Acta Parasitologica

, Volume 58, Issue 4, pp 468–477 | Cite as

Immune responses of B. malayi thioredoxin (TRX) and venom allergen homologue (VAH) chimeric multiple antigen for lymphatic filariasis

  • Gandhirajan Anugraha
  • Parasurama Jawaharlal Jeyaprita
  • Jayaprakasam Madhumathi
  • Tamilvanan Sheeba
  • Perumal Kaliraj
Original Paper


Although multiple vaccine strategy for lymphatic filariasis has provided tremendous hope, the choice of antigens used in combination has determined its success in the previous studies. Multiple antigens comprising key vaccine candidates from different life cycle stages would provide a promising strategy if the antigenic combination is chosen by careful screening. In order to analyze one such combination, we have used a chimeric construct carrying the well studied B. malayi antigens thioredoxin (BmTRX) and venom allergen homologue (BmVAH) as a fusion protein (TV) and evaluated its immune responses in mice model. The efficacy of fusion protein vaccine was explored in comparison with the single antigen vaccines and their cocktail. In mice, TV induced significantly high antibody titer of 1,28,000 compared to cocktail vaccine TRX+VAH (50,000) and single antigen vaccine TRX (16,000) or VAH (50,000). Furthermore, TV elicited higher level of cellular proliferative response together with elevated levels of IFN-γ, IL-4 and IL-5 indicating a Th1/Th2 balanced response. The isotype antibody profile showed significantly high level of IgG1 and IgG2b confirming the balanced response elicited by TV. Immunization with TV antigen induced high levels of both humoral and cellular immune responses compared to either cocktail or antigen given alone. The result suggests that TV is highly immunogenic in mice and hence the combination needs to be evaluated for its prophylactic potential.


Lymphatic filariasis Brugia malayi thioredoxin venom allergen homologue chimeric construct multiple antigen vaccine 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anand S.B., Gnanasekar M., Thangadurai M., Prabhu P.R., Kaliraj P., Ramaswamy K. 2007. Immune response studies with Wuchereria bancrofti vespid allergen homologue (WbVAH) in human lymphatic filariasis. Parasitology Research, 101, 981–988. DOI: 10.1007/s00436-007-0571-2.PubMedCrossRefPubMedCentralGoogle Scholar
  2. Anand S.B., Kodumudi K.N., Reddy M.V., Kaliraj P. 2011. A combination of two Brugia malayi filarial vaccine candidate antigens (BmALT-2 and BmVAH) enhances immune responses and protection in jirds. Journal of Helminthology, 85, 442–452. DOI: 10.1017/S0022149X10000799.PubMedCrossRefGoogle Scholar
  3. Anand S.B., Murugan V., Prabhu P.R., Anandharaman V., Reddy M.V., Kaliraj P. 2008. Comparison of immunogenicity, protective efficacy of single and cocktail DNA vaccine of Brugia malayi abundant larval transcript (ALT-2) and thioredoxin peroxidase (TPX) in mice. Acta Tropica, 107, 106–112. DOI: 10.1016/j.actatropi ca.2008.04.018.PubMedCrossRefGoogle Scholar
  4. Babu B.V., Mishra S. 2008. Mass drug administration under the programme to eliminate lymphatic filariasis in Orissa, India: a mixed-methods study to identify factors associated with compliance and non-compliance. Transactions of the Royal Society of Tropical Medicine and Hygiene, 102, 1207–1213. DOI: 10.1016/j.trstmh.2008.05.023.PubMedCrossRefGoogle Scholar
  5. Babu S., Ganley L.M., Klei T.R., Shultz L.D., Rajan T.V. 2000. Role of gamma interferon and interleukin-4 in host defense against the human filarial parasite Brugia malayi. Infection and Immunity, 68, 3034–3035.PubMedCrossRefPubMedCentralGoogle Scholar
  6. Babu S., Shultz L.D., Klei T.R., Rajan T.V. 1999. Immunity in experimental murine filariasis: roles of T and B cells revisited. Infection and Immunity, 67, 3166–3167.PubMedPubMedCentralGoogle Scholar
  7. Carvalho-Queiroz C., Cook R., Wang C.C., Correa-Oliveira R., Bailey N.A., Egilmez N.K. 2004. Cross-reactivity of Schistosoma mansoni cytosolic superoxide dismutase, a protective vaccine candidate, with host superoxide dismutase and identification of parasite-specific B epitopes. Infection and Immunity, 72, 2635–2647.PubMedCrossRefPubMedCentralGoogle Scholar
  8. Chenthamarakshan V., Reddy M.V., Harinath B.C. 1995. Immuno prophylactic potential of a 120 kDa Brugia malayi adult antigen fraction, BmA-2, in lymphatic filariasis. Parasite Immunology, 17, 277–285.PubMedCrossRefGoogle Scholar
  9. Chiumiento L., Bruschi F. 2009. Enzymatic antioxidant systems in helminth parasites. Parasitology Research, 105, 593–603. DOI: 10.1007/s00436-009-1483-0.PubMedCrossRefGoogle Scholar
  10. Dakshinamoorthy G., Samykutty A.K., Munirathinam G., Reddy M.V., Kalyanasundaram R. 2012. Multivalent fusion protein vaccine for lymphatic filariasis. Vaccine, ? DOI: 10.1016/j.vaccine.2012.09.055.Google Scholar
  11. Dissanayake S., Perler F.B., Xu M., Southworth M.W., Yee C.K., Wang S., Dreyer G., Watawana L., Kurniawan L., Fuhrman J.A. 1995. Differential recognition of microfilarial chitinase, a transmission-blocking vaccine candidate antigen, by sera from patients with Brugian and Bancroftian filariasis. American Journal of Tropical Medicine and Hygiene, 53, 289–294.PubMedGoogle Scholar
  12. El-Setouhy M., Abd Elaziz K.M., Helmy H., Farid H.A., Kamal H.A., Ramzy R.M., Shannon W.D., Weil G.J. 2007. The effect of compliance on the impact of mass drug administration for elimination of lymphatic filariasis in Egypt. American Journal of Tropical Medicine and Hygiene, 77, 1069–1073.PubMedPubMedCentralGoogle Scholar
  13. Gnanasekar M., Rao K.V., He Y.X., Mishra P.K., Nutman T.B., Kaliraj P., Ramaswamy K. 2004. Novel phage display-based sub tractive screening to identify vaccine candidates of Brugia malayi. Infection and Immunity, 72, 4707–4715. DOI:.4707-4715.2004.PubMedCrossRefPubMedCentralGoogle Scholar
  14. Gomez-Escobar N., Bennett C., Prieto-Lafuente L., Aebischer T., Blackburn C.C., Maizels R.M. 2005. Heterologous expression of the filarial nematode alt gene products reveals their potential to inhibit immune function. BMC Biology, 23, 3–8. DOI: 10.1186/1741-7007-3-8.Google Scholar
  15. Gozar M.M., Price V.L., Kaslow D.C. 1998. Saccharomyces cerevisiae-secreted fusion proteins Pfs25 and Pfs28 elicit potent Plasmodium falciparum transmission-blocking antibodies in mice. Infection and Immunity, 66, 59–64.PubMedPubMedCentralGoogle Scholar
  16. Hager D.A., Burgess R.R. 1980. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Analytical Biochemistry, 109, 76–86.PubMedCrossRefGoogle Scholar
  17. Hawdon J.M., Jones B.F., Hoffman D.R., Hotez P.J. 1996. Cloning and characterization of Ancylostoma secreted protein. A novel protein associated with the transition to parasitism by infective hookworm larvae. Journal of Biological Chemistry, 271, 6672–6678.PubMedCrossRefGoogle Scholar
  18. Henkle-Duhrsen K., Kampkotter A. 2001. Antioxidant enzyme families in parasitic nematodes. Molecular and Biochemical Parasitology, 114, 129–142.PubMedCrossRefGoogle Scholar
  19. Hoerauf A., Satoguina J., Saeftel M., Specht S. 2005. Immuno-modulation by filarial nematodes. Parasite Immunology, 27, 417–429.PubMedCrossRefGoogle Scholar
  20. Horton J. 2009. The development of albendazole for lymphatic filariasis. Annals of Tropical Medicine and Parasitology, 103, 33–40. DOI: 10.1179/0003 49809 X12502035776595.CrossRefGoogle Scholar
  21. Hotez P.J. 2009. Mass drug administration and integrated control for the world’s high-prevalence neglected tropical diseases, 85, 659–664. DOI: 10.1038/clpt.2009.16.Google Scholar
  22. Kalyanasundaram R., Balumuri P. 2011. Multivalent vaccine formulation with BmVAL-1 and BmALT-2 confer significant protection against challenge infections with Brugia malayi in mice and jirds. Research and Reports in Tropical Medicine, 2, 45–56. DOI: 10.2147/RRTM.S13679.CrossRefGoogle Scholar
  23. Kunchithapautham K., Padmavathi B., Narayanan R.B., Kaliraj P., Scott A.L. 2003. Thioredoxin from Brugia malayi: defining a 16-kilodalton class of thioredoxins from nematodes. Infection and Immunity, 71, 4119–4126.PubMedCrossRefPubMedCentralGoogle Scholar
  24. Laemmli U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.PubMedCrossRefGoogle Scholar
  25. Lange A.M., Yutanawiboonchai W., Scott P., Abraham, D. 1994. IL-4-and IL-5-dependent protective immunity to Onchocerca volvulus infective larvae in BALB/cBYJ mice. Journal of Immunology, 153, 205–211.Google Scholar
  26. Lawrence R.A. 2001. Immunity to filarial nematodes. Veterinary Parasitology, 100(1–2), 33–44.PubMedCrossRefGoogle Scholar
  27. LoVerde P.T., Carvalho-Queiroz C., Cook R. 2004. Vaccination with antioxidant enzymes confers protective immunity against challenge infection with Schistosoma mansoni. Memórias do Instituto Oswaldo Cruz, 99, 37–43. DOI: S0074-02762004000900007.PubMedCrossRefGoogle Scholar
  28. Madhumathi J., Prince P.R., Anugraha G., Kiran P., Rao D.N., Reddy M.V. Kaliraj P. 2010. Identification and characterization of nematode specific protective epitopes of Brugia malayi TRX towards development of synthetic vaccine construct for lymphatic filariasis. Vaccine, 28, 5038–5048.PubMedCrossRefGoogle Scholar
  29. Maizels R.M., Blaxter M.L., Scott A.L. 2001. Immunological genomics of Brugia malayi: filarial genes implicated in immune evasion and protective immunity. Parasite Immunology, 23, 327–344.PubMedCrossRefGoogle Scholar
  30. Mazumdar S., Mukherjee P., Yazdani S.S., Jain S.K., Mohmmed A., Chauhan V.S. 2010. Plasmodium falciparum merozoite surface protein 1 (MSP-1)-MSP-3 chimeric protein: immunogenicity determined with human-compatible adjuvants and induction of protective immune response. Infection and Immunity, 78, 872–883. DOI: 10.1128/IAI.00427-09.PubMedCrossRefPubMedCentralGoogle Scholar
  31. Murray J., Gregory W.F., Gomez-Escobar N., Atmadja A.K., Maizels R.M. 2001. Expression and immune recognition of Brugia malayi VAL-1, a homologue of vespid venom allergens and Ancylostoma secreted proteins. Molecular Biochemical Parasitology, 118, 89–96.PubMedCrossRefGoogle Scholar
  32. Peters B., Sidney J., Bourne P., Bui H.H., Buus S., Doh G. 2005. The design and implementation of the immune epitope data base and analysis resource. Immunogenetics, 57, 326–336. DOI: 10.1007/s00251-005-0803-5.PubMedCrossRefGoogle Scholar
  33. Samykutty A., Dakshinamoorthy G., Kalyanasundaram R. 2010. Multivalent Vaccine for Lymphatic Filariasis. Procedia in Vaccinology, 3, 12–18. DOI: 10.1016/j.provac.2010.11.003.PubMedCrossRefPubMedCentralGoogle Scholar
  34. Schwab A.E., Churcher T.S., Schwab A.J., Basanez M.G., Prichard R.K. 2007. An analysis of the population genetics of potential multi-drug resistance in Wuchereria bancrofti due to combination chemotherapy. Parasitology, 134, 1025–1040. DOI: 10.1017/S0031182007002363.PubMedCrossRefGoogle Scholar
  35. Tawe W., Pearlman E., Unnasch T.R., Lustigman S. 2000. Angiogenic activity of Onchocerca volvulus recombinant proteins similar to vespid venom antigen 5. Molecular Biochemical Parasitology, 109, 91–99.PubMedCrossRefGoogle Scholar
  36. Thirugnanam S., Pandiaraja P., Ramaswamy K., Murugan V., Gnanasekar M., Nandakumar K., Reddy M.V., Kaliraj P. 2007. Brugia malayi: comparison of protective immune responses induced by Bm-alt-2 DNA, recombinant Bm-ALT-2 protein and prime-boost vaccine regimens in a jird model. Experimental Parasitology, 116, 483–491. DOI: 10.1016/j.expp ar a.2007.02./017.PubMedCrossRefPubMedCentralGoogle Scholar
  37. Towbin H., Staehelin T., Gordon J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, 76, 4350–4354.CrossRefGoogle Scholar
  38. Vanam U., Pandey V., Prabhu P.R., Dakshinamurthy G., Reddy M.V., Kaliraj P. 2009. Evaluation of immunoprophylactic efficacy of Brugia malayi transglutaminase (BmTGA) in single and multiple antigen vaccination with BmALT-2 and BmTPX for human lymphatic filariasis. American Journal of Tropical Medicine and Hygiene, 80, 319–324.PubMedGoogle Scholar
  39. Vincent A.L., Sodeman W.A., Winters A. 1980. Development of Brugia pahangi in normal and nude mice. Journal of Parasitology, 66, 448.PubMedCrossRefGoogle Scholar

Copyright information

© Versita Warsaw and Springer-Verlag Wien 2013

Authors and Affiliations

  • Gandhirajan Anugraha
    • 1
  • Parasurama Jawaharlal Jeyaprita
    • 1
  • Jayaprakasam Madhumathi
    • 1
  • Tamilvanan Sheeba
    • 1
  • Perumal Kaliraj
    • 1
  1. 1.Centre for BiotechnologyAnna UniversityChennaiIndia

Personalised recommendations