Iranian Journal of Parasitology 2016. 11(4):480-488.

Modulation of the Immune Response to DNA Vaccine Encoding Gene of 8-kDa Subunit of Echinococcus granulosus Antigen B Using Murine Interleukin-12 Plasmid in BALB/c Mice


Background: The current study was designed to evaluate immune responses induced by DNA vaccines encoding 8-kDa subunit of antigen B (HydI) of Echinococcus granulosus and murine interleukin 12 (IL-12) as genetic adjuvants in BALB/c mice.

Methods: Expression plasmid pcDNA3.1 containing HydI (pcHyd1) as vaccine along with the murine interleukin 12 (pcMIL12) as adjuvant were used. Thirty-five mice in the five experimental groups received PBS, empty pcDNA3.1, pcHydІ, pcMIL-12, and pcHydІ+ pcMIL-12 in days zero, 14th and 28th. Two weeks after the last immunization, evaluation of the immune response was performed by evaluating the proliferation of splenic lymphocytes, IFN-γ and IL-4, determination of IgG isotyping titer.

Results: Mice that received the pcHydI+pcMIL12 exhibited higher levels of lymphocyte proliferation compared to mice that received the pcHydI alone (P<0.001), and produced significantly more IFN-γ in comparison to other groups (P< 0.001). In addition, they produced significantly less IL-4 than mice receiving the PBS and the empty plasmid (P<0.023). The IgG2a levels were clearly higher in pcHydI+pcMIL12 group in comparison with the groups of pcHydI alone, empty plasmid, and PBS. In contrast, IgG1 was elevated in the group of pcHydI.

Conclusion: Co-delivery of IL-12 with DNA encoding 8-kDa subunit of antigen B was effective significantly in inducing the immune response in mice.


Hydatid disease; Th1/Th2; Cytokines; Immunization;Genetic adjuvant; Antigen B

Full Text:



Pakala T, Molina M,Wu GY. Hepatic Echinococcal Cysts: A Review. J Clin Transl Hepatol. 2016; 4(1):39-46.

Qingling M, Guanglei W, Jun Q, Xinquan Z, Tianli L, Xuemei S, et al. Prevalence of hydatid cysts in livestock animals in Xinjiang, China. Korean J Parasitol. 2014; 52(3):331-4.

Dakkak A. Echinococcosis/hydatidosis: a severe threat in Mediterranean countries. Vet Parasitol. 2010; 174(1):2-11.

Mandal S,Mandal MD. Human cystic echinococcosis: epidemiologic, zoonotic, clinical, diagnostic and therapeutic aspects. Asian Pac J Trop Med. 2012; 5(4):253-60.

Torgerson P. Economic effects of echinococcosis. Acta Tropica. 2003; 85(2):113-18.

Torgerson P, Budke C. Echinococcosis–an international public health challenge. Res Vet Sci. 2003; 74(3):191-202.

Heath D, Yang W, Li T, Xiao Y, Chen X, Huang Y, et al. Control of hydatidosis. Parasitol Int. 2006; 55:47-52.

Lightowlers MW,Gauci CG. Vaccines against cysticercosis and hydatidosis. Vet Parasitol 2001; 101(3):337-52.

Lightowlers MW. Vaccines against cysticercosis and hydatidosis: foundations in taeniid cestode immunology. Parasitol Inte. 2006; 55:39-43.

Craig PS, McManus DP, Lightowlers MW, Chabalgoity JA, Garcia HH, Gavidia CM, et al. Prevention and control of cystic echinococcosis. Lancet Infect Dis. 2007; 7(6):385-94.

Torgerson PR. Mathematical models for the control of cystic echinococcosis. Parasito Inte. 2006; 55(2):53-58.

Torgerson P. The use of mathematical models to simulate control options for echinococcosis. Acta Tropica. 2003; 85(2):211-21.

Khan KH. DNA vaccines: roles against diseases. Germs. 2013; 3(1):26-35.

Chow C, Gauci CG, Cowman AF, Lightowlers MW. A gene family expressing a host-protective antigen of Echinococcus granulosus. Mol Biochem Parasitol. 2001; 118(1):83-88.

Mamuti W, Sako Y, Bart J-M, Nakao M, Ma X, Wen H, et al. Molecular characterization of a novel gene encoding an 8-kDa-subunit of antigen B from Echinococcus granulosus genotypes 1 and 6. Parasitol Inte. 2007; 56(4):313-16.

Azizi H, Kazemi B, Bandehpour M, Mohebali M, Khamesipour A, Aryaeipour M, et al. Molecular Cloning and Expression an 8-kDa Subunit of Antigen B from G1 strain of Echinococcus granulosus. Iran J Public Health. 2015; 44(7):962-8.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth. 1983; 65(1-2):55-63.

Bansal A, Paliwal PK, Sagi SS,Sairam M. Effect of adjuvants on immune response and protective immunity elicited by recombinant Hsp60 (GroEL) of Salmonella typhi against S. typhi infection. Mol Cell Biochem. 2010; 337(1-2):213-21.

Woollard DJ, Gauci CG, Heath DD,Lightowlers MW. Protection against hydatid disease induced with the EG95 vaccine is associated with conformational epitopes. Vaccine. 2000; 19(4):498-507.

LI ZJ, WANG YN, Qi W, Wei Z. Echinococcus granulosus 14-3-3 protein: a potential vaccine candidate against challenge with Echinococcus granulosus in mice. Biomed Environ Sci. 2012; 25(3):352-58.

Chow C, Gauci CG, Vural G, Jenkins DJ, Heath DD, Rosenzvit MC, et al. Echinococcus granulosus: variability of the host-protective EG95 vaccine antigen in G6 and G7 genotypic variants. Exp Parasitol. 2008; 119(4):499-505.

Pirestani M, Dalimi A, Sarvi S, Khoramabadi N. Evaluation of Immunogenicity of Novel Isoform of EG95 (EG95-5G1) From Echinococcus granulosus in BALB/C Mice. Iran J Parasitol. 2014; 9(4):491-502.

Wedrychowicz H. Antiparasitic DNA vaccines in 21st century. Acta Parasitol. 2015; 60(2):179-89.

Ulmer JB, Donnelly JJ, Parker SE, Rhodes GH, Felgner PL, Dwarki V, et al. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science. 1993; 259(5102):1745-49.

Zhang W, Wen H, Li J, Lin R,McManus DP. Immunology and immunodiagnosis of cystic echinococcosis: an update. Clin Dev Immunol. 2011; 20(12):1-9.

Smooker PM, Kennedy NJ, Steeper KR, Christopoulos H,Spithill TW. Fasciola: kinetics and quality of humoral responses to fatty acid binding protein and cathepsin l following delivery as DNA vaccines in mice. Exp Parasitol. 2001; 97(3):154-60.

Lança ASC, de Sousa KP, Atouguia J, Prazeres DMF, Monteiro GA,Silva MS. Trypanosoma brucei: immunisation with plasmid DNA encoding invariant surface glycoprotein gene is able to induce partial protection in experimental African trypanosomiasis. Exp Parasitol. 2011; 127(1):18-24.

Yan H-K, Yuan Z-G, Petersen E, Zhang X-X, Zhou D-H, Liu Q, et al. Toxoplasma gondii: protective immunity against experimental toxoplasmosis induced by a DNA vaccine encoding the perforin-like protein 1. Exp Parasitol. 2011; 128(1):38-43.

Xue M, He S, Cui Y, Yao Y,Wang H. Evaluation of the immune response elicited by multi-antigenic DNA vaccine expressing SAG1, ROP2 and GRA2 against Toxoplasma gondii. Parasitol Int. 2008; 57(4):424-29.

Wozniak TM, Ryan AA, Triccas JA,Britton WJ. Plasmid interleukin-23 (IL-23), but not plasmid IL-27, enhances the protective efficacy of a DNA vaccine against Mycobacterium tuberculosis infection. Infect Immun. 2006; 74(1):557-65.

Calarota SA,Weiner DB. Enhancement of human immunodeficiency virus type 1‐DNA vaccine potency through incorporation of T helper 1 molecular adjuvants. Immunol Rev. 2004; 199(1):84-99.

Liu Q, Shang L, Jin H, Wei F, Zhu X-Q,Gao H. The protective effect of a Toxoplasma gondii SAG1 plasmid DNA vaccine in mice is enhanced with IL-18. Res Vet Sci. 2010; 89(1):93-97.

Wolf SF, Sieburth D,Sypek J. Interleukin 12: a key modulator of immune function. Stem Cells. 1994; 12(2):154-68.

Letscher-Bru V, Villard O, Risse B, Zauke M, Klein J-P,Kien TT. Protective effect of vaccination with a combination of recombinant surface antigen 1 and interleukin-12 against toxoplasmosis in mice. Infect Immun. 1998; 66(9):4503-06.

Xue M, He S, Zhang J, Cui Y, Yao Y,Wang H. Comparison of cholera toxin A2/B and murine interleukin-12 as adjuvants of Toxoplasma multi-antigenic SAG1-ROP2 DNA vaccine. Exp Parasitol. 2008; 119(3):352-57.

Cui Yl, He Sy, Xue Mf, Zhang J, Wang Hx,Yao Y. Protective effect of a multiantigenic DNA vaccine against Toxoplasma gondii with co‐delivery of IL‐12 in mice. Parasite Immunol. 2008; 30(5):309-13.

Chen G, Chen H, Guo H, Zheng H. Protective effect of DNA-mediated immunization with a combination of SAG1 and IL-2 gene adjuvant against infection of Toxoplasma gondii in mice. Chin Med J (Engl). 2002; 115(10):1448-52.

Schadeck EB, Sidhu M, Egan MA, Chong S-Y, Piacente P, Masood A, et al. A dose sparing effect by plasmid encoded IL-12 adjuvant on a SIVgag-plasmid DNA vaccine in rhesus macaques. Vaccine. 2006; 24(21):4677-87.

Chow Y-H, Chiang B-L, Lee Y-L, Chi W-K, Lin W-C, Chen Y-T, et al. Development of Th1 and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccines can be modulated by codelivery of various cytokine genes. J Immunol. 1998; 160(3):1320-29.


  • There are currently no refbacks.

Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.