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1.
Microb Pathog ; 124: 250-257, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30149131

RESUMO

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 16 M infection.


Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Vacina contra Brucelose/imunologia , Brucella melitensis/imunologia , Brucelose/prevenção & controle , Modelos Animais de Doenças , Animais , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/administração & dosagem , Vacina contra Brucelose/administração & dosagem , Vacina contra Brucelose/genética , Brucella melitensis/genética , Brucelose/imunologia , Brucelose/microbiologia , Avaliação Pré-Clínica de Medicamentos , Feminino , Deleção de Genes , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Deleção de Sequência , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia
2.
PLoS One ; 12(10): e0186484, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29023541

RESUMO

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.


Assuntos
Linfócitos B/imunologia , Vacina contra Brucelose/imunologia , Brucella melitensis/genética , Brucelose/prevenção & controle , Orthomyxoviridae/genética , Linfócitos T/imunologia , Aborto Espontâneo/prevenção & controle , Animais , Anticorpos/imunologia , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/metabolismo , Linfócitos B/citologia , Linfócitos B/metabolismo , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Vacina contra Brucelose/genética , Vacina contra Brucelose/metabolismo , Brucella melitensis/patogenicidade , Brucelose/imunologia , Feminino , Cabras , Hemaglutininas Virais/imunologia , Imunoglobulina G/imunologia , Interferon gama/metabolismo , Lipoproteínas/genética , Lipoproteínas/imunologia , Lipoproteínas/metabolismo , Gravidez , Ovinos , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/imunologia , Superóxido Dismutase-1/metabolismo , Linfócitos T/citologia , Linfócitos T/metabolismo , Vacinação
3.
Clin Vaccine Immunol ; 21(11): 1474-80, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25165025

RESUMO

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.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Vacina contra Brucelose/imunologia , Brucella abortus/enzimologia , Brucelose/prevenção & controle , Polietilenoglicóis/administração & dosagem , Superóxido Dismutase/imunologia , Vacinas de DNA/imunologia , Adjuvantes Imunológicos/farmacologia , Administração Intranasal , Animais , Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Vacina contra Brucelose/administração & dosagem , Vacina contra Brucelose/genética , Brucella abortus/genética , Brucelose/imunologia , Proliferação de Células , Testes Imunológicos de Citotoxicidade , Modelos Animais de Doenças , Feminino , Imunoglobulina A Secretora/análise , Imunoglobulina G/sangue , Interferon gama/metabolismo , Macrófagos/imunologia , Macrófagos/microbiologia , Camundongos Endogâmicos BALB C , Mucosa Nasal/imunologia , Polietilenoglicóis/farmacologia , Baço/imunologia , Superóxido Dismutase/genética , Linfócitos T/imunologia , Receptor 2 Toll-Like/agonistas , Receptor 6 Toll-Like/agonistas , Vacinas de DNA/administração & dosagem , Vacinas de DNA/genética
4.
Vaccine ; 29(29-30): 4720-7, 2011 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-21565241

RESUMO

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.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Vacina contra Brucelose/imunologia , Brucella abortus/imunologia , Imunização/métodos , Interleucina-18/administração & dosagem , Lipossomos/administração & dosagem , Superóxido Dismutase/imunologia , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Vacina contra Brucelose/administração & dosagem , Vacina contra Brucelose/genética , Brucella abortus/genética , Brucelose/prevenção & controle , Modelos Animais de Doenças , Feminino , Camundongos , Doenças dos Roedores/prevenção & controle , Superóxido Dismutase/genética , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
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