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Parasitology Research

, Volume 118, Issue 6, pp 1849–1863 | Cite as

Troponin 1 of human filarial parasite Brugia malayi: cDNA cloning, expression, purification, and its immunoprophylactic potential

  • Vikas Kushwaha
  • Prachi Tewari
  • Payal Mandal
  • Anurag Tripathi
  • P. Kalpana MurthyEmail author
Immunology and Host-Parasite Interactions - Original Paper

Abstract

In the search for immunoprophylactics for the control of human lymphatic filariasis, we recently identified troponin 1 (Tn1) in Brugia malayi adult worms. The present study reports the cloning and expression of the B. malayi Tn1 (Tn1bm), its immunoprophylactic efficacy against B. malayi infection, and the immunological responses of the host. The Tn1bm gene was cloned (Acc no. JF912447) and expressed, and the purified recombinant Tn1bm (rTn1bm) presented a single ~ 27 kDa band. Parasite load in rTn1bm-immunized BALB/c mice challenged with B. malayi infective larvae (L3) was assessed. In rTn1bm-immunized animals, IgE, IgG, and IgG subclasses in the serum, cell proliferative response, Th1 and Th2 cytokine secretion (from splenocytes), and NO release (from peritoneal macrophages) were determined. Antibody-dependent cell-mediated cytotoxicity (ADCC) to L3 was assayed using rTn1bm-immune serum. The innate immune response markers MHC class-I, MHC class-II, TLR2, TLR4, and TLR6 expression in peritoneal macrophages and CD3+, CD4+, CD8+, and CD19+ in the splenocyte population were determined in Tn1bm-exposed cells from naïve mice. rTn1bm-immunized L3-challenged animals showed a 60% reduction in parasite burden. Immunization upregulated cellular proliferation, cytokine (IFN-γ, TNF-α, IL-1β, IL-4, IL-6, and IL-10) secretion, NO release, and antigen-specific IgG, IgG1, and IgG2b antibody levels. rTn1bm-immune serum killed > 65% of L3 in the ADCC assay. Increased MHC class-II, TLR2, and TLR6 expression and the relative CD4+ and CD19+ cell populations of naïve animal cells indicated the ability of rTn1bm to mobilize innate immune responses. This is the first report of the immunoprophylactic potential of rTn1bm against B. malayi.

Keywords

Troponin 1 Brugia malayi Immunoprophylaxis Th1 and Th2 responses ADCC MHC class-I MHC class-II TLRs CD3+ CD4+ CD8+ CD19+ 

Notes

Acknowledgments

Thanks are due to the Council of Scientific and Industrial Research, New Delhi, India, for the Emeritus Scientist Project award (21(0963)/13/EMR-II; 29-10-2014) to PKM and for the Research Associateship award to PT and VK. The authors thank the Honorable Vice Chancellor, of Lucknow University, and Head of the Department of Zoology, Lucknow University, Lucknow, for providing laboratory facilities and the Director of CSIR-CDRI, Lucknow, for allowing the use of the experimental animal wing for animal experiments. The authors also thank the Director of the CSIR-Indian Institute of Toxicology Research, Lucknow, India, for allowing for part of the work to be carried out in his institute and Dr. Mukesh Srivastava, Biometry section, Division of Clinical and Experimental Medicine, CSIR-CDRI, Lucknow, for help with the statistical analyses.

Compliance with ethical standards

Ethical approval

The animal study protocol was approved by the National Laboratory Animal Centre (NLAC), CSIR-Central Drug Research Institute (CDRI), Lucknow, India, following the rules and guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Environments and Forests, Govt. of India, New Delhi, for use and handling of animals. The animals used in the study were housed in climatically controlled pathogen-free animal quarters (Temp.: 23 ± 2 °C; RH: 60%; and photoperiod: 12 h light–dark cycles) at NALC, CSIR-CDRI, Lucknow, India, and they were fed standard rodent maintenance diet (prepared in-house) and water ad libitum. The animal study protocol was approved (approval number: IAEC/2011/145 renewed dated 03/07/2014).

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Vikas Kushwaha
    • 1
    • 2
  • Prachi Tewari
    • 1
  • Payal Mandal
    • 3
  • Anurag Tripathi
    • 3
  • P. Kalpana Murthy
    • 1
    Email author
  1. 1.Department of ZoologyUniversity of LucknowLucknowIndia
  2. 2.Postdoctoral Fellow, Zoology DepartmentPanjab UniversityChandigarhIndia
  3. 3.Food Toxicology Lab, Food, Drug and Chemical Toxicology GroupCSIR-Indian Institute of Toxicology ResearchLucknowIndia

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