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Viswas Konasagara Nagaleekar - One of the best experts on this subject based on the ideXlab platform.

  • Recombinant transferrin binding protein a rtbpa fragments of pasteurella multocida serogroup b 2 provide variable protection following homologous challenge in mouse model
    Research in Veterinary Science, 2015
    Co-Authors: Sathish Bhadravati Shivachandra, Revanaiah Yogisharadhya, Abhinendra Kumar, Nihar Nalini Mohanty, Viswas Konasagara Nagaleekar
    Abstract:

    Transferrin binding protein A (TbpA), an iron acquisition surface protein that also acts as virulence factor, is widely distributed among strains of Pasteurella multocida. In the present study, a total of seven clones of TbpA fragments (39D to F777; 39D to Q697; 188V to F777; 188V to Q697; 39D to P377; 188V to P377 and 39D to F187) belonging to P. multocida B:2 were constructed, over-expressed and purified as Recombinant Fusion Proteins from Escherichia coli using affinity chromatography. Immunization of mice with rTbpA fragments resulted in a significant (p < 0.05) rise in antigen specific serum total IgG and subtypes (IgG1 and IgG2a) tires. All immunized mice challenged with 8 LD50 of P. multocida B:2 resulted in a variable protective efficacy up to 50%. The study indicated the potential possibilities to incorporate full length TbpA in subunit vaccine formulation composed of synergistic subunit antigens against haemorrhagic septicaemia (HS) in cattle and buffalo.

  • Recombinant transferrin binding protein A (rTbpA) fragments of Pasteurella multocida serogroup B:2 provide variable protection following homologous challenge in mouse model.
    Research in Veterinary Science, 2014
    Co-Authors: Sathish Bhadravati Shivachandra, Revanaiah Yogisharadhya, Abhinendra Kumar, Nihar Nalini Mohanty, Viswas Konasagara Nagaleekar
    Abstract:

    Transferrin binding protein A (TbpA), an iron acquisition surface protein that also acts as virulence factor, is widely distributed among strains of Pasteurella multocida. In the present study, a total of seven clones of TbpA fragments (39D to F777; 39D to Q697; 188V to F777; 188V to Q697; 39D to P377; 188V to P377 and 39D to F187) belonging to P. multocida B:2 were constructed, over-expressed and purified as Recombinant Fusion Proteins from Escherichia coli using affinity chromatography. Immunization of mice with rTbpA fragments resulted in a significant (p 

Sathish Bhadravati Shivachandra - One of the best experts on this subject based on the ideXlab platform.

  • Recombinant transferrin binding protein a rtbpa fragments of pasteurella multocida serogroup b 2 provide variable protection following homologous challenge in mouse model
    Research in Veterinary Science, 2015
    Co-Authors: Sathish Bhadravati Shivachandra, Revanaiah Yogisharadhya, Abhinendra Kumar, Nihar Nalini Mohanty, Viswas Konasagara Nagaleekar
    Abstract:

    Transferrin binding protein A (TbpA), an iron acquisition surface protein that also acts as virulence factor, is widely distributed among strains of Pasteurella multocida. In the present study, a total of seven clones of TbpA fragments (39D to F777; 39D to Q697; 188V to F777; 188V to Q697; 39D to P377; 188V to P377 and 39D to F187) belonging to P. multocida B:2 were constructed, over-expressed and purified as Recombinant Fusion Proteins from Escherichia coli using affinity chromatography. Immunization of mice with rTbpA fragments resulted in a significant (p < 0.05) rise in antigen specific serum total IgG and subtypes (IgG1 and IgG2a) tires. All immunized mice challenged with 8 LD50 of P. multocida B:2 resulted in a variable protective efficacy up to 50%. The study indicated the potential possibilities to incorporate full length TbpA in subunit vaccine formulation composed of synergistic subunit antigens against haemorrhagic septicaemia (HS) in cattle and buffalo.

  • Recombinant transferrin binding protein A (rTbpA) fragments of Pasteurella multocida serogroup B:2 provide variable protection following homologous challenge in mouse model.
    Research in Veterinary Science, 2014
    Co-Authors: Sathish Bhadravati Shivachandra, Revanaiah Yogisharadhya, Abhinendra Kumar, Nihar Nalini Mohanty, Viswas Konasagara Nagaleekar
    Abstract:

    Transferrin binding protein A (TbpA), an iron acquisition surface protein that also acts as virulence factor, is widely distributed among strains of Pasteurella multocida. In the present study, a total of seven clones of TbpA fragments (39D to F777; 39D to Q697; 188V to F777; 188V to Q697; 39D to P377; 188V to P377 and 39D to F187) belonging to P. multocida B:2 were constructed, over-expressed and purified as Recombinant Fusion Proteins from Escherichia coli using affinity chromatography. Immunization of mice with rTbpA fragments resulted in a significant (p 

Lidice Bernardo - One of the best experts on this subject based on the ideXlab platform.

  • Serotype-specificity of Recombinant Fusion Proteins containing domain III of dengue virus.
    Virus research, 2008
    Co-Authors: Alienys Izquierdo, Lidice Bernardo, Jorge Martin, Emidalys Santana, Lisset Hermida, Gerardo Guillén, María G Guzmán
    Abstract:

    Here, the antigenic specificity of the Recombinant Fusion Proteins containing aa 286-426 of the dengue envelope protein fused to P64k from Neisseria meningitidis and the cross-reactive antibody response induced in immunized mice and monkeys were evaluated. The anti-dengue mice antibodies showed a higher reactivity to the homologous Recombinant Proteins compared to the wide cross-reactivity observed by dot blot to the viral antigens. The immune response induced by the Recombinant Proteins in mice and monkeys, was highly serotype specific. The serotype-specificity associated with these Recombinant Proteins in addition to the high antigenicity, immunogenicity and protecting capacity suggest their advantages as possible vaccine candidates.

  • Immunogenicity and protective efficacy of a Recombinant Fusion protein containing the domain III of the dengue 1 envelope protein in non-human primates
    Antiviral research, 2008
    Co-Authors: Lidice Bernardo, Alienys Izquierdo, Emidalys Santana, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Delfina Rosario, Jorge Jiménez Castro, Lisset Hermida
    Abstract:

    Recombinant Fusion Proteins containing the aa 286-426 of the dengue envelope protein fused to P64k protein from Neisseria meningitidis have been previously reported. Particularly, the immunogenicity and protective capacity of the dengue 2 Recombinant protein was demonstrated in Macaca fascicularis monkeys. Here we evaluate the Recombinant Fusion protein containing the domain III of the dengue 1 envelope protein (PD10) in non-human primates (M. fascicularis and rhesus monkeys) and compare the effect of aluminum hydroxide and Freund adjuvant on the immunity induced. The PD10 protein emulsified in Freund adjuvant was highly immunogenic in M. fascicularis and rhesus monkeys. Following dengue 1 virus challenge, animals immunized with PD10 in Freund adjuvant were protected from viremia. However, monkeys receiving PD10 in aluminum hydroxide developed a poor antibody response and were not protected from viral challenge. These preliminary experiments are encouraging. Other formulations or vaccine schedules are being studied in an attempt to find regimens that enhance immunological protection.

  • Anamnestic antibody response after viral challenge in monkeys immunized with dengue 2 Recombinant Fusion Proteins
    Archives of Virology, 2008
    Co-Authors: Lidice Bernardo, Jorge Martin, Lisset Hermida, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Rosmari Rodríguez-roche, Aida Zulueta, Laura Lazo
    Abstract:

    The suitability of dengue 2 envelope domain III Recombinant Fusion Proteins [(Fusion (PD5) and insertion (PD3) variants)] for inducing functional antibodies and a protective immune response in nonhuman primates has been reported. However, the evaluation of the antibody response after immunization did not correlate with the protection data as measured by viremia detection. Here, we characterized the anamnestic immune response after viral challenge in monkeys immunized with the dengue 2 Recombinant Proteins in an attempt to define correlates of protection useful for vaccine studies. Monkeys immunized with PD5 (most protected group) exhibited an earlier increase in the anti-DENV-2 IgM response after challenge compared to control animals. Hemagglutination-inhibiting (HAI) antibodies were increased significantly earlier in PD5-immunized animals compared to those immunized with PD3. The fully protected monkeys showed the earliest HAI antibody response. These results underline the usefulness of the anamnestic antibody response for supporting protection data. The induction of an early HAI and IgM antibody response after challenge suggest a protective role against dengue virus (DENV) infection in monkeys, supporting their use as correlates of protection in vaccine studies.

  • Anamnestic antibody response after viral challenge in monkeys immunized with dengue 2 Recombinant Fusion Proteins
    Archives of Virology, 2008
    Co-Authors: Lidice Bernardo, Jorge Martin, Lisset Hermida, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Rosmari Rodríguez-roche, Aida Zulueta, Laura Lazo
    Abstract:

    The suitability of dengue 2 envelope domain III Recombinant Fusion Proteins [(Fusion (PD5) and insertion (PD3) variants)] for inducing functional antibodies and a protective immune response in nonhuman primates has been reported. However, the evaluation of the antibody response after immunization did not correlate with the protection data as measured by viremia detection. Here, we characterized the anamnestic immune response after viral challenge in monkeys immunized with the dengue 2 Recombinant Proteins in an attempt to define correlates of protection useful for vaccine studies. Monkeys immunized with PD5 (most protected group) exhibited an earlier increase in the anti-DENV-2 IgM response after challenge compared to control animals. Hemagglutination-inhibiting (HAI) antibodies were increased significantly earlier in PD5-immunized animals compared to those immunized with PD3. The fully protected monkeys showed the earliest HAI antibody response. These results underline the usefulness of the anamnestic antibody response for supporting protection data. The induction of an early HAI and IgM antibody response after challenge suggest a protective role against dengue virus (DENV) infection in monkeys, supporting their use as correlates of protection in vaccine studies.

  • A Recombinant Fusion protein containing the domain III of the dengue-2 envelope protein is immunogenic and protective in nonhuman primates
    Vaccine, 2006
    Co-Authors: Lisset Hermida, Lidice Bernardo, Jorge Martin, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Beatriz Sierra, R. Rodríguez, Aida Zulueta
    Abstract:

    We have previously reported the construction and evaluation in mice of Recombinant Fusion Proteins formed by a fragment (aa 286-426) of the dengue envelope protein and the P64k protein from Neisseria meningitidis. In this work we describe the immunization of Macaca fascicularis monkeys with two variants of these Proteins [PD3 (insertion variant) and PD5 (Fusion variant)] corresponding to serotype 2. Four doses of the Proteins adjuvated in Freund's adjuvant were administered and the kinetics of antibody induction was monitored by ELISA and neutralization tests. Monkeys receiving PD3 or PD5 developed functional antibodies (Abs) in a dose-dependent manner. Following challenge with 5 log PFU of wild type dengue-2 virus (DEN2), animals immunized with PD5 were protected from developing viremia. These results constitute a proof-of-concept demonstrating that a fragment of the dengue envelope protein, containing the domain III and produced as a Recombinant Fusion protein in Escherichia coli, induces functional and protective immunity in a nonhuman primate model.

Snezana Djordjevic - One of the best experts on this subject based on the ideXlab platform.

  • Crystal Structure of Tobacco Etch Virus Protease Shows the Protein C Terminus Bound within the Active Site
    Journal of molecular biology, 2005
    Co-Authors: Christine M. Nunn, Mark Jeeves, Matthew J. Cliff, Gillian T. Urquhart, Roger George, Luke H Chao, Yugo Tscuchia, Snezana Djordjevic
    Abstract:

    Tobacco etch virus (TEV) protease is a cysteine protease exhibiting stringent sequence specificity. The enzyme is widely used in biotechnology for the removal of the affinity tags from Recombinant Fusion Proteins. Crystal structures of two TEV protease mutants as complexes with a substrate and a product peptide provided the first insight into the mechanism of substrate specificity of this enzyme. We now report a 2.7A crystal structure of a full-length inactive C151A mutant protein crystallised in the absence of peptide. The structure reveals the C terminus of the protease bound to the active site. In addition, we determined dissociation constants of TEV protease substrate and product peptides using isothermal titration calorimetry for various forms of this enzyme. Data suggest that TEV protease could be inhibited by the peptide product of autolysis. Separate modes of recognition for native substrates and the site of TEV protease self-cleavage are proposed.

Lisset Hermida - One of the best experts on this subject based on the ideXlab platform.

  • Serotype-specificity of Recombinant Fusion Proteins containing domain III of dengue virus.
    Virus research, 2008
    Co-Authors: Alienys Izquierdo, Lidice Bernardo, Jorge Martin, Emidalys Santana, Lisset Hermida, Gerardo Guillén, María G Guzmán
    Abstract:

    Here, the antigenic specificity of the Recombinant Fusion Proteins containing aa 286-426 of the dengue envelope protein fused to P64k from Neisseria meningitidis and the cross-reactive antibody response induced in immunized mice and monkeys were evaluated. The anti-dengue mice antibodies showed a higher reactivity to the homologous Recombinant Proteins compared to the wide cross-reactivity observed by dot blot to the viral antigens. The immune response induced by the Recombinant Proteins in mice and monkeys, was highly serotype specific. The serotype-specificity associated with these Recombinant Proteins in addition to the high antigenicity, immunogenicity and protecting capacity suggest their advantages as possible vaccine candidates.

  • Immunogenicity and protective efficacy of a Recombinant Fusion protein containing the domain III of the dengue 1 envelope protein in non-human primates
    Antiviral research, 2008
    Co-Authors: Lidice Bernardo, Alienys Izquierdo, Emidalys Santana, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Delfina Rosario, Jorge Jiménez Castro, Lisset Hermida
    Abstract:

    Recombinant Fusion Proteins containing the aa 286-426 of the dengue envelope protein fused to P64k protein from Neisseria meningitidis have been previously reported. Particularly, the immunogenicity and protective capacity of the dengue 2 Recombinant protein was demonstrated in Macaca fascicularis monkeys. Here we evaluate the Recombinant Fusion protein containing the domain III of the dengue 1 envelope protein (PD10) in non-human primates (M. fascicularis and rhesus monkeys) and compare the effect of aluminum hydroxide and Freund adjuvant on the immunity induced. The PD10 protein emulsified in Freund adjuvant was highly immunogenic in M. fascicularis and rhesus monkeys. Following dengue 1 virus challenge, animals immunized with PD10 in Freund adjuvant were protected from viremia. However, monkeys receiving PD10 in aluminum hydroxide developed a poor antibody response and were not protected from viral challenge. These preliminary experiments are encouraging. Other formulations or vaccine schedules are being studied in an attempt to find regimens that enhance immunological protection.

  • Anamnestic antibody response after viral challenge in monkeys immunized with dengue 2 Recombinant Fusion Proteins
    Archives of Virology, 2008
    Co-Authors: Lidice Bernardo, Jorge Martin, Lisset Hermida, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Rosmari Rodríguez-roche, Aida Zulueta, Laura Lazo
    Abstract:

    The suitability of dengue 2 envelope domain III Recombinant Fusion Proteins [(Fusion (PD5) and insertion (PD3) variants)] for inducing functional antibodies and a protective immune response in nonhuman primates has been reported. However, the evaluation of the antibody response after immunization did not correlate with the protection data as measured by viremia detection. Here, we characterized the anamnestic immune response after viral challenge in monkeys immunized with the dengue 2 Recombinant Proteins in an attempt to define correlates of protection useful for vaccine studies. Monkeys immunized with PD5 (most protected group) exhibited an earlier increase in the anti-DENV-2 IgM response after challenge compared to control animals. Hemagglutination-inhibiting (HAI) antibodies were increased significantly earlier in PD5-immunized animals compared to those immunized with PD3. The fully protected monkeys showed the earliest HAI antibody response. These results underline the usefulness of the anamnestic antibody response for supporting protection data. The induction of an early HAI and IgM antibody response after challenge suggest a protective role against dengue virus (DENV) infection in monkeys, supporting their use as correlates of protection in vaccine studies.

  • Anamnestic antibody response after viral challenge in monkeys immunized with dengue 2 Recombinant Fusion Proteins
    Archives of Virology, 2008
    Co-Authors: Lidice Bernardo, Jorge Martin, Lisset Hermida, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Rosmari Rodríguez-roche, Aida Zulueta, Laura Lazo
    Abstract:

    The suitability of dengue 2 envelope domain III Recombinant Fusion Proteins [(Fusion (PD5) and insertion (PD3) variants)] for inducing functional antibodies and a protective immune response in nonhuman primates has been reported. However, the evaluation of the antibody response after immunization did not correlate with the protection data as measured by viremia detection. Here, we characterized the anamnestic immune response after viral challenge in monkeys immunized with the dengue 2 Recombinant Proteins in an attempt to define correlates of protection useful for vaccine studies. Monkeys immunized with PD5 (most protected group) exhibited an earlier increase in the anti-DENV-2 IgM response after challenge compared to control animals. Hemagglutination-inhibiting (HAI) antibodies were increased significantly earlier in PD5-immunized animals compared to those immunized with PD3. The fully protected monkeys showed the earliest HAI antibody response. These results underline the usefulness of the anamnestic antibody response for supporting protection data. The induction of an early HAI and IgM antibody response after challenge suggest a protective role against dengue virus (DENV) infection in monkeys, supporting their use as correlates of protection in vaccine studies.

  • A Recombinant Fusion protein containing the domain III of the dengue-2 envelope protein is immunogenic and protective in nonhuman primates
    Vaccine, 2006
    Co-Authors: Lisset Hermida, Lidice Bernardo, Jorge Martin, Mayling Alvarez, Irina Prado, Carlos López, Rafael Martínez, Beatriz Sierra, R. Rodríguez, Aida Zulueta
    Abstract:

    We have previously reported the construction and evaluation in mice of Recombinant Fusion Proteins formed by a fragment (aa 286-426) of the dengue envelope protein and the P64k protein from Neisseria meningitidis. In this work we describe the immunization of Macaca fascicularis monkeys with two variants of these Proteins [PD3 (insertion variant) and PD5 (Fusion variant)] corresponding to serotype 2. Four doses of the Proteins adjuvated in Freund's adjuvant were administered and the kinetics of antibody induction was monitored by ELISA and neutralization tests. Monkeys receiving PD3 or PD5 developed functional antibodies (Abs) in a dose-dependent manner. Following challenge with 5 log PFU of wild type dengue-2 virus (DEN2), animals immunized with PD5 were protected from developing viremia. These results constitute a proof-of-concept demonstrating that a fragment of the dengue envelope protein, containing the domain III and produced as a Recombinant Fusion protein in Escherichia coli, induces functional and protective immunity in a nonhuman primate model.