Volume 25, Issue 4 (7-2021)                   IBJ 2021, 25(4): 284-296 | Back to browse issues page

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Ebrahimzadeh F, Shirdast H, Taromchi A, Talebkhan Y, Haniloo A, Esmaeilzadeh A, et al . Induction of Immunogenic Response in BALB/c Mice by Live and Killed Form of Recombinant Lactococcus lactis Displaying EG95 of Echinococcus granulosus. IBJ 2021; 25 (4) :284-296
URL: http://ibj.pasteur.ac.ir/article-1-3340-en.html
Background: Cystic echinococcosis is a zoonotic parasitic infection caused by Echinococcus granulosus worldwide and is associated with economic losses among livestock animals. EG95 is an immunogenic antigen from the E. granulosus. Lactococcus lactis has been prested as a safe vehicle for antigen delivery. The goal of this study was to design a novel L. lactis strain displaying EG95 as a vaccine delivery system. Methods: The eg95 encoding gene fragment fused to the M6 anchoring protein was cloned into the pNZ7021 vector, and L. lactis NZ9000 displaying recombinant EG95 was constructed. The expression of an approximately 32-kDa EG95 protein was confirmed by Western blotting and immunofluorescence analysis. The immune responses were evaluated in BALB/c mice immunized orally and subcutaneously with the live and killed recombinant L. lactis, respectively. Results: Total IgG level in mice immunized with heat-killed recombinant L. lactis (pNZ7021-eg95) significantly increased compared to the control group. Mucosal IgA was significantly higher in mice received live recombinant L. lactis (pNZ7021-eg95) compared to the control mice. Splenic lymphocytes from immunized mice represented the high levels of IFN-γ and the low-levels of IL-4 and IL-10. Conclusion: Our results indicate that immunization with EG95-expressing L. lactis can induce both specific humoral and cellular immune responses in mice.

1. Craig PS, McManus DP, Lightowlers MW, Chabalgoity JA, Garcia HH, Gavidia CM, Gilman RH, Gonzalez AE, Lorca M, Naquira C, Nieto A, Schantz PM. Prevention and control of cystic echinococcosis. Lancet infectious disease 2007; 7(6): 385-394. [DOI:10.1016/S1473-3099(07)70134-2]
2. McManus DP, Zhang W, Li J, Bartley PB. Echinococcosis. The lancet 2003; 362(9392): 1295-1304. [DOI:10.1016/S0140-6736(03)14573-4]
3. Eckert J, Deplazes P. Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clinical microbiology reviews 2004; 17(1): 107-135. [DOI:10.1128/CMR.17.1.107-135.2004]
4. Cvejic D, Schneider C, Fourie J, de Vos C, Bonneau S, Bernachon N, Hellmann K. Efficacy of a single dose of milbemycin oxime/praziquantel combination tablets, Milpro®, against adult Echinococcus multilocularis in dogs and both adult and immature E. multilocularis in young cats. Parasitology research 2016; 115(3): 1195-1202. [DOI:10.1007/s00436-015-4855-7]
5. Chow C, Gauci CG, Cowman AF, Lightowlers MW. A gene family expressing a host-protective antigen of Echinococcus granulosus. Molecular and biochemical parasitology 2001; 118(1): 83-88. [DOI:10.1016/S0166-6851(01)00373-5]
6. Chow C, Gauci CG, Cowman AF, Lightowlers MW. Echinococcus granulosus: oncosphere-specific transcription of genes encoding a host-protective antigen. Experimental parasitology 2004; 106(3-4): 183-186. [DOI:10.1016/j.exppara.2004.03.009]
7. Lightowlers M, Lawrence S, Gauci C, Young J, Ralston M, Maas D, Heath D. Vaccination against hydatidosis using a defined recombinant antigen. Parasite immunology 1996; 18(9): 457-462. [DOI:10.1111/j.1365-3024.1996.tb01029.x]
8. Lightowlers M, Jensen O, Fernandez E, Iriarte J, Woollard D, Gauci C, Jenkins D, Heath D. Vaccination trials in Australia and Argentina confirm the effectiveness of the EG95 hydatid vaccine in sheep. International journal for parasitology 1999; 29(4): 531-534. [DOI:10.1016/S0020-7519(99)00003-X]
9. 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. Biomedical and environmental sciences 2012; 25(3): 352-358.
10. Shi Z, Wang Y, Li Z, Li Z, Bo Y, Ma R, Zhao W. Cloning, expression, and protective immunity in mice of a gene encoding the diagnostic antigen P-29 of Echinococcus granulosus. Acta biochimica et biophysica sinica 2009; 41(1): 79-85. [DOI:10.1093/abbs/gmn009]
11. Sun J, Wang Y, Li Z, Ma R, Ji H, Xiong Y, Wang Y, Li Z, Zhao W. Echinococcus granulosus: immunoprotection accompanyied by humoral and cytokine response against secondary hydatidosis in mice immunized with rEg. Myophilin. Veterinary research communications 2011; 35(4): 193-200. [DOI:10.1007/s11259-010-9456-4]
12. Behnsen J, Deriu E, Sassone-Corsi M, Raffatellu M. Probiotics: properties, examples, and specific applications. Cold spring harbor perspectives medicine 2013; 3(3): a010074. [DOI:10.1101/cshperspect.a010074]
13. Mercenier A, Muller-Alouf H, Grangette C. Lactic acid bacteria as live vaccines. Current issues in molecular biology 2000; 2: 17-26.
14. Nouaille S, Ribeiro LA, Miyoshi A, Pontes D, Le Loir Y, Oliveira SC, Langella P, Azevedo V. Heterologous protein production and delivery systems for Lactococcus lactis. Genetics and molecular research 2003; 2(1): 102-111.
15. Ma Y, Liu J, Hou J, Dong Y, Lu Y, Jin L, Cao R, Li T, Wu J. Oral administration of recombinant Lactococcus lactis expressing HSP65 and tandemly repeated P277 reduces the incidence of type I diabetes in non-obese diabetic mice. PloS one 2014; 9(8): e105701. [DOI:10.1371/journal.pone.0105701]
16. Bermúdez-Humarán LG, Nouaille S, Zilberfarb V, Corthier G, Gruss A, Langella P, Issad T. Effects of intranasal administration of a leptin-secreting Lactococcus lactis recombinant on food intake, body weight, and immune response of mice. Appllied and environmental microbiology 2007; 73(16): 5300-5307. [DOI:10.1128/AEM.00295-07]
17. Watterlot L, Rochat T, Sokol H, Cherbuy C, Bouloufa I, Lefèvre F, Gratadoux J-J, Honvo-Hueto E, Chilmonczyk S, Blugeon S, Corthier G, Lengella P, Bermudez-Humaran L. Intragastric administration of a superoxide dismutase-producing recombinant Lactobacillus casei BL23 strain attenuates DSS colitis in mice. International journal of food microbiology 2010; 144(1): 35-41. [DOI:10.1016/j.ijfoodmicro.2010.03.037]
18. Bermúdez-Humarán LG, Cortes-Perez NG, Lefèvre F, Guimarães V, Rabot S, Alcocer-Gonzalez JM, Gratadoux J-J, Rodriguez-Padilla C, Tamez-Guerra RS, Corthier G, Gruss A, Langella P. A novel mucosal vaccine based on live Lactococci expressing E7 antigen and IL-12 induces systemic and mucosal immune responses and protects mice against human papillomavirus type 16-induced tumors. Journal of immunology 2005; 175(11): 7297-7302. [DOI:10.4049/jimmunol.175.11.7297]
19. Medina E, Guzmán CA. Use of live bacterial vaccine vectors for antigen delivery: potential and limitations. Vaccine 2001; 19(13-14): 1573-1580. [DOI:10.1016/S0264-410X(00)00354-6]
20. Cano-Garrido O, Seras-Franzoso J, Garcia-Fruitos E. Lactic acid bacteria: reviewing the potential of a promising delivery live vector for biomedical purposes. Microbial cell factories 2015; 14(1): 137. [DOI:10.1186/s12934-015-0313-6]
21. Bohlul E, Hasanlou F, Taromchi AH, Nadri S. TRAILexpressing recombinant Lactococcus lactis induces apoptosis in human colon adenocarcinoma SW 480 and HCT 116 cells. Journal of applied microbiology 2019; 126(5): 1558-1567. [DOI:10.1111/jam.14237]
22. Perdigón G, Fuller R, Raya R. Lactic acid bacteria and their effect on the immune system. Current issues in intestinal microbiology 2001; 2(1): 27-42.
23. Wells J, Robinson K, Chamberlain L, Schofield K, Le Page R. Lactic acid bacteria as vaccine delivery vehicles. Antonie Van Leeuwenhoek 1996; 70(2-4): 317-330. [DOI:10.1007/BF00395939]
24. Ramasamy R, Yasawardena S, Zomer A, Venema G, Kok J, Leenhouts K. Immunogenicity of a malaria parasite antigen displayed by Lactococcus lactis in oral immunisations. Vaccine 2006; 24(18): 3900-3908. [DOI:10.1016/j.vaccine.2006.02.040]
25. Lim S, Jahanshiri F, Abdul Rahim R, Sekawi Z, Yusoff K. Surface display of respiratory syncytial virus glycoproteins in Lactococcus lactis NZ9000. Letters in applied microbioloogy 2010; 51(6): 658-664. [DOI:10.1111/j.1472-765X.2010.02950.x]
26. Cortes-Perez NG, Azevedo V, Alcocer-González JM, Rodriguez-Padilla C, Tamez-Guerra RS, Corthier G, Gruss A, Langella P, Bermúdez-Humarán LG. Cell-surface display of E7 antigen from human papillomavirus type-16 in Lactococcus lactis and in Lactobacillus plantarum using a new cell-wall anchor from lactobacilli. Journal of drug targeting 2005; 13(2): 89-98. [DOI:10.1080/10611860400024219]
27. Kuipers OP, de Ruyter PG, Kleerebezem M, de Vos WM. Quorum sensing-controlled gene expression in lactic acid bacteria. Journal of biotechnology 1998; 64(1): 15-21. [DOI:10.1016/S0168-1656(98)00100-X]
28. Wegkamp A, van Oorschot W, de Vos WM, Smid EJ. Characterization of the role of para-aminobenzoic acid biosynthesis in folate production by Lactococcus lactis. Appllied and environmental microbiology 2007; 73(8): 2673-2681. [DOI:10.1128/AEM.02174-06]
29. Zhang HX, Qiu YY, Zhao YH, Liu XT, Liu M, Yu AL. Immunogenicity of oral vaccination with Lactococcus lactis derived vaccine candidate antigen (UreB) of Helicobacter pylori fused with the human interleukin 2 as adjuvant. Molecular and cellular probes 2014; 28(1): 25-30. [DOI:10.1016/j.mcp.2013.08.003]
30. Perez C, Eichwald C, Burrone O, De Mendoza D. Rotavirus vp7 antigen produced by Lactococcus lactis induces neutralizing antibodies in mice. Journal of applied microbiology 2005; 99(5): 1158-1164. [DOI:10.1111/j.1365-2672.2005.02709.x]
31. Maruyama M, Abe R, Shimono T, Iwabuchi N, Abe F, Xiao JZ. The effects of non-viable Lactobacillus on immune function in the elderly: a randomised, double-blind, placebo-controlled study. International journal of food sciences nutrition 2016; 67(1): 67-73. [DOI:10.3109/09637486.2015.1126564]
32. Ting WJ, Kuo WW, Hsieh DJY, Yeh YL, Day CH, Chen YH, Chen RJ, Padma VV, Chen YH, Huang CY. Heat killed Lactobacillus reuteri GMNL-263 reduces fibrosis effects on the liver and heart in high fat diet-hamsters via TGF-β suppression. International journal of molecular sciences 2015; 16(10): 25881-25896. [DOI:10.3390/ijms161025881]
33. Torii S, Torii A, Itoh K, Urisu A, Terada A, Fujisawa T, Yamada K, Suzuki H, Ishida Y, Nakamura F, Kanzato H, Sawada D, Nonaka A, Hatanaka M, Fujiwara S. Effects of oral administration of Lactobacillus acidophilus L-92 on the symptoms and serum markers of atopic dermatitis in children. International archives of allergy and immunology 2011; 154(3): 236-245. [DOI:10.1159/000321110]
34. Bermúdez-Humarán LG, Cortes-Perez NG, Le Loir Y, Alcocer-González JM, Tamez-Guerra RS, de Oca-Luna RM, Langella P. An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci. Journal of medical microbiology 2004; 53(5): 427-433. [DOI:10.1099/jmm.0.05472-0]
35. Leclerc C, Charbit A, Molla A, Hofnung M. Antibody response to a foreign epitope expressed at the surface of recombinant bacteria: importance of the route of immunization. Vaccine 1989; 7(3): 242-248. [DOI:10.1016/0264-410X(89)90237-5]
36. Georgiou G, Stathopoulos C, Daugherty PS, Nayak AR, Iverson BL, Curtiss III R. Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines. Nature biotechnology 1997; 15(1): 29-34. [DOI:10.1038/nbt0197-29]
37. Mazza G, Dunne D, Butterworth A. Antibody isotype responses to the Schistosoma mansoni schistosomulum in the CBA/N mouse induced by different stages of the parasite life cycle. Parasite immunology 1990; 12(4‐5): 529-543. [DOI:10.1111/j.1365-3024.1990.tb00986.x]
38. Atmadja AK, Atkinson R, Sartono E, Partono F, Yazdanbakhsh M, Maizels RM. Differential decline in filaria-specific IgG1, IgG4, and IgE antibodies in Brugia malayi-infected patients after diethylcarbamazine chemotherapy. Journal of infection disease 1995; 172(6): 1567-1572. [DOI:10.1093/infdis/172.6.1567]
39. Al-Qaoud KM, Abdel-Hafez SK. Humoral and cytokine response during protection of mice against secondary hydatidosis caused by Echinococcus granulosus. Parasitology research 2005; 98(1): 54-60. [DOI:10.1007/s00436-005-0004-z]
40. Yong W, Heath D, Van Knapen F. Comparison of cestode antigens in an enzyme-linked immunosorbent assay for the diagnosis of Echinococcus granulosus, Taenia hydatigena and T ovis infections. in sheep. Research in veterinary sciences 1984; 36(1): 24-31. [DOI:10.1016/S0034-5288(18)31996-9]
41. Zhang W, You H, Li J, Zhang Z, Turson G, Aili H, Wang J, Mcmanus DP. Immunoglobulin profiles in a murine intermediate host model of resistance for Echinococcus granulosus infection. Parasite immunology 2003; 25(3): 161-168. [DOI:10.1046/j.1365-3024.2003.00622.x]
42. Woollard D, Gauci C, Heath D, Lightowlers M. Epitope specificities and antibody responses to the EG95 hydatid vaccine. Parasite immunology 1998; 20(11): 535-540. [DOI:10.1046/j.1365-3024.1998.00176.x]
43. Lin R, Ding J, Lu X, Wang X, Wei X, Wang Y, Wen H. Transient expression of Echinococcus granulosus Eg95 DNA vaccine and induction of immune response in mice. Chinese journal of parasitology & parasitic diseases 2004; 22(4): 204-208.
44. Heath D, Jensen O, Lightowlers M. Progress in control of hydatidosis using vaccination-a review of formulation and delivery of the vaccine and recommendations for practical use in control programmes. Acta tropica 2003; 85(2): 133-143. [DOI:10.1016/S0001-706X(02)00219-X]
45. Miquel‐Clopés A, Bentley E, Stewart J, Carding S. Mucosal vaccines and technology. Clinical and experimental immunology 2019; 196(2): 205-214. [DOI:10.1111/cei.13285]
46. Kak G, Raza M, Tiwari BK. Interferon-gamma (IFN-γ): exploring its implications in infectious diseases. Biomolecular concepts 2018; 9(1): 64-79. [DOI:10.1515/bmc-2018-0007]
47. Rigano R, Profumo E, Di Felice G, Ortona E, Teggi A, Siracusano A. In vitro production of cytokines by peripheral blood mononuclear cells from hydatid patients. Clinical and experimental immunology 1995; 99(3): 433-439. [DOI:10.1111/j.1365-2249.1995.tb05569.x]
48. Rigano R, Profumo E, Ioppolo S, Notargiacomo S, Teggi A, Siracusano A. Cytokine patterns in seropositive and seronegative patients with Echinococcus granulosus infection. Immunology letters 1998; 64(1): 5-8. [DOI:10.1016/S0165-2478(98)00072-8]
49. Mezioug D, Touil-Boukoffa C. Cytokine profile in human hydatidosis: possible role in the immunosurveillance of patients infected with Echinococcus granulosus. Parasite 2009; 16(1): 57-64. [DOI:10.1051/parasite/2009161057]
50. Ding JB, Ma XM, Wei XL, Jia HY, Mamuti W, Lin RY, Hou M, Wen H. Th1 and Th2 cytokines in mice infected with Echinococcus granulosus and immunized with Eg95 genetic vaccine. Cell biology international 2008; 32(3): S48-S49. [DOI:10.1016/j.cellbi.2008.01.208]

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