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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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Histopathology of the organs collected from the immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged with Y. pestis.Tissue sections were stained with hematoxylin and eosin for pathological examination. Tissue section collected from naive control and immunized animals on 3rd day post infection 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). Photomicrograph represents the histopathology of Lung[a]: the arrows in the panel B indicate the infiltration of neutrophils. Photomicrograph of spleen [b]: in the panel B, reduced density of white pulp follicle and congestion in the red pulp, lymphoid follicle depletion shown by arrow and the presence of megakaryocytes shown by bold arrow. Photomicrograph of kidney [c]: the granular degeneration of parenchyma was observed in the panel B (bold arrows) and swelling in renal tubules (arrows). Photomicrograph of liver [d]: in the panel B, the hepatocytes degeneration was observed as indicated by arrow.
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pntd-0003322-g007: Histopathology of the organs collected from the immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged with Y. pestis.Tissue sections were stained with hematoxylin and eosin for pathological examination. Tissue section collected from naive control and immunized animals on 3rd day post infection 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). Photomicrograph represents the histopathology of Lung[a]: the arrows in the panel B indicate the infiltration of neutrophils. Photomicrograph of spleen [b]: in the panel B, reduced density of white pulp follicle and congestion in the red pulp, lymphoid follicle depletion shown by arrow and the presence of megakaryocytes shown by bold arrow. Photomicrograph of kidney [c]: the granular degeneration of parenchyma was observed in the panel B (bold arrows) and swelling in renal tubules (arrows). Photomicrograph of liver [d]: in the panel B, the hepatocytes degeneration was observed as indicated by arrow.

Mentions: The animals sacrificed on day 3 after infection, histopathological lesions were observed in the lung tissues (Figure 7a). Normal alveolar pattern with normal alveolar septa, air duct, alveoli and bronchioles with intact epithelium were observed from naive control group (Figure 7a [A]) whereas all the vaccinated including control group, lung parenchyma showed inflammation including neutrophil infiltration into the airways and alveoli as shown by arrow (Figure 7a [B]). The significant lung lesions were congestion, hemorrhage, granulovacuolar degeneration of bronchiole associated lymphoid tissue, bronchial lumen occlusion and psuedomembrane formation (Figure 7a [B-I]). Survived animals from LcrV; LcrV+HSP70(II); F1+LcrV and F1+LcrV+HSP70(II) vaccinated groups effectively recovered as no histopathological lesions were observed (Figure 7a [J-M]).


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)

Histopathology of the organs collected from the immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged with Y. pestis.Tissue sections were stained with hematoxylin and eosin for pathological examination. Tissue section collected from naive control and immunized animals on 3rd day post infection 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). Photomicrograph represents the histopathology of Lung[a]: the arrows in the panel B indicate the infiltration of neutrophils. Photomicrograph of spleen [b]: in the panel B, reduced density of white pulp follicle and congestion in the red pulp, lymphoid follicle depletion shown by arrow and the presence of megakaryocytes shown by bold arrow. Photomicrograph of kidney [c]: the granular degeneration of parenchyma was observed in the panel B (bold arrows) and swelling in renal tubules (arrows). Photomicrograph of liver [d]: in the panel B, the hepatocytes degeneration was observed as indicated by arrow.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003322-g007: Histopathology of the organs collected from the immunized group animals on 3rd and 20th day post infection with Y. pestis and the naive control animals that were neither immunized nor challenged with Y. pestis.Tissue sections were stained with hematoxylin and eosin for pathological examination. Tissue section collected from naive control and immunized animals on 3rd day post infection 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). Photomicrograph represents the histopathology of Lung[a]: the arrows in the panel B indicate the infiltration of neutrophils. Photomicrograph of spleen [b]: in the panel B, reduced density of white pulp follicle and congestion in the red pulp, lymphoid follicle depletion shown by arrow and the presence of megakaryocytes shown by bold arrow. Photomicrograph of kidney [c]: the granular degeneration of parenchyma was observed in the panel B (bold arrows) and swelling in renal tubules (arrows). Photomicrograph of liver [d]: in the panel B, the hepatocytes degeneration was observed as indicated by arrow.
Mentions: The animals sacrificed on day 3 after infection, histopathological lesions were observed in the lung tissues (Figure 7a). Normal alveolar pattern with normal alveolar septa, air duct, alveoli and bronchioles with intact epithelium were observed from naive control group (Figure 7a [A]) whereas all the vaccinated including control group, lung parenchyma showed inflammation including neutrophil infiltration into the airways and alveoli as shown by arrow (Figure 7a [B]). The significant lung lesions were congestion, hemorrhage, granulovacuolar degeneration of bronchiole associated lymphoid tissue, bronchial lumen occlusion and psuedomembrane formation (Figure 7a [B-I]). Survived animals from LcrV; LcrV+HSP70(II); F1+LcrV and F1+LcrV+HSP70(II) vaccinated groups effectively recovered as no histopathological lesions were observed (Figure 7a [J-M]).

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