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Immunogenic profiling in mice of a HIV/AIDS vaccine candidate (MVA-B) expressing four HIV-1 antigens and potentiation by specific gene deletions.

García-Arriaza J, Nájera JL, Gómez CE, Sorzano CO, Esteban M - PLoS ONE (2010)

Bottom Line: Both vectors were capable of producing similar levels of antibodies against Env.These findings revealed that MVA-B and MVA-B DeltaA41L/DeltaB16R induced in mice robust, polyfunctional and durable T-cell responses to HIV-1 antigens, but the double deletion mutant showed enhanced magnitude and quality of HIV-1 adaptive and memory responses.Our observations are relevant in the immune evaluation of MVA-B and on improvements of MVA vectors as HIV-1 vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain.

ABSTRACT

Background: The immune parameters of HIV/AIDS vaccine candidates that might be relevant in protection against HIV-1 infection are still undefined. The highly attenuated poxvirus strain MVA is one of the most promising vectors to be use as HIV-1 vaccine. We have previously described a recombinant MVA expressing HIV-1 Env, Gag, Pol and Nef antigens from clade B (referred as MVA-B), that induced HIV-1-specific immune responses in different animal models and gene signatures in human dendritic cells (DCs) with immunoregulatory function.

Methodology/principal findings: In an effort to characterize in more detail the immunogenic profile of MVA-B and to improve its immunogenicity we have generated a new vector lacking two genes (A41L and B16R), known to counteract host immune responses by blocking the action of CC-chemokines and of interleukin 1beta, respectively (referred as MVA-B DeltaA41L/DeltaB16R). A DNA prime/MVA boost immunization protocol was used to compare the adaptive and memory HIV-1 specific immune responses induced in mice by the parental MVA-B and by the double deletion mutant MVA-B DeltaA41L/DeltaB16R. Flow cytometry analysis revealed that both vectors triggered HIV-1-specific CD4(+) and CD8(+) T cells, with the CD8(+) T-cell compartment responsible for >91.9% of the total HIV-1 responses in both immunization groups. However, MVA-B DeltaA41L/DeltaB16R enhanced the magnitude and polyfunctionality of the HIV-1-specific CD4(+) and CD8(+) T-cell immune responses. HIV-1-specific CD4(+) T-cell responses were polyfunctional and preferentially Env-specific in both immunization groups. Significantly, while MVA-B induced preferentially Env-specific CD8(+) T-cell responses, MVA-B DeltaA41L/DeltaB16R induced more GPN-specific CD8(+) T-cell responses, with an enhanced polyfunctional pattern. Both vectors were capable of producing similar levels of antibodies against Env.

Conclusions/significance: These findings revealed that MVA-B and MVA-B DeltaA41L/DeltaB16R induced in mice robust, polyfunctional and durable T-cell responses to HIV-1 antigens, but the double deletion mutant showed enhanced magnitude and quality of HIV-1 adaptive and memory responses. Our observations are relevant in the immune evaluation of MVA-B and on improvements of MVA vectors as HIV-1 vaccines.

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Humoral immune response elicited against HIV-1 gp160 protein induced by immunization with MVA-B and MVA-B ΔA41L/ΔB16R.Serum from individually immunized mice was evaluated by ELISA for specific anti-gp120 antibodies in blood taken 11 (A) and 53 (B) days after the last immunization with DNA-B/MVA-B, DNA-B/MVA-B ΔA41L/ΔB16R and DNA-φ/MVA-WT, as described under Materials and Methods. Serum from naïve (not immunized) animals served as control. Absorbance values (measured at 450 nm) correspond to 1/100 dilution of individual serum, and each mouse is represented by a dot. A black dash line reveals the mean value for each group. A representative experiment out of two is shown.
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pone-0012395-g006: Humoral immune response elicited against HIV-1 gp160 protein induced by immunization with MVA-B and MVA-B ΔA41L/ΔB16R.Serum from individually immunized mice was evaluated by ELISA for specific anti-gp120 antibodies in blood taken 11 (A) and 53 (B) days after the last immunization with DNA-B/MVA-B, DNA-B/MVA-B ΔA41L/ΔB16R and DNA-φ/MVA-WT, as described under Materials and Methods. Serum from naïve (not immunized) animals served as control. Absorbance values (measured at 450 nm) correspond to 1/100 dilution of individual serum, and each mouse is represented by a dot. A black dash line reveals the mean value for each group. A representative experiment out of two is shown.

Mentions: Since all the viral vectors release monomeric gp120 from cells in the course of virus infection [7], we also evaluated whether DNA-B/MVA-B and DNA-B/MVA-B ΔA41L/ΔB16R immunization groups elicited an antibody response against HIV-1 Env. This was performed by ELISA using individual mouse serum from each group of immunized animals at 11 and 53 days post-boost. As shown in Figure 6A and 6B, between both immunization groups, similar levels of specific antibodies reactive against gp160 protein from the HIV-1 clone LAV (clade B) were observed at the different times post-boost. Therefore, both immunization groups induced humoral immune responses against HIV-1 Env and the viral deletions did not affect the antibody levels.


Immunogenic profiling in mice of a HIV/AIDS vaccine candidate (MVA-B) expressing four HIV-1 antigens and potentiation by specific gene deletions.

García-Arriaza J, Nájera JL, Gómez CE, Sorzano CO, Esteban M - PLoS ONE (2010)

Humoral immune response elicited against HIV-1 gp160 protein induced by immunization with MVA-B and MVA-B ΔA41L/ΔB16R.Serum from individually immunized mice was evaluated by ELISA for specific anti-gp120 antibodies in blood taken 11 (A) and 53 (B) days after the last immunization with DNA-B/MVA-B, DNA-B/MVA-B ΔA41L/ΔB16R and DNA-φ/MVA-WT, as described under Materials and Methods. Serum from naïve (not immunized) animals served as control. Absorbance values (measured at 450 nm) correspond to 1/100 dilution of individual serum, and each mouse is represented by a dot. A black dash line reveals the mean value for each group. A representative experiment out of two is shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2927552&req=5

pone-0012395-g006: Humoral immune response elicited against HIV-1 gp160 protein induced by immunization with MVA-B and MVA-B ΔA41L/ΔB16R.Serum from individually immunized mice was evaluated by ELISA for specific anti-gp120 antibodies in blood taken 11 (A) and 53 (B) days after the last immunization with DNA-B/MVA-B, DNA-B/MVA-B ΔA41L/ΔB16R and DNA-φ/MVA-WT, as described under Materials and Methods. Serum from naïve (not immunized) animals served as control. Absorbance values (measured at 450 nm) correspond to 1/100 dilution of individual serum, and each mouse is represented by a dot. A black dash line reveals the mean value for each group. A representative experiment out of two is shown.
Mentions: Since all the viral vectors release monomeric gp120 from cells in the course of virus infection [7], we also evaluated whether DNA-B/MVA-B and DNA-B/MVA-B ΔA41L/ΔB16R immunization groups elicited an antibody response against HIV-1 Env. This was performed by ELISA using individual mouse serum from each group of immunized animals at 11 and 53 days post-boost. As shown in Figure 6A and 6B, between both immunization groups, similar levels of specific antibodies reactive against gp160 protein from the HIV-1 clone LAV (clade B) were observed at the different times post-boost. Therefore, both immunization groups induced humoral immune responses against HIV-1 Env and the viral deletions did not affect the antibody levels.

Bottom Line: Both vectors were capable of producing similar levels of antibodies against Env.These findings revealed that MVA-B and MVA-B DeltaA41L/DeltaB16R induced in mice robust, polyfunctional and durable T-cell responses to HIV-1 antigens, but the double deletion mutant showed enhanced magnitude and quality of HIV-1 adaptive and memory responses.Our observations are relevant in the immune evaluation of MVA-B and on improvements of MVA vectors as HIV-1 vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain.

ABSTRACT

Background: The immune parameters of HIV/AIDS vaccine candidates that might be relevant in protection against HIV-1 infection are still undefined. The highly attenuated poxvirus strain MVA is one of the most promising vectors to be use as HIV-1 vaccine. We have previously described a recombinant MVA expressing HIV-1 Env, Gag, Pol and Nef antigens from clade B (referred as MVA-B), that induced HIV-1-specific immune responses in different animal models and gene signatures in human dendritic cells (DCs) with immunoregulatory function.

Methodology/principal findings: In an effort to characterize in more detail the immunogenic profile of MVA-B and to improve its immunogenicity we have generated a new vector lacking two genes (A41L and B16R), known to counteract host immune responses by blocking the action of CC-chemokines and of interleukin 1beta, respectively (referred as MVA-B DeltaA41L/DeltaB16R). A DNA prime/MVA boost immunization protocol was used to compare the adaptive and memory HIV-1 specific immune responses induced in mice by the parental MVA-B and by the double deletion mutant MVA-B DeltaA41L/DeltaB16R. Flow cytometry analysis revealed that both vectors triggered HIV-1-specific CD4(+) and CD8(+) T cells, with the CD8(+) T-cell compartment responsible for >91.9% of the total HIV-1 responses in both immunization groups. However, MVA-B DeltaA41L/DeltaB16R enhanced the magnitude and polyfunctionality of the HIV-1-specific CD4(+) and CD8(+) T-cell immune responses. HIV-1-specific CD4(+) T-cell responses were polyfunctional and preferentially Env-specific in both immunization groups. Significantly, while MVA-B induced preferentially Env-specific CD8(+) T-cell responses, MVA-B DeltaA41L/DeltaB16R induced more GPN-specific CD8(+) T-cell responses, with an enhanced polyfunctional pattern. Both vectors were capable of producing similar levels of antibodies against Env.

Conclusions/significance: These findings revealed that MVA-B and MVA-B DeltaA41L/DeltaB16R induced in mice robust, polyfunctional and durable T-cell responses to HIV-1 antigens, but the double deletion mutant showed enhanced magnitude and quality of HIV-1 adaptive and memory responses. Our observations are relevant in the immune evaluation of MVA-B and on improvements of MVA vectors as HIV-1 vaccines.

Show MeSH
Related in: MedlinePlus