Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi
Objective: To evaluate the immunostimulatory potential of cross-reactive molecule heat shock protein 60 (HSP60) of filarial parasite Brugia malayi and Leishmania donovani. Methods: HSP60 of Brugia malayi (BmHSP60) was amplified using gene-specific primer, cloned in pTriEx4 vector, expressed in BL21-...
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Wolters Kluwer Medknow Publications
2021
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ftdoajarticles:oai:doaj.org/article:09671915b3074b11abc31e536dc6d4b6 2023-05-15T15:11:08+02:00 Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi Vikas Kushwaha Sukhbir Kaur 2021-01-01T00:00:00Z https://doi.org/10.4103/2221-1691.300728 https://doaj.org/article/09671915b3074b11abc31e536dc6d4b6 EN eng Wolters Kluwer Medknow Publications http://www.apjtb.org/article.asp?issn=2221-1691;year=2021;volume=11;issue=1;spage=20;epage=28;aulast=Kushwaha https://doaj.org/toc/2221-1691 https://doaj.org/toc/2588-9222 2221-1691 2588-9222 doi:10.4103/2221-1691.300728 https://doaj.org/article/09671915b3074b11abc31e536dc6d4b6 Asian Pacific Journal of Tropical Biomedicine, Vol 11, Iss 1, Pp 20-28 (2021) brugia malayi heat shock protein 60 leishmania donovani th1 cytokines th2 cytokines Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 article 2021 ftdoajarticles https://doi.org/10.4103/2221-1691.300728 2022-12-31T00:28:41Z Objective: To evaluate the immunostimulatory potential of cross-reactive molecule heat shock protein 60 (HSP60) of filarial parasite Brugia malayi and Leishmania donovani. Methods: HSP60 of Brugia malayi (BmHSP60) was amplified using gene-specific primer, cloned in pTriEx4 vector, expressed in BL21-DE3 cells, and recombinant HSP60 (rHSP60) of ~65 kDa was purified by affinity chromatography using Ni-NTA column. The recombinant protein was desalted by the dialysis membrane, and the presence of endotoxin level was determined by Limulus amebocyte lysate assay. The recombinant protein was tested for cell proliferation, nitric oxide release, expression of Th1 and Th2 cytokines, and transcription factors (STATs) in vitro using murine macrophage cell line (J774A.1). Results: Higher cell proliferation indicated that BmHSP60 had immunostimulatory potential. rBmHSP60 exposure upregulated the expression of iNOS, STAT1, STAT4, Th1 cytokines (IFN–γ, TNF–α, IL-12), and nitric oxide release. In addition, no remarkable change was observed in the expression of IL-6, IL-10, and STAT3 in macrophage cell line J774A.1. The ELISA analysis showed the levels of IFN-γ, TNF-α, and IL-12 were upregulated while IL-10 level was downregulated, revealing that BmHSP60 triggered a Th1 immune response. Conclusions: Our study demonstrates that rBmHSP60 has immunogenic properties which effectively enhances the Th1 type immune responses, and can be used as an immunoprophylactic agent against leishmaniasis. Furthermore, in vivo studies are in progress to determine the protective role of rBmHSP60 against Leishmania donovani infection in a mouse model. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Asian Pacific Journal of Tropical Biomedicine 11 1 20 |
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ftdoajarticles |
language |
English |
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brugia malayi heat shock protein 60 leishmania donovani th1 cytokines th2 cytokines Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 |
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brugia malayi heat shock protein 60 leishmania donovani th1 cytokines th2 cytokines Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 Vikas Kushwaha Sukhbir Kaur Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi |
topic_facet |
brugia malayi heat shock protein 60 leishmania donovani th1 cytokines th2 cytokines Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 |
description |
Objective: To evaluate the immunostimulatory potential of cross-reactive molecule heat shock protein 60 (HSP60) of filarial parasite Brugia malayi and Leishmania donovani. Methods: HSP60 of Brugia malayi (BmHSP60) was amplified using gene-specific primer, cloned in pTriEx4 vector, expressed in BL21-DE3 cells, and recombinant HSP60 (rHSP60) of ~65 kDa was purified by affinity chromatography using Ni-NTA column. The recombinant protein was desalted by the dialysis membrane, and the presence of endotoxin level was determined by Limulus amebocyte lysate assay. The recombinant protein was tested for cell proliferation, nitric oxide release, expression of Th1 and Th2 cytokines, and transcription factors (STATs) in vitro using murine macrophage cell line (J774A.1). Results: Higher cell proliferation indicated that BmHSP60 had immunostimulatory potential. rBmHSP60 exposure upregulated the expression of iNOS, STAT1, STAT4, Th1 cytokines (IFN–γ, TNF–α, IL-12), and nitric oxide release. In addition, no remarkable change was observed in the expression of IL-6, IL-10, and STAT3 in macrophage cell line J774A.1. The ELISA analysis showed the levels of IFN-γ, TNF-α, and IL-12 were upregulated while IL-10 level was downregulated, revealing that BmHSP60 triggered a Th1 immune response. Conclusions: Our study demonstrates that rBmHSP60 has immunogenic properties which effectively enhances the Th1 type immune responses, and can be used as an immunoprophylactic agent against leishmaniasis. Furthermore, in vivo studies are in progress to determine the protective role of rBmHSP60 against Leishmania donovani infection in a mouse model. |
format |
Article in Journal/Newspaper |
author |
Vikas Kushwaha Sukhbir Kaur |
author_facet |
Vikas Kushwaha Sukhbir Kaur |
author_sort |
Vikas Kushwaha |
title |
Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi |
title_short |
Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi |
title_full |
Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi |
title_fullStr |
Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi |
title_full_unstemmed |
Immunostimulatory role of rBmHSP60 from filarial parasite Brugia malayi |
title_sort |
immunostimulatory role of rbmhsp60 from filarial parasite brugia malayi |
publisher |
Wolters Kluwer Medknow Publications |
publishDate |
2021 |
url |
https://doi.org/10.4103/2221-1691.300728 https://doaj.org/article/09671915b3074b11abc31e536dc6d4b6 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Asian Pacific Journal of Tropical Biomedicine, Vol 11, Iss 1, Pp 20-28 (2021) |
op_relation |
http://www.apjtb.org/article.asp?issn=2221-1691;year=2021;volume=11;issue=1;spage=20;epage=28;aulast=Kushwaha https://doaj.org/toc/2221-1691 https://doaj.org/toc/2588-9222 2221-1691 2588-9222 doi:10.4103/2221-1691.300728 https://doaj.org/article/09671915b3074b11abc31e536dc6d4b6 |
op_doi |
https://doi.org/10.4103/2221-1691.300728 |
container_title |
Asian Pacific Journal of Tropical Biomedicine |
container_volume |
11 |
container_issue |
1 |
container_start_page |
20 |
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1766342039712038912 |