgG1 and by the presence of SOD-specific secretory IgA in nasal and bronchoalveolar lavage fluids. Furthermore, T-cell proliferative responses and increased production of gamma interferon were also observed upon splenocyte restimulation with recombinant SOD. Cytotoxic responses were also stimulated, as demonstrated by the lysis of RB51-SOD-infected J774.A1 macrophages by cells recovered from immunized mice. The pcDNA-SOD/BPPcysMPEG formulation induced improved protection against challenge with the virulent strain B. abortus 2308 in BALB/c mice over that provided by pcDNA-SOD, suggesting the potential of this vaccination strategy against Brucella infection.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Vacuna contra la Brucelosis/inmunología , Brucella abortus/enzimología , Brucelosis/prevención & control , Polietilenglicoles/administración & dosificación , Superóxido Dismutasa/inmunología , Vacunas de ADN/inmunología , Adyuvantes Inmunológicos/farmacología , Administración Intranasal , Animales , Anticuerpos Antibacterianos/sangre , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Líquido del Lavado Bronquioalveolar/inmunología , Vacuna contra la Brucelosis/administración & dosificación , Vacuna contra la Brucelosis/genética , Brucella abortus/genética , Brucelosis/inmunología , Proliferación Celular , Pruebas Inmunológicas de Citotoxicidad , Modelos Animales de Enfermedad , Femenino , Inmunoglobulina A Secretora/análisis , Inmunoglobulina G/sangre , Interferón gamma/metabolismo , Macrófagos/inmunología , Macrófagos/microbiología , Ratones Endogámicos BALB C , Mucosa Nasal/inmunología , Polietilenglicoles/farmacología , Bazo/inmunología , Superóxido Dismutasa/genética , Linfocitos T/inmunología , Receptor Toll-Like 2/agonistas , Receptor Toll-Like 6/agonistas , Vacunas de ADN/administración & dosificación , Vacunas de ADN/genéticaRESUMEN
INTRODUCTION: Brucella melitensis is a facultative intracellular Gram-negative bacterial pathogen that may enter the host via ingestion or inhalation, or through conjunctiva or skin abrasions. Some Brucella spp surface proteins (SPs) play an important role in bacterial adhesion and invasion and thus represent targets for the host immune system. Brucella spp surface protein with apparent molecular mass of 41 kDa interacts selectively with HeLa cells. METHODOLOGY: To evaluate the role of SP41 (41 kDa) as a DNA vaccine against Brucella spp., pCISP41, a plasmid construct for protein expression in mammalian cells, was established. Exogenous SP41 was detected in pCISP41-transfected Vero cell line by immune blotting using specific polyclonal antibody. The protective role of pCISP41 against B. melitensis 16M in mice was evaluated by measuring B and T cell responses in comparison to those achieved with attenuated B. melitensis Rev. 1 vaccine. RESULTS: BALB/c mice injected with pCISP41 were able to develop SP41-specific serum immunoglobulin G (IgG) antibodies. In addition, splenocytes from DNA-SP41-vaccinated mice elicited a T-cell-proliferative response and also induced gamma interferon (IFN-γ) production, but not interleukin-5 (IL-5), suggesting the induction of a T-helper-1-dominated immune response. Vaccination with attenuated B. melitensis Rev.1 strain induced better protection levels than DNA vaccination with SP41 against B. melitensis 16M in mice. CONCLUSIONS: Such responses play an important role against intracellular infecting agents such as Brucella spp. Altogether, our data suggest that SP41 may represent a promising candidate for DNA vaccination against brucellosis, but more investigation to increase its protective efficacy should be done.
Asunto(s)
Vacuna contra la Brucelosis/inmunología , Brucella melitensis/inmunología , Brucelosis/prevención & control , Vacunas de ADN/inmunología , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Vacuna contra la Brucelosis/administración & dosificación , Brucelosis/inmunología , Chlorocebus aethiops , Evaluación Preclínica de Medicamentos , Humanos , Inmunidad Celular , Inmunidad Humoral , Interferón gamma/inmunología , Interleucina-5/inmunología , Ratones , Ratones Endogámicos BALB C , Plásmidos/genética , Plásmidos/metabolismo , Vacunas de ADN/administración & dosificación , Células VeroRESUMEN
Due to drawbacks of live attenuated vaccines, much more attention has been focused on screening of Brucella protective antigens as subunit vaccine candidates. Brucella is a facultative intracellular bacterium and cell mediated immunity plays essential roles for protection against Brucella infection. Identification of Brucella antigens that present T-cell epitopes to the host could enable development of such vaccines. In this study, 45 proven or putative pathogenesis-associated factors of Brucella were selected according to currently available data. After expressed and purified, 35 proteins were qualified for analysis of their abilities to stimulate T-cell responses in vitro. Then, an in vitro gamma interferon (IFN-γ) assay was used to identify potential T-cell antigens from B. abortus. In total, 7 individual proteins that stimulated strong IFN-γ responses in splenocytes from mice immunized with B. abortus live vaccine S19 were identified. The protective efficiencies of these 7 recombinant proteins were further evaluated. Mice given BAB1_1316 (CobB) or BAB1_1688 (AsnC) plus adjuvant could provide protection against virulent B. abortus infection, similarly with the known protective antigen Cu-Zn SOD and the license vaccine S19. In addition, CobB and AsnC could induce strong antibodies responses in BALB/c mice. Altogether, the present study showed that CobB or AsnC protein could be useful antigen candidates for the development of subunit vaccines against brucellosis with adequate immunogenicity and protection efficacy.
Asunto(s)
Brucella abortus/metabolismo , Brucelosis/prevención & control , Proteínas de Escherichia coli/metabolismo , Sirtuinas/metabolismo , Transactivadores/metabolismo , Animales , Antígenos Bacterianos/inmunología , Vacuna contra la Brucelosis/inmunología , Brucelosis/inmunología , Femenino , Sistema Inmunológico , Inmunización , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/metabolismo , Bazo/citología , Superóxido Dismutasa/metabolismo , Linfocitos T/metabolismo , Linfocitos T/microbiologíaRESUMEN
Brucellosis is a worldwide zoonotic disease caused by Brucella abortus and a number of closely related species. Brucellosis has severe impact on the health and economic prosperity of the developing countries due to the persistent nature of infection and unavailability of effective control measures. The Cu-Zn superoxide dismuatse (SOD) protein of Brucella have been extensively studied as a major antigen involved in bacterial evading mechanism of host defence. Being a critical pro-inflammatory cytokine interleukin-18 (IL-18) plays key role in induction of immune mediated protection against intracellular pathogens. In the present study, we aimed to investigate the immunogenic potential of fusogenic liposomes (escheriosomes) encapsulated recombinant Cu-Zn SOD (rSOD) protein alone or in combination with recombinant IL-18 (rIL-18). Escheriosomes encapsulated rSOD mediated immune responses were further increased upon co-immunization with rIL-18. Furthermore, immunization with escheriosomes encapsulated rSOD alone or in combination with rIL-18, increased resistance in mice against challenge with B. abortus 544.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Vacuna contra la Brucelosis/inmunología , Brucella abortus/inmunología , Inmunización/métodos , Interleucina-18/administración & dosificación , Liposomas/administración & dosificación , Superóxido Dismutasa/inmunología , Animales , Antígenos Bacterianos/genética , Antígenos Bacterianos/inmunología , Vacuna contra la Brucelosis/administración & dosificación , Vacuna contra la Brucelosis/genética , Brucella abortus/genética , Brucelosis/prevención & control , Modelos Animales de Enfermedad , Femenino , Ratones , Enfermedades de los Roedores/prevención & control , Superóxido Dismutasa/genética , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunologíaRESUMEN
The water buffalo is an important domestic animal worldwide, and the local Buffalypso variety was developed in Trinidad to have improved beef qualities. Brucellosis was diagnosed in Trinidad and Tobago during 1998 in both cattle and domestic water buffalo (Bubalus bubalis) populations. Brucellosis in the latter species is caused by infection with Brucella abortus, similar to bovine brucellosis. Control of brucellosis is of paramount importance to preservation of the genetic diversity of these animals in Trinidad, and this has been complicated by differences in the epidemiology of water buffalo and bovine brucellosis. Some diagnostic tests do not have comparable accuracy between the two species, and the RB51 vaccine does not adequately protect against infection in water buffalo. The water buffalo in Trinidad may also be more resistant to infection than cattle. Development of effective vaccination protocols is key to brucellosis control in Buffalypso in Trinidad, and prohibitions on import of virulent B. abortus strains for vaccine efficacy studies has impeded progress in this area. These Trinidadian strains are of variable virulence; some might be effective for challenge in vaccine efficacy studies, while other, of lower virulence, may be vaccine candidates for use in water buffalo.
Asunto(s)
Vacuna contra la Brucelosis/uso terapéutico , Brucella abortus/inmunología , Brucella abortus/patogenicidad , Brucelosis Bovina/epidemiología , Brucelosis/veterinaria , Búfalos/microbiología , Animales , Brucelosis/diagnóstico , Brucelosis/epidemiología , Brucelosis/microbiología , Brucelosis Bovina/diagnóstico , Brucelosis Bovina/microbiología , Bovinos , Prevalencia , Trinidad y Tobago/epidemiología , Vacunación/veterinaria , VirulenciaRESUMEN
Use of antibiotics can't completely solve the problem of brucellosis treatment, especially its chronic forms, because antibacterial preparations do not eliminate main pathogenetic factor of the disease--sensibilization of the macroorganism. It makes actual the question about complex immuno- and antibacterial therapy. Long-term clinical experience proved high effectiveness of a therapeutic brucellosis vaccine. Earlier this preparation was manufactured in Research Institute of Vaccines and Sera in Tbilisi (Georgia). To date new composition of components of the vaccine has been developed, and manufacturing and control methods have been improved. Marked desensitizing effect of the vaccine and its stimulatory action on cellular and humoral immunity has been observed. In 2002 technological normative documentation for manufacturing and use of the vaccine was developed in the Research Institute of Microbiology (Kirov) and production of the vaccine began.
Asunto(s)
Vacuna contra la Brucelosis/uso terapéutico , Brucella abortus/inmunología , Brucella melitensis/inmunología , Brucelosis/terapia , Aglutininas/sangre , Animales , Vacuna contra la Brucelosis/administración & dosificación , Vacuna contra la Brucelosis/aislamiento & purificación , Vacuna contra la Brucelosis/normas , Brucella abortus/crecimiento & desarrollo , Brucella melitensis/crecimiento & desarrollo , Brucelosis/sangre , Brucelosis/inmunología , Criopreservación , Medios de Cultivo , Evaluación Preclínica de Medicamentos , Femenino , Cobayas , Inyecciones Subcutáneas , Masculino , FagocitosisRESUMEN
An experiment was conducted to investigate the effects of the lectin, Concanavalin A (Con A), contained in raw Jack bean (JB) (Canavalia ensiformis, L.) seeds on the immunological response of broilers. A maize-soybean meal basal diet was prepared to which either 2.5, 5, or 10% of ground raw Jack bean (RJB) seeds was added. The RJB seeds contained 24 g Con A/kg on a dry matter basis, as measured by rocket immunoelectrophoresis. Similar diets were prepared by using the same levels of JB after toasting at 190 C for 16 min. In addition, the basal diet was pair-fed to groups of chicks at the level of feed intake of chicks fed the 10% RJB diet. Each diet was fed to six groups of six chicks for 6 wk. At 5 wk, 15 of chicks from each diet were immunized against Brucella abortus (BA) and the anti-BA antibody titers were determined 1 wk later by ELISA. Antibody production against Con A was also measured by the same method. Binding of Con A to intestinal villi and subsequent endocytosis were confirmed by microscopic examination using a specific peroxidase-antiperoxidase-staining technique. Performance was recorded weekly. Feed intake and weight gain were reduced (P < 0.05) only by the diet containing 10% RJB, indicating that broiler chicks can tolerate daily intakes of 100 mg of Con A over 6 wk without affecting growth. Toasted JB diets supported adequate chick performance. The antibody response to BA did not differ with dietary treatment. Serum from chicks fed raw JB also contained antibodies against Con A. The bursa of Fabricius, thymus, spleen, and pancreas dry weights, as a percentage of dry body weight, were not affected by the experimental diets. The data indicated that Con A binds to the cells of the gastrointestinal tract, passes into the general circulation and, eventually, elicits an immunological response without affecting the production of antibodies to BA.
Asunto(s)
Anticuerpos Antibacterianos/biosíntesis , Peso Corporal/efectos de los fármacos , Pollos/inmunología , Concanavalina A/farmacología , Duodeno/efectos de los fármacos , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Peso Corporal/fisiología , Vacuna contra la Brucelosis/inmunología , Brucella abortus/inmunología , Pollos/fisiología , Estudios de Cohortes , Concanavalina A/administración & dosificación , Concanavalina A/farmacocinética , Duodeno/metabolismo , Duodeno/ultraestructura , Ingestión de Alimentos/fisiología , Fabaceae/química , Fabaceae/inmunología , Sueros Inmunes/inmunología , Inmunoglobulina G/inmunología , Hígado/anatomía & histología , Hígado/efectos de los fármacos , Masculino , Microvellosidades/efectos de los fármacos , Microvellosidades/ultraestructura , Tamaño de los Órganos/efectos de los fármacos , Lectinas de Plantas , Plantas Medicinales , Conejos , Distribución Aleatoria , Semillas/química , Semillas/inmunologíaRESUMEN
Neonatal lambs and adult wethers were passively immunized with ovine antibody directed against ovalbumin or Brucella abortus. Estimates for the biological half-lives of the antibodies ranged from 18 to 24 days in neonatal lambs and 12 to 17 days in adult wethers. The evidence suggested that both normal and immunosuppressed wethers which were passively immunized with serum antibody catabolized this antibody at a faster rate than did neonatal lambs. The data provided no support for the hypothesis that the growth factors and immunomodulatory factors, which are known to be present in colostrum, can influence the biological half-life of homologous antibody following passive immunization.
Asunto(s)
Animales Recién Nacidos/inmunología , Brucella abortus/inmunología , Inmunización Pasiva , Ovalbúmina/inmunología , Ovinos/inmunología , Animales , Anticuerpos Antibacterianos/inmunología , Vacuna contra la Brucelosis/administración & dosificación , Calostro/inmunología , Femenino , Semivida , Ovalbúmina/administración & dosificación , Embarazo , Análisis de RegresiónRESUMEN
Rams which were vaccinated at 6-8 months of age with a water-in-oil Brucella ovis-vitamin E adjuvant vaccine had significantly higher serum antibody levels than rams vaccinated with a commercial B. ovis bacterin or B. melitensis Rev 1. The adjuvant vaccine did not cause abscesses at the site of injection as some water-in-oil emulsions do. Two years after vaccination, the vitamin E adjuvant-vaccinated rams had higher antibody level than the other groups. This was most likely due to a secondary response to naturally occurring infection with B. ovis.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Vacuna contra la Brucelosis/inmunología , Ovinos/inmunología , Vitamina E/farmacología , Animales , Anticuerpos Antibacterianos/biosíntesis , Brucella/inmunología , Brucelosis/prevención & control , Brucelosis/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Epididimitis/prevención & control , Epididimitis/veterinaria , Masculino , Enfermedades de las Ovejas/prevención & controlRESUMEN
Twenty-four 7-mo-old beef heifers (Charolais Simmental cross), weighing 213 kg, were used to determine the effect of vitamin E (VitE) and(or) selenium (Se) supplementation on the humoral response to a standard dose of Brucella abortus strain 19 vaccine and on the levels of naturally occurring immunoglobulins (Ig) to several antigens. The treatments were as follows: Group 1, no supplement; Group 2, supplementation with 6 g of elemental Se; Group 3, supplementation with 1,400 IU/d of VitE; and Group 4, Se and VitE supplements combined. There were no significant differences in anti-B. abortus IgG1, IgG2, or IgM antibody levels due to Se, VitE or Se/VitE treatments; the concentrations of IgA antibody were too low to be measured with the ELISA test used. Statistical analysis revealed that the levels of total and IgM natural antibody to Salmonella typhimurium were higher in Group 3. Perhaps VitE supplementation given in conjunction with B. abortus vaccine enhanced the production of antibody to S. typhimurium in several animals whose humoral system had been activated by previous exposure to this organism.
Asunto(s)
Anticuerpos Antibacterianos/biosíntesis , Vacuna contra la Brucelosis/inmunología , Brucelosis Bovina/inmunología , Selenio/farmacología , Vitamina E/farmacología , Animales , Brucella abortus/inmunología , Bovinos , Femenino , Inmunoglobulinas/biosíntesis , Lipopolisacáridos/inmunología , Distribución Aleatoria , Salmonella typhimurium/inmunología , Selenio/sangre , Vacunación/veterinaria , Vitamina E/sangreRESUMEN
The possibility of evaluating the immunogenic potency of brucellosis vaccine BA-19 by immunological methods has been shown. Morphometry and the quantitative evaluation of globulin-producing cells in lymphoid organs by direct and indirect immunofluorescent techniques serve as informative evaluation tests. The marker method with the evaluation of lymphocyte classes by acid and alkaline phosphatases and the electrophoretic motility characteristics of T- and B-lymphocytes, used in combination with the above-mentioned methods, present information on immunogenesis. The immunoperoxidase method and the values of the opsonophagocytic index characterize the state of phagocytosis and the persistence of the antigen.
Asunto(s)
Vacuna contra la Brucelosis/inmunología , Animales , Linfocitos B/citología , Linfocitos B/inmunología , Vacuna contra la Brucelosis/administración & dosificación , Brucella abortus/inmunología , Citofotometría , Evaluación Preclínica de Medicamentos , Cobayas , Inmunoglobulinas/biosíntesis , Ganglios Linfáticos/anatomía & histología , Ganglios Linfáticos/inmunología , Fagocitosis , Bazo/anatomía & histología , Bazo/inmunología , Linfocitos T/citología , Linfocitos T/inmunología , Factores de TiempoAsunto(s)
Antígenos Bacterianos/aislamiento & purificación , Vacuna contra la Brucelosis/aislamiento & purificación , Brucella abortus/inmunología , Animales , Antígenos Bacterianos/inmunología , Antígenos Bacterianos/normas , Vacuna contra la Brucelosis/inmunología , Vacuna contra la Brucelosis/normas , Brucelosis/prevención & control , Fenómenos Químicos , Química Física , Perros , Evaluación Preclínica de Medicamentos , Cobayas , Humanos , Macaca fascicularis , Ratones , Fenoles , Control de Calidad , SolubilidadRESUMEN
The immune response of cattle to nonliving vaccines derived from Brucella abortus rough strain 45/20 was studied. Vaccines contained trehalose dimycolate and a derivative of muramyl dipeptide. N-acetylmuramyl-L-alpha-aminobutyryl-D-isoglutamine. A factorial experiment was designed to test the effects of type of antigen, quantity of antigen, and quantity of mineral oil on the immune response to porin. Muramyl dipeptide was kept constant at 5 mg per dose, and 1 part of trehalose dimycolate was incorporated for two parts of dry matter. Over a 10-week period, blastogenesis responses to porin were largest in cattle immunized with outer membranes; the highest antibody titers to the porin-lipopolysaccharide complex were achieved by immunization with detergent-extracted outer membrane proteins. There was no advantage in the use of 25, rather than 5, mg of any of the antigens, but antibody responses were improved by increasing the quantity of oil from 0.6 to 1.8 ml per dose. In other animals, blastogenesis and antibody responses were sustained at high levels longer than 3 months after two vaccinations with outer membrane proteins. Intradermal injection of porin evoked inflammatory reactions histologically consistent with delayed-type hypersensitivity. Cross-reactions in cases of delayed-type hypersensitivity occurred with porin derived from a smooth strain of B. abortus but were less extensive than in the blastogenesis test. The magnitude of the delayed-type hypersensitivity and blastogenesis responses induced by vaccination exceeded those observed after natural or experimental infections. No ill effects were observed after vaccination. These findings provide a basis for the use of trehalose dimycolate and muramyl dipeptide adjuvants in evaluating nonviable vaccines for bovine brucellosis.
Asunto(s)
Acetilmuramil-Alanil-Isoglutamina/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Brucella abortus/inmunología , Factores Cordón/inmunología , Glucolípidos/inmunología , Proteínas de la Membrana/inmunología , Animales , Proteínas de la Membrana Bacteriana Externa , Vacuna contra la Brucelosis/inmunología , Brucelosis Bovina/inmunología , Brucelosis Bovina/prevención & control , Bovinos , Relación Dosis-Respuesta Inmunológica , Evaluación Preclínica de Medicamentos/veterinaria , Inmunización/veterinaria , PorinasRESUMEN
Guinea pigs were subcutaneously inoculated with 300 microgram of Brucella abortus strain 45/20 killed cells combined in 1% oil emulsion with trehalose dimycolate (TDM), muramyl dipeptide (MDP), or a combination of the 2 immunopotentiators. Protection, as determined by splenic infections in the guinea pigs after challenge exposure, was compared with that induced by strain 19 vaccine. With few exceptions, protection induced by bacterins containing 50 to 1,000 microgram of TDM or TDM-MDP/dose was comparable with that of strain 19 vaccine (P greater than 0.05). Bacterins that contained MDP as an adjuvant were inferior to those with TDM regardless of the excipient or method of preparation. There was no further enhancement of immunogenicity by the addition of MDP to bacterins that already contained TDM. Mineral oil could not be replaced by a metabolizable excipient in bacterins potentiated with TDM.
Asunto(s)
Acetilmuramil-Alanil-Isoglutamina/inmunología , Adyuvantes Inmunológicos , Vacuna contra la Brucelosis , Brucella abortus/inmunología , Brucelosis/veterinaria , Factores Cordón/inmunología , Glucolípidos/inmunología , Glicopéptidos/inmunología , Animales , Brucelosis/prevención & control , CobayasRESUMEN
A cell wall fraction (F8) extracted by boiling sodium dodecylsulfate at 4 % from Brucella abortus 99S was used with oil adjuvant to vaccinate groups of ten guinea-pigs, at doses equivalent to 1 X 10(9) and 1 X 10(10) bacteria, once or twice at 3 month intervals. H38 vaccine, a total cell vaccine from formalized B. melitensis 53 H38, was used as a reference, at doses 3 X 10(8) and 3 X 10(9) bacteria. These doses were chosen since they have about the same vaccinal activity in mice being respectively equal to 10 and 100 mice optimal dose (MOD). One extra-group of guinea-pigs received two injections of 100 microgram of smooth-lipopolysaccharide (LPS-S) of B. melitensis 16M, in adjuvant. Control group received the adjuvant only. Guinea-pigs were challenged 3 months after the last vaccination with 5,000 colony-forming units of B. abortus 544, and autopsied 40 days later. The spleen and 8 lymph nodes were cultured: a guinea-pig is considered as protected if no Brucella was found in any sample. Protection afforded by the two vaccines is dose-dependent. H38 vaccine gives a better protection (infected 24 %) than F8 (46 %) since a higher dose is needed to obtain the same level of protection: i. e., 100 MOD of F8 is about equal to 10 MOD of H38 (35 and 37 % respectively). Contrary to what was previously shown in mice, recall does not improve the immunity and LPS-S does not vaccinate at all.