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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-Multiconjugate Vaccines using Schedule I and II 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-multiconjugate vaccine using Schedule II were determined by ELISA. The plates were coated with recombinant F. tularensis SchuS4 proteins. 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. The comparisons are shown with the data obtained from mice immunized with TMV-multiconjugate vaccine using schedule I (shown in Fig 7). Table shows comparison of antibody titers between groups of mice vaccinated with Schedule I and II vaccination regimens.
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pone.0130858.g009: OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Multiconjugate Vaccines using Schedule I and II 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-multiconjugate vaccine using Schedule II were determined by ELISA. The plates were coated with recombinant F. tularensis SchuS4 proteins. 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. The comparisons are shown with the data obtained from mice immunized with TMV-multiconjugate vaccine using schedule I (shown in Fig 7). Table shows comparison of antibody titers between groups of mice vaccinated with Schedule I and II vaccination regimens.

Mentions: We further investigated if an aggressive immunization schedule consisting of TMV-multi conjugate formulation administered by both the i.n. and s.c. routes with multiple booster vaccinations (Schedule II) improves the antibody response compared to the other two vaccination strategies. It was observed that the total IgG and IgG1 antibody responses did not differ from mice vaccinated with TMV-multiconjugate vaccine using Schedule I (Fig 8). However, the levels of antibody isotypes IgG2a and IgG2b were in fact significantly higher in mice vaccinated with the TMV-multiconjugate vaccine receiving Schedule I than those receiving Schedule II vaccinations (Fig 8). We further investigated if there are any differences in levels of antibodies generated against native proteins of F. tularensis SchuS4 and F. tularensis LVS. No differences in IgG, IgG1, IgG2a and IgG2b antibody levels were observed when ELISAs were performed using F. tularensis SchuS4 and LVS lysates (Fig 8). These results indicate that similar to the results obtained with western blot analysis, antibodies from TMV-multiconjugate vaccine are equally capable of recognizing native proteins of both F. tularensis SchuS4 and F. tularensis LVS. The group of mice receiving Schedule II of TMV-multiconjugate vaccine showed significantly higher titers of OmpA antibodies, while titers of DnaK and Tul4 antibodies were similar to those receiving Schedule I vaccination with TMV-multiconjugate vaccine (Fig 9).


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-Multiconjugate Vaccines using Schedule I and II 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-multiconjugate vaccine using Schedule II were determined by ELISA. The plates were coated with recombinant F. tularensis SchuS4 proteins. 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. The comparisons are shown with the data obtained from mice immunized with TMV-multiconjugate vaccine using schedule I (shown in Fig 7). Table shows comparison of antibody titers between groups of mice vaccinated with Schedule I and II vaccination regimens.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0130858.g009: OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Multiconjugate Vaccines using Schedule I and II 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-multiconjugate vaccine using Schedule II were determined by ELISA. The plates were coated with recombinant F. tularensis SchuS4 proteins. 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. The comparisons are shown with the data obtained from mice immunized with TMV-multiconjugate vaccine using schedule I (shown in Fig 7). Table shows comparison of antibody titers between groups of mice vaccinated with Schedule I and II vaccination regimens.
Mentions: We further investigated if an aggressive immunization schedule consisting of TMV-multi conjugate formulation administered by both the i.n. and s.c. routes with multiple booster vaccinations (Schedule II) improves the antibody response compared to the other two vaccination strategies. It was observed that the total IgG and IgG1 antibody responses did not differ from mice vaccinated with TMV-multiconjugate vaccine using Schedule I (Fig 8). However, the levels of antibody isotypes IgG2a and IgG2b were in fact significantly higher in mice vaccinated with the TMV-multiconjugate vaccine receiving Schedule I than those receiving Schedule II vaccinations (Fig 8). We further investigated if there are any differences in levels of antibodies generated against native proteins of F. tularensis SchuS4 and F. tularensis LVS. No differences in IgG, IgG1, IgG2a and IgG2b antibody levels were observed when ELISAs were performed using F. tularensis SchuS4 and LVS lysates (Fig 8). These results indicate that similar to the results obtained with western blot analysis, antibodies from TMV-multiconjugate vaccine are equally capable of recognizing native proteins of both F. tularensis SchuS4 and F. tularensis LVS. The group of mice receiving Schedule II of TMV-multiconjugate vaccine showed significantly higher titers of OmpA antibodies, while titers of DnaK and Tul4 antibodies were similar to those receiving Schedule I vaccination with TMV-multiconjugate vaccine (Fig 9).

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