Comparative Evaluation of Routes of Administration and Efficacy of Monovalent and Bivalent Filarial Vaccine Candidates; BmALT-2 and WbGST

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Mohini Nakhale
Ravi Shankar Yadav
Udaikumar Padigel
Kalyan Goswami
Satish Kumar


Aims: To evaluate immunoprophylactic adequacy of monovalent and bivalent filarial vaccine candidates; BmALT-2 and WbGST through different routes of immunization viz., intramuscular (i.m.), intraperitoneal (i.p.) or subcutaneous (s.c.) way.

Study Design:  In vivo and in vitro experimental studies using rodent model of filariasis.

Place and Duration of Study: Department of Biochemistry and J B Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Maharashtra, India, between June 2016 and July 2017.

Methodology: Male Mastomys (n=5-7/group) of 6-8 weeks age were immunized with 25 µg/dose of rBmALT-2, rWbGST, or rBmALT-2+rWbGST per animal along with alum as adjuvant through either of i.m., i.p. or s.c. routes. The control groups of mice received alum only. The protective immunity elicited in the animals was checked by antibody-dependent cellular cytotoxicity and micropore cytotoxicity assay. The splenocytes proliferation and cytokine profile were performed for the appraisal of cellular immune response.

Results: Our results demonstrated that immunization through i.m. route is more effective as compared to i.p. or s.c. routes and comparatively monovalent rBmALT-2 induces better protection as compared to monovalent rWbGST or bivalent dose consisting of rBmALT-2+rWbGST. Mastomys immunized through i.m. route with rBmALT-2 alone could induce 77.50% in vitro and 73.28% in vivo cytotoxicity (P<0.05) against Brugia malayi infective larvae (L3), whereas, rWbGST alone and rBmALT-2+rWbGST combined could induce 65% and 68% in vitro and 61% and 65% in vivo cytotoxicity (P<0.05), respectively. Our serological analysis showed that Mastomys immunized i.m. with rBmALT-2 or rWbGST or rBmALT-2+rWbGST induced IgG1, IgG2a and IgG3 antibody response, significantly higher than control groups. The protective effect was found to be associated with a predominantly increased level of IFN-γ and IL-4 in response to rBmALT-2.

Conclusion: Intramuscular route is the most effective way of administration of vaccine for better prophylactic response in filariasis. Additionally, co-administration of monovalent vaccine seems to interfere with the prophylactic effect of monovalent rBmALT-2 and rWbGST vaccines.

Filariasis, antibody dependent cellular cytotoxicity (ADCC), vaccination, BmALT-2, WbGST.

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Nakhale, M., Yadav, R. S., Padigel, U., Goswami, K., & Kumar, S. (2020). Comparative Evaluation of Routes of Administration and Efficacy of Monovalent and Bivalent Filarial Vaccine Candidates; BmALT-2 and WbGST. South Asian Journal of Parasitology, 4(3), 27-37. Retrieved from
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World Health Organization. Global programme to eliminate lymphatic filariasis: Progress report, 2017 WHO. WER. 2018;44(91):589-604.

Ottesen EA. Editorial: The global programme to eliminate lymphatic filariasis. Trop Med Int Health. 2000;5:591-594.

Kushwaha S, Singh PK, Rana AK, Misra-Bhattacharya S. Immunization of Mastomys coucha with Brugia malayi recombinant trehalose-6-phosphate phosphatase results in significant protection against homologous challenge infection. PloS One. 2013;8(8):72585.

Anand SB, Murugan V, Prabhu PR, Anandharaman V, Reddy MV, Kaliraj P. 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 trop. 2008;107(2):106-112.

Thirugnanam S, Pandiaraja P, Ramaswamy K, Murugan V, Gnanasekar M, Nandakumar K, et al. 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. Exp Parasitol. 2007;116(4):483-491.

Vanam U, Pandey V, Prabhu PR, Dakshinamurthy G, Reddy MV, Kaliraj P. Evaluation of immunoprophylactic efficacy of Brugia malayi transglutaminase (BmTGA) in single and multiple antigen vaccination with BmALT-2 and BmTPX for human lymphatic filariasis. Am J Trop Med Hyg. 2009;80(2):319-324.

Anand SB, Kodumudi KN, Reddy MV, Kaliraj P. A combination of two Brugia malayi filarial vaccine candidate antigens (BmALT-2 and BmVAH) enhances immune responses and protection in jirds. J Helminthol. 2011;85(4):442-452.

Dakshinamoorthy G, Samykutty AK, Munirathinam G, Reddy MV, Kalyanasundaram R. Multivalent fusion protein vaccine for lymphatic filariasis. Vaccine. 2013;31(12):1616-1622.

Gregory WF, Blaxter ML, Maizels RM. Differentially expressed, abundant trans-spliced cDNAs from larval Brugia malayi. Mol Biochem Parasitol. 1997; 87(1):85-95.

Ramachandran S, Kumar MP, Rami RM, Chinnaiah HB, Nutman T, Kaliraj P, et al. The larval specific lymphatic filarial ALT-2: Induction of protection using protein or DNA vaccination. Microbiol Immunol. 2004; 48(12):945-955.

Gupta S, Bhandari YP, Reddy MV, Harinath BC, Rathaur S. Setaria cervi: Immunoprophylactic potential of glutathione-S-transferase against filarial parasite Brugia malayi. Exp Parasitol. 2005;109(4):252-255.

Veerapathran A, Dakshinamoorthy G, Gnanasekar M, Reddy MV, Kalyanasundaram R. Evaluation of Wuchereria bancrofti GST as a vaccine candidate for lymphatic filariasis. PLoS Negl Trop Dis. 2009;3(6):457.

Gregory WF, Atmadja AK, Allen JE, Maizels RM. The abundant larval transcript-1 and-2 genes of Brugia malayi encode stage-specific candidate vaccine antigens for filariasis. Infect Immun. 2000; 68(7):4174-4179.

Gnanasekar M, Rao KV, He YX, Mishra PK, Nutman TB, Kaliraj P, et al. Novel phage display-based subtractive screening to identify vaccine candidates of Brugia malayi. Infect Immun. 2004;72(8):4707-4715.

Sexton JL, Milner AR, Panaccio MI, Waddington JO, Wijffels G, Chandler D, et al. Glutathione S-transferase. Novel vaccine against Fasciola hepatica infection in sheep. J Immunol. 1990;145(11):3905-3910.

Grzych JM, Grezel D, Xu CB, Neyrinck JL, Capron M, Ouma JH, et al. IgA antibodies to a protective antigen in human Schistosomiasis mansoni. J Immunol 1993;150(2):527-535.

Boulanger D, Trottein F, Mauny F, Bremond P, Couret D, Pierce RJ, et al. Vaccination of goats against the trematode Schistosoma bovis with a recombinant homologous schistosome-derived glutathione S-transferase. Parasite Immunol. 1994;16(8):399-406.

Morrison CA, Colin T, Sexton JL, Bowen F, Wicker J, Friedel T, et al. Protection of cattle against Fasciola hepatica infection by vaccination with glutathione S-transferase. Vaccine. 1996;14(17-18):1603-1612.

Rathaur S, Yadav M, Gupta S, Anandharaman V, Reddy MV. Filarial glutathione-S-transferase: A potential vaccine candidate against lymphatic filariasis. Vaccine. 2008;26(32):4094- 4100.

Suzuki T, Seregeg IG. A mass dissection technique for determining infectivity rate of filariasis vectors. Jpn J Exp Med. 1979; 49(2):117-121.

Office of Laboratory Animal Welfare. Public Health Service Policy on Humane Care And Use Of Laboratory Animals. Bethesda, MD: National Institutes of Health, U.S. Department of Health and Human Services; 2015. NIH publication 15-8013; 2015.

Accessed 3 August 2015 Available:

National Institutes of Health, Office of Animal Care and Use. Guidelines for preparing USDA annual reports and assigning USDA pain and distress categories; 2009.

Accessed 26 May 2010 Available:

Anugraha G, Madhumathi J, Prince PR, Jeya Prita PJ, Khatri VK, Amdare NP, et al. Chimeric epitope vaccine from multistage antigens for lymphatic filariasis. Scand J Immunol. 2015;82(4):380-389.

Chandrashekar R, Rao UR, Parab PB, Subrahmanyam D. Brugia malayi: Serum dependent cell-mediated reactions to microfilariae. Se Asian J Trop Med. 1985; 16(1):15-21.

Chandrashekar R, Rao UR, Subrahmanyam D. Serum dependent cell-mediated immune reactions to Brugia pahangi infective larvae. Parasite Immunol. 1985;7(6):633-641.

Weiss N, Tanner M. Studies on Dipetalonema viteae (Filarioidea) Antibody-dependent cell-mediated destruction of microfilariae in vivo. Z Tropenmed Parasitol. 1979;30(1):73-80.

Grezel D, Capron M, Grzych JM, Fontaine J, Lecocq JP, Capron A. Protective immunity induced in rat schistosomiasis by a single dose of the Sm28GST recombinant antigen: Effector mechanisms involving IgE and IgA antibodies. Eur J Immunol. 1993;23(2):454-460.

Mitchell GF. Glutathione S-transferases-potential components of anti-schistosome vaccines? Parasito Today. 1989;5(2):34-37.

Andure D, Pote K, Khatri V, Amdare N, Padalkar R, Reddy MV. Immunization with Wuchereria bancrofti glutathione-S-transferase elicits a mixed Th1/Th2 type of protective immune response against filarial infection in Mastomys. Indian J Clin Biochem. 2016;31(4):423-430.

Mehta K, Sindhu RK, Subrahmanyam D, Nelson DS. IgE-dependent adherence and cytotoxicity of rat spleen and peritoneal cells to Litomosoide scarinii microfilariae. Clin Exp Immunol. 1980;41(1):107.

Samykutty A, Dakshinamoorthy G, Kalyanasundaram R. Multivalent vaccine for lymphatic filariasis. Procedia Vaccinol. 2010;3:12-18.

Kalyanasundaram R, Balumuri P. Multivalent vaccine formulation with BmVAL-1 and BmALT-2 confer significant protection against challenge infections with Brugia malayi in mice and jirds. Res Rep Trop Med. 2011;2:45.

Abraham D, Grieve RB, Mika-Grieve M, Seibert BP. Active and passive immunization of mice against larval Dirofilaria immitis. J Parasitol. 1988;74: 275-282.

Shi Q, Lynch MM, Romero M, Burns JM. Enhanced protection against malaria by a chimeric merozoite surface protein vaccine. Infect. Immun. 2007;75(3):1349-58.

Dabir S, Dabir P, Goswamy K, Reddy MV. Prophylactic evaluation of recombinant extracellular superoxide dismutase of Brugia malayi in jird model. Vaccine. 2008; 26(29-30):3705-3710.

Rizzo LV, DeKruyff RH, Umetsu DT. Generation of B cell memory and affinity maturation. Induction with Th1 and Th2 T cell clones. J Immunol. 1992;148(12): 3733-3739.

Akiyama Y, Lubeck MD, Steplewski Z, Koprowski H. Induction of mouse IgG2a-and IgG3-dependent cellular cytotoxicity in human monocytic cells (U937) by immune interferon. Cancer Res. 1984;44(11):5127-5131.

Lawrence RA. Immunity to filarial nematodes. Vet Parasitol. 2001;100(1-2):33-44.

Leu RW, Robinson CJ, Wiggins JA, Shannon BJ, Rummage JA, Horn MJ. Photometric assays for FcRI-dependent binding, phagocytosis, and antibody-dependent cellular cytotoxicity mediated by monomeric IgGγ2a in murine peritoneal macrophages. J Immunol Methods. 1988; 113(2):269-278.

Murray HW, Andre L M, Lu CM, DeVecchio JL, Matsuhashi M, Ma X, Heinzel FP. Determinants of response to interleukin-10 receptor blockade immunotherapy in experimental visceral leishmaniasis. The Journal of Infectious Diseases. 2003; 188(3):458-64.

Bogdan C, Vodovotz Y & Nathan C. Macrophage deactivation by interleukin 10. J Exp Med 1991;174:1549–1555.

Gazzinelli RT, Oswald IP, James SL & Sher A. IL-10 inhibits parasite killing and nitrogen oxide production by IFN-gamma-activated macrophages. J Immunol 1992; 148:1792–1796.

Oswald IP, Wynn TA, Sher A, James SL. Interleukin 10 inhibits macrophage microbicidal activity by blocking the endogenous production of tumor necrosis factor alpha required as a costimulatory factor for interferon gamma-induced activation. Proc Natl Acad Sci U S A. 1992;89:8676–8680.

Bhattacharyya S, Ghosh S, Jhonson PL, Bhattacharya SK & Majumdar S. Immunomodulatory role of interleukin-10 in visceral leishmaniasis: Defective activation of protein kinase C-mediated signal transduction events. Infect Immun. 2001; 69:1499–1507.

Kane MM & Mosser DM. The role of IL-10 in promoting disease progression in leishmaniasis. J Immunol. 2001;166:1141–1147.

Silva JS, Morrissey PJ, Grabstein KH, Mohler KM, Anderson D & Reed SG. Interleukin 10 and interferon gamma regulation of experimental Trypanosoma cruzi infection. J Exp Med. 1992;175:169–174.

Wilson MS, Cheever AW, White SD, Thompson RW & Wynn TA. IL-10 blocks the development of resistance to re-infection with Schistosoma mansoni. PLoS Pathog. 2011;7:1002171.

McKee AS & Pearce EJ. CD25+ CD4+ cells contribute to Th2 polarization during helminths infection by suppressing Th1 response development. J Immunol. 2004; 173:1224–1231

Brunet LR, Finkelman FD, Cheever AW, Kopf MA, Pearce EJ. IL-4 protects against TNF-alpha-mediated cachexia and death during acute schistosomiasis. J Immuno. 1997;159(2):777-785.

Martin C, Al-Qaoud KM, Ungeheuer MN, Paehle K, Vuong PN, Bain O, et al. IL-5 is essential for vaccine-induced protection and for resolution of primary infection in murine filariasis. Med Microbiol Immunol. 2000;189(2):67-74.

Else KJ, Finkelman FD. Invited review intestinal nematode parasites, cytokines and effector mechanisms. Int J Parasitol. 1998;28(8):1145-1158.