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Development of a Multivalent Subunit Vaccine against Tularemia Using Tobacco Mosaic Virus (TMV) Based Delivery System.

Banik S, Mansour AA, Suresh RV, Wykoff-Clary S, Malik M, McCormick AA, Bakshi CS - PLoS ONE (2015)

Bottom Line: Results from this study demonstrate that TMV can be used as a carrier for effective delivery of multiple F. tularensis antigens.This study provides a proof-of-concept that TMV can serve as a suitable platform for simultaneous delivery of multiple protective antigens of F. tularensis.Refinement of vaccine formulations coupled with TMV-targeting strategies developed in this study will provide a platform for development of an effective tularemia subunit vaccine as well as a vaccination approach that may broadly be applicable to many other bacterial pathogens.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, New York Medical College, Valhalla, New York, United States of America.

ABSTRACT
Francisella tularensis is a facultative intracellular pathogen, and is the causative agent of a fatal human disease known as tularemia. F. tularensis is classified as a Category A Biothreat agent by the CDC based on its use in bioweapon programs by several countries in the past and its potential to be used as an agent of bioterrorism. No licensed vaccine is currently available for prevention of tularemia. In this study, we used a novel approach for development of a multivalent subunit vaccine against tularemia by using an efficient tobacco mosaic virus (TMV) based delivery platform. The multivalent subunit vaccine was formulated to contain a combination of F. tularensis protective antigens: OmpA-like protein (OmpA), chaperone protein DnaK and lipoprotein Tul4 from the highly virulent F. tularensis SchuS4 strain. Two different vaccine formulations and immunization schedules were used. The immunized mice were challenged with lethal (10xLD100) doses of F. tularensis LVS on day 28 of the primary immunization and observed daily for morbidity and mortality. Results from this study demonstrate that TMV can be used as a carrier for effective delivery of multiple F. tularensis antigens. TMV-conjugate vaccine formulations are safe and multiple doses can be administered without causing any adverse reactions in immunized mice. Immunization with TMV-conjugated F. tularensis proteins induced a strong humoral immune response and protected mice against respiratory challenges with very high doses of F. tularensis LVS. This study provides a proof-of-concept that TMV can serve as a suitable platform for simultaneous delivery of multiple protective antigens of F. tularensis. Refinement of vaccine formulations coupled with TMV-targeting strategies developed in this study will provide a platform for development of an effective tularemia subunit vaccine as well as a vaccination approach that may broadly be applicable to many other bacterial pathogens.

No MeSH data available.


Related in: MedlinePlus

OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Monoconjugate and TMV-Multiconjugate Vaccines using Schedule I of Immunization.F. tularensis SchuS4 recombinant proteins OmpA, DnaK and Tul4 specific IgG antibody levels on day 28 in serum samples of C57BL/6 mice immunized with TMV-monoconjugate and TMV-multiconjugate vaccine using Schedule I were determined by ELISA. Serum samples obtained from naïve mice or those inoculated with TMV alone were used as controls. The data are represented as Mean ±S.D. of absorbance values measured at 450nm. Table shows comparison of antibody titers between groups of mice vaccinated with these vaccine formulations.
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pone.0130858.g007: OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Monoconjugate and TMV-Multiconjugate Vaccines using Schedule I of Immunization.F. tularensis SchuS4 recombinant proteins OmpA, DnaK and Tul4 specific IgG antibody levels on day 28 in serum samples of C57BL/6 mice immunized with TMV-monoconjugate and TMV-multiconjugate vaccine using Schedule I were determined by ELISA. Serum samples obtained from naïve mice or those inoculated with TMV alone were used as controls. The data are represented as Mean ±S.D. of absorbance values measured at 450nm. Table shows comparison of antibody titers between groups of mice vaccinated with these vaccine formulations.

Mentions: We first investigated the antibody response in mice that received TMV-monoconjugate vaccine formulation in which all the three recombinant proteins were conjugated to a single TMV virion, and received boosters only on day 7 and 14 (Schedule I). Mice were bled on day 28 post-immunization and antibody responses were determined. Higher levels of Francisella specific total IgG levels were detected in TMV-monoconjugate vaccinated mice (Fig 6). Determination of IgG isotypes on day 28 post-immunization revealed that mice vaccinated with TMV-monoconjugate vaccine induced higher IgG1 levels. However, very low to undetectable levels of IgG2a and IgG2b antibodies were observed in this group of vaccinated mice (Fig 6). Antigen specific ELISA indicated that antibodies were generated against OmpA, DnaK and Tul4 proteins (Fig 7). Collectively, these results indicated that a weak antibody response predominated by a Th2 biased immune response is generated in mice immunized using Schedule I vaccination regimen with TMV-monoconjugate vaccine formulation.


Development of a Multivalent Subunit Vaccine against Tularemia Using Tobacco Mosaic Virus (TMV) Based Delivery System.

Banik S, Mansour AA, Suresh RV, Wykoff-Clary S, Malik M, McCormick AA, Bakshi CS - PLoS ONE (2015)

OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Monoconjugate and TMV-Multiconjugate Vaccines using Schedule I of Immunization.F. tularensis SchuS4 recombinant proteins OmpA, DnaK and Tul4 specific IgG antibody levels on day 28 in serum samples of C57BL/6 mice immunized with TMV-monoconjugate and TMV-multiconjugate vaccine using Schedule I were determined by ELISA. Serum samples obtained from naïve mice or those inoculated with TMV alone were used as controls. The data are represented as Mean ±S.D. of absorbance values measured at 450nm. Table shows comparison of antibody titers between groups of mice vaccinated with these vaccine formulations.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476615&req=5

pone.0130858.g007: OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Monoconjugate and TMV-Multiconjugate Vaccines using Schedule I of Immunization.F. tularensis SchuS4 recombinant proteins OmpA, DnaK and Tul4 specific IgG antibody levels on day 28 in serum samples of C57BL/6 mice immunized with TMV-monoconjugate and TMV-multiconjugate vaccine using Schedule I were determined by ELISA. Serum samples obtained from naïve mice or those inoculated with TMV alone were used as controls. The data are represented as Mean ±S.D. of absorbance values measured at 450nm. Table shows comparison of antibody titers between groups of mice vaccinated with these vaccine formulations.
Mentions: We first investigated the antibody response in mice that received TMV-monoconjugate vaccine formulation in which all the three recombinant proteins were conjugated to a single TMV virion, and received boosters only on day 7 and 14 (Schedule I). Mice were bled on day 28 post-immunization and antibody responses were determined. Higher levels of Francisella specific total IgG levels were detected in TMV-monoconjugate vaccinated mice (Fig 6). Determination of IgG isotypes on day 28 post-immunization revealed that mice vaccinated with TMV-monoconjugate vaccine induced higher IgG1 levels. However, very low to undetectable levels of IgG2a and IgG2b antibodies were observed in this group of vaccinated mice (Fig 6). Antigen specific ELISA indicated that antibodies were generated against OmpA, DnaK and Tul4 proteins (Fig 7). Collectively, these results indicated that a weak antibody response predominated by a Th2 biased immune response is generated in mice immunized using Schedule I vaccination regimen with TMV-monoconjugate vaccine formulation.

Bottom Line: Results from this study demonstrate that TMV can be used as a carrier for effective delivery of multiple F. tularensis antigens.This study provides a proof-of-concept that TMV can serve as a suitable platform for simultaneous delivery of multiple protective antigens of F. tularensis.Refinement of vaccine formulations coupled with TMV-targeting strategies developed in this study will provide a platform for development of an effective tularemia subunit vaccine as well as a vaccination approach that may broadly be applicable to many other bacterial pathogens.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, New York Medical College, Valhalla, New York, United States of America.

ABSTRACT
Francisella tularensis is a facultative intracellular pathogen, and is the causative agent of a fatal human disease known as tularemia. F. tularensis is classified as a Category A Biothreat agent by the CDC based on its use in bioweapon programs by several countries in the past and its potential to be used as an agent of bioterrorism. No licensed vaccine is currently available for prevention of tularemia. In this study, we used a novel approach for development of a multivalent subunit vaccine against tularemia by using an efficient tobacco mosaic virus (TMV) based delivery platform. The multivalent subunit vaccine was formulated to contain a combination of F. tularensis protective antigens: OmpA-like protein (OmpA), chaperone protein DnaK and lipoprotein Tul4 from the highly virulent F. tularensis SchuS4 strain. Two different vaccine formulations and immunization schedules were used. The immunized mice were challenged with lethal (10xLD100) doses of F. tularensis LVS on day 28 of the primary immunization and observed daily for morbidity and mortality. Results from this study demonstrate that TMV can be used as a carrier for effective delivery of multiple F. tularensis antigens. TMV-conjugate vaccine formulations are safe and multiple doses can be administered without causing any adverse reactions in immunized mice. Immunization with TMV-conjugated F. tularensis proteins induced a strong humoral immune response and protected mice against respiratory challenges with very high doses of F. tularensis LVS. This study provides a proof-of-concept that TMV can serve as a suitable platform for simultaneous delivery of multiple protective antigens of F. tularensis. Refinement of vaccine formulations coupled with TMV-targeting strategies developed in this study will provide a platform for development of an effective tularemia subunit vaccine as well as a vaccination approach that may broadly be applicable to many other bacterial pathogens.

No MeSH data available.


Related in: MedlinePlus