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1.
Vaccine ; 32(24): 2794-8, 2014 May 19.
Article in English | MEDLINE | ID: mdl-24593996

ABSTRACT

Rotavirus is the most common cause of severe diarrhea in many animal species of economic interest. A simple, safe and cost-effective vaccine is required for the control and prevention of rotavirus in animals. In this study, we evaluated the use of Saccharomyces cerevisiae extracts containing rotavirus-like particles (RLP) as a vaccine candidate in an adult mice model. Two doses of 1mg of yeast extract containing rotavirus proteins (between 0.3 and 3 µg) resulted in an immunological response capable of reducing the replication of rotavirus after infection. Viral shedding in all mice groups diminished in comparison with the control group when challenged with 100 50% diarrhea doses (DD50) of murine rotavirus strain EDIM. Interestingly, when immunizing intranasally protection against rotavirus infection was observed even when no increase in rotavirus-specific antibody titers was evident, suggesting that cellular responses were responsible of protection. Our results indicate that raw yeast extracts containing rotavirus proteins and RLP are a simple, cost-effective alternative for veterinary vaccines against rotavirus.


Subject(s)
Rotavirus Infections/prevention & control , Rotavirus Vaccines/immunology , Saccharomyces cerevisiae/virology , Vaccines, Virus-Like Particle/immunology , Animals , Antibodies, Viral/blood , Antibody Formation , Antigens, Viral/immunology , Batch Cell Culture Techniques , Capsid Proteins/immunology , Female , Mice , Rotavirus , Saccharomyces cerevisiae/immunology , Virus Cultivation , Virus Shedding
2.
Virol J ; 6: 17, 2009 Feb 06.
Article in English | MEDLINE | ID: mdl-19196485

ABSTRACT

BACKGROUND: Gag protein from HIV-1 is a polyprotein of 55 kDa, which, during viral maturation, is cleaved to release matrix p17, core p24 and nucleocapsid proteins. The p24 antigen contains epitopes that prime helper CD4 T-cells, which have been demonstrated to be protective and it can elicit lymphocyte proliferation. Thus, p24 is likely to be an integral part of any multicomponent HIV vaccine. The availability of an optimal adjuvant and carrier to enhance antiviral responses may accelerate the development of a vaccine candidate against HIV. The aim of this study was to investigate the adjuvant-carrier properties of the B ricin subunit (RTB) when fused to p24. RESULTS: A fusion between ricin toxin B subunit and p24 HIV (RTB/p24) was expressed in E. coli. Affinity chromatography was used for purification of p24 alone and RTB/p24 from cytosolic fractions. Biological activity of RTB/p24 was determined by ELISA and affinity chromatography using the artificial receptor glycoprotein asialofetuin. Both assays have demonstrated that RTB/p24 is able to interact with complex sugars, suggesting that the chimeric protein retains lectin activity. Also, RTB/p24 was demonstrated to be immunologically active in mice. Two weeks after intraperitoneal inoculation with RTB/p24 without an adjuvant, a strong anti-p24 immune response was detected. The levels of the antibodies were comparable to those found in mice immunized with p24 alone in the presence of Freund adjuvant. RTB/p24 inoculated intranasally in mice, also elicited significant immune responses to p24, although the response was not as strong as that obtained in mice immunized with p24 in the presence of the mucosal adjuvant cholera toxin. CONCLUSION: In this work, we report the expression in E. coli of HIV-1 p24 fused to the subunit B of ricin toxin. The high levels of antibodies obtained after intranasal and intraperitoneal immunization of mice demonstrate the adjuvant-carrier properties of RTB when conjugated to an HIV structural protein. This is the first report in which a eukaryotic toxin produced in E. coli is employed as an adjuvant to elicit immune responses to p24 HIV core antigen.


Subject(s)
Escherichia coli/genetics , HIV Core Protein p24/immunology , HIV Core Protein p24/isolation & purification , HIV Infections/immunology , HIV-1/immunology , Ricin/immunology , Ricin/isolation & purification , Adjuvants, Immunologic/genetics , Adjuvants, Immunologic/isolation & purification , Animals , Antibodies, Viral/blood , Escherichia coli/metabolism , Female , Gene Expression , HIV Core Protein p24/genetics , HIV Infections/virology , HIV-1/chemistry , Humans , Immunization , Mice , Mice, Inbred BALB C , Protein Engineering , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/isolation & purification , Ricin/genetics
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