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HSP70 domain II of Mycobacterium tuberculosis modulates immune response and protective potential of F1 and LcrV antigens of Yersinia pestis in a mouse model.

Batra L, Verma SK, Nagar DP, Saxena N, Pathak P, Pant SC, Tuteja U - PLoS Negl Trop Dis (2014)

Bottom Line: A significant difference was noticed in the expression level of IL-2, IFN-γ and TNF-α in splenocytes of immunized animals.We investigated whether the F1, LcrV and HSP70(II) antigens alone or in combination can effectively protect immunized animals from any histopathological changes.A significant difference was observed in the expression of IL-2, IFN-γ, TNF-α, and CD4+/CD8+ T cells secreting IFN-γ in the F1+LcrV+HSP70(II) vaccinated group in comparison to the F1+LcrV vaccinated group.

View Article: PubMed Central - PubMed

Affiliation: Microbiology Division, Defence Research & Development Establishment, Gwalior, India.

ABSTRACT
No ideal vaccine exists to control plague, a deadly dangerous disease caused by Yersinia pestis. In this context, we cloned, expressed and purified recombinant F1, LcrV antigens of Y. pestis and heat shock protein70 (HSP70) domain II of M. tuberculosis in E. coli. To evaluate the protective potential of each purified protein alone or in combination, Balb/C mice were immunized. Humoral and cell mediated immune responses were evaluated. Immunized animals were challenged with 100 LD50 of Y. pestis via intra-peritoneal route. Vaccine candidates i.e., F1 and LcrV generated highly significant titres of anti-F1 and anti-LcrV IgG antibodies. A significant difference was noticed in the expression level of IL-2, IFN-γ and TNF-α in splenocytes of immunized animals. Significantly increased percentages of CD4+ and CD8+ T cells producing IFN-γ in spleen of vaccinated animals were observed in comparison to control group by flow cytometric analysis. We investigated whether the F1, LcrV and HSP70(II) antigens alone or in combination can effectively protect immunized animals from any histopathological changes. Signs of histopathological lesions noticed in lung, liver, kidney and spleen of immunized animals on 3rd day post challenge whereas no lesions in animals that survived to day 20 post-infection were observed. Immunohistochemistry showed bacteria in lung, liver, spleen and kidney on 3rd day post-infection whereas no bacteria was observed on day 20 post-infection in surviving animals in LcrV, LcrV+HSP70(II), F1+LcrV, and F1+LcrV+HSP70(II) vaccinated groups. A significant difference was observed in the expression of IL-2, IFN-γ, TNF-α, and CD4+/CD8+ T cells secreting IFN-γ in the F1+LcrV+HSP70(II) vaccinated group in comparison to the F1+LcrV vaccinated group. Three combinations that included LcrV+HSP70(II), F1+LcrV or F1+LcrV+HSP70(II) provided 100% protection, whereas LcrV alone provided only 75% protection. These findings suggest that HSP70(II) of M. tuberculosis can be a potent immunomodulator for F1 and LcrV containing vaccine candidates against plague.

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Immunohistochemistry (IHC) staining for localization of Y. pestis in the organs collected from immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged.The F1 antigen of Y. pestis was identified with anti-mouse FITC conjugated secondary antibody in the tissue sections collected from immunized animal groups on 3rd day post infection including naive control i.e., Naive control (A); PBS control (B); HSP70(II) (C); F1 (D); F1+HSP70(II) (E); LcrV (F); LcrV+HSP70(II) (G); F1+LcrV (H); F1+LcrV+HSP70(II) (I). Tissue sections were collected from the survived animal groups on 20th day post infection i.e., LcrV (J); LcrV+HSP70(II) (K); F1+LcrV (L); F1+LcrV+HSP70(II) (M). Fluorescent images representing the localization of Y. pestis in tissue sections of Lung [a]; Spleen [b]; Kidney [c]; and Liver [d].
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pntd-0003322-g008: Immunohistochemistry (IHC) staining for localization of Y. pestis in the organs collected from immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged.The F1 antigen of Y. pestis was identified with anti-mouse FITC conjugated secondary antibody in the tissue sections collected from immunized animal groups on 3rd day post infection including naive control i.e., Naive control (A); PBS control (B); HSP70(II) (C); F1 (D); F1+HSP70(II) (E); LcrV (F); LcrV+HSP70(II) (G); F1+LcrV (H); F1+LcrV+HSP70(II) (I). Tissue sections were collected from the survived animal groups on 20th day post infection i.e., LcrV (J); LcrV+HSP70(II) (K); F1+LcrV (L); F1+LcrV+HSP70(II) (M). Fluorescent images representing the localization of Y. pestis in tissue sections of Lung [a]; Spleen [b]; Kidney [c]; and Liver [d].

Mentions: To study the dissemination of Y. pestis from peritoneal cavity to various vital organs i.e., lung, spleen, liver and kidney by immunohistochemistry were performed. The immunized animals including PBS control were sacrificed on day 3 after infection to localize Y. pestis by immunohistochemistry in lung, spleen, liver and kidney (Figure 8). No bacterium was observed in lung, liver, spleen and kidney isolated from the naive control group where as the clumping of Y. pestis was observed from all the vaccinated animals including control group by immunohistochemistry (Figure 8). On day 20 after infection, survived animals from LcrV; LcrV+HSP70(II); F1+LcrV and F1+LcrV+HSP70(II) groups were sacrificed for bacterial localization. No bacterial presence was observed in any of the survivors by immunohistochemistry (Fig. 8).


HSP70 domain II of Mycobacterium tuberculosis modulates immune response and protective potential of F1 and LcrV antigens of Yersinia pestis in a mouse model.

Batra L, Verma SK, Nagar DP, Saxena N, Pathak P, Pant SC, Tuteja U - PLoS Negl Trop Dis (2014)

Immunohistochemistry (IHC) staining for localization of Y. pestis in the organs collected from immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged.The F1 antigen of Y. pestis was identified with anti-mouse FITC conjugated secondary antibody in the tissue sections collected from immunized animal groups on 3rd day post infection including naive control i.e., Naive control (A); PBS control (B); HSP70(II) (C); F1 (D); F1+HSP70(II) (E); LcrV (F); LcrV+HSP70(II) (G); F1+LcrV (H); F1+LcrV+HSP70(II) (I). Tissue sections were collected from the survived animal groups on 20th day post infection i.e., LcrV (J); LcrV+HSP70(II) (K); F1+LcrV (L); F1+LcrV+HSP70(II) (M). Fluorescent images representing the localization of Y. pestis in tissue sections of Lung [a]; Spleen [b]; Kidney [c]; and Liver [d].
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256173&req=5

pntd-0003322-g008: Immunohistochemistry (IHC) staining for localization of Y. pestis in the organs collected from immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged.The F1 antigen of Y. pestis was identified with anti-mouse FITC conjugated secondary antibody in the tissue sections collected from immunized animal groups on 3rd day post infection including naive control i.e., Naive control (A); PBS control (B); HSP70(II) (C); F1 (D); F1+HSP70(II) (E); LcrV (F); LcrV+HSP70(II) (G); F1+LcrV (H); F1+LcrV+HSP70(II) (I). Tissue sections were collected from the survived animal groups on 20th day post infection i.e., LcrV (J); LcrV+HSP70(II) (K); F1+LcrV (L); F1+LcrV+HSP70(II) (M). Fluorescent images representing the localization of Y. pestis in tissue sections of Lung [a]; Spleen [b]; Kidney [c]; and Liver [d].
Mentions: To study the dissemination of Y. pestis from peritoneal cavity to various vital organs i.e., lung, spleen, liver and kidney by immunohistochemistry were performed. The immunized animals including PBS control were sacrificed on day 3 after infection to localize Y. pestis by immunohistochemistry in lung, spleen, liver and kidney (Figure 8). No bacterium was observed in lung, liver, spleen and kidney isolated from the naive control group where as the clumping of Y. pestis was observed from all the vaccinated animals including control group by immunohistochemistry (Figure 8). On day 20 after infection, survived animals from LcrV; LcrV+HSP70(II); F1+LcrV and F1+LcrV+HSP70(II) groups were sacrificed for bacterial localization. No bacterial presence was observed in any of the survivors by immunohistochemistry (Fig. 8).

Bottom Line: A significant difference was noticed in the expression level of IL-2, IFN-γ and TNF-α in splenocytes of immunized animals.We investigated whether the F1, LcrV and HSP70(II) antigens alone or in combination can effectively protect immunized animals from any histopathological changes.A significant difference was observed in the expression of IL-2, IFN-γ, TNF-α, and CD4+/CD8+ T cells secreting IFN-γ in the F1+LcrV+HSP70(II) vaccinated group in comparison to the F1+LcrV vaccinated group.

View Article: PubMed Central - PubMed

Affiliation: Microbiology Division, Defence Research & Development Establishment, Gwalior, India.

ABSTRACT
No ideal vaccine exists to control plague, a deadly dangerous disease caused by Yersinia pestis. In this context, we cloned, expressed and purified recombinant F1, LcrV antigens of Y. pestis and heat shock protein70 (HSP70) domain II of M. tuberculosis in E. coli. To evaluate the protective potential of each purified protein alone or in combination, Balb/C mice were immunized. Humoral and cell mediated immune responses were evaluated. Immunized animals were challenged with 100 LD50 of Y. pestis via intra-peritoneal route. Vaccine candidates i.e., F1 and LcrV generated highly significant titres of anti-F1 and anti-LcrV IgG antibodies. A significant difference was noticed in the expression level of IL-2, IFN-γ and TNF-α in splenocytes of immunized animals. Significantly increased percentages of CD4+ and CD8+ T cells producing IFN-γ in spleen of vaccinated animals were observed in comparison to control group by flow cytometric analysis. We investigated whether the F1, LcrV and HSP70(II) antigens alone or in combination can effectively protect immunized animals from any histopathological changes. Signs of histopathological lesions noticed in lung, liver, kidney and spleen of immunized animals on 3rd day post challenge whereas no lesions in animals that survived to day 20 post-infection were observed. Immunohistochemistry showed bacteria in lung, liver, spleen and kidney on 3rd day post-infection whereas no bacteria was observed on day 20 post-infection in surviving animals in LcrV, LcrV+HSP70(II), F1+LcrV, and F1+LcrV+HSP70(II) vaccinated groups. A significant difference was observed in the expression of IL-2, IFN-γ, TNF-α, and CD4+/CD8+ T cells secreting IFN-γ in the F1+LcrV+HSP70(II) vaccinated group in comparison to the F1+LcrV vaccinated group. Three combinations that included LcrV+HSP70(II), F1+LcrV or F1+LcrV+HSP70(II) provided 100% protection, whereas LcrV alone provided only 75% protection. These findings suggest that HSP70(II) of M. tuberculosis can be a potent immunomodulator for F1 and LcrV containing vaccine candidates against plague.

Show MeSH
Related in: MedlinePlus