Molecular and Cellular Biochemistry

, Volume 359, Issue 1–2, pp 359–368 | Cite as

Identification of TLR inducing Th1-responsive Leishmania donovani amastigote-specific antigens

  • Ankita Srivastava
  • Nisha Singh
  • Manish Mishra
  • Vinod Kumar
  • Jalaj K. Gour
  • Surabhi Bajpai
  • Sangram Singh
  • Haushila P. Pandey
  • Rakesh K. Singh
Article

Abstract

Leishmania is known to elicit Th2 response that causes leishmaniasis progression; on the other hand, Th1 cytokines restricts amastigote growth and disease progression. In this study, we report the potential of two leishmanial antigens (65 and 98 kDa, in combination) which enhance strong macrophage effector functions, viz., production of respiratory burst enzymes, nitric oxide, and Th1 cytokines. The identification of antigens were done by resolving the crude soluble antigens on SDS-PAGE and eluted by reverse staining method. Further, RAW264.7 macrophages were challenged with eluted antigens, and the innate immune response was observed by detecting respiratory burst enzymes, nitric oxide (NOx), TNF-α, IFN-γ, IL-12, toll-like receptors (TLRs) gene expression, and TLR-signaling proteins. These antigens increased the production of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, superoxide dismutase, NOx, TNF-α, IFN-γ, IL-12, TLR2, and p38 mitogen-activated protein kinase. These antigens also induced human peripheral blood mononuclear cells proliferation and Th1 cytokine production. This study concludes that these antigens induce innate immune response as well as have prophylactic efficacy.

Keywords

Leishmaniasis Antigens Toll-like receptors Th1 cytokines 

Notes

Acknowledgments

Financial assistance received from the Department of Science and Technology (SR/FT/LS/066/2007), and the Department of Biotechnology—DBT (BT/PR11177/MED/29/99/2008), New Delhi, India is gratefully acknowledged. AS and NS are highly thankful to the DBT, New Delhi for senior research fellowship.

Conflict of interest

None.

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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Ankita Srivastava
    • 1
  • Nisha Singh
    • 1
  • Manish Mishra
    • 1
  • Vinod Kumar
    • 1
  • Jalaj K. Gour
    • 1
  • Surabhi Bajpai
    • 1
  • Sangram Singh
    • 2
  • Haushila P. Pandey
    • 1
  • Rakesh K. Singh
    • 1
  1. 1.Department of Biochemistry, Faculty of ScienceBanaras Hindu UniversityVaranasiIndia
  2. 2.Department of BiochemistryDr. R.M.L. Avadh UniversityFaizabadIndia

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