Immunization against Leishmania major infection in BALB/c mice using a subunit-based DNA vaccine derived from TSA, LmSTI1, KMP11, and LACK predominant antigens

Document Type : Original Article

Authors

1 Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

3 Department of Medical Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

5 Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran

6 Centre for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran

7 Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

8 Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

9 Department of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): To design a multivalent DNA vaccine encoding the most immunogenic regions of the Leishmania major antigens including TSA (Thiol-specific antioxidant protein), LmSTI1 (Leishmania major stress-inducible protein1), LACK (Leishmania homologue of receptors for activated C Kinase), and KMP11 (kinetoplastid membrane protein-11) on BALB/c mice.
Materials and Methods: The chimeric construct was generated including the most immunogenic epitopes containing a combination of domains and oligopeptides of the aforementioned genes. The construct was cloned into pcDNA 3.1 plasmid and named “pleish-dom.” Following intramuscular injection of mice, the capability of the vector pleish-dom alone and with pIL-12 (expressing murine IL-12) to raise protective cytokines and parasite burden was evaluated in the BALB/c mice as a susceptible animal model against L. major.
Results: The immunized mice with pleish-dom/pIL-12 showed the highest and the lowest levels of interferon-gamma (IFN-γ) and interleukin-10 (IL-10), as well as the lowest leishmanin skin test (LST) reactions, were found through 8 weeks post-infection.
Conclusion: Although the obtained DNA vaccine from the immunogenic chimeric protein of L. major antigens was able to induce a high level of IFN-γ, it partially protected mice against L. major. However co-administration with pIL-12 led to shift immune response to Th1 phenotype, granuloma formation, and lowered parasite burden, and consequently distinct protection was found.

Keywords

References

1. O mondiale de la Santé, World Health Organization. Leishmaniasis in high-burden countries: an epidemiological update based on data reported in 2014. Wkly Epidemiol Rec 2016; 91:287-296.
2. Leishmaniases WECotCot, Meeting WECotCotL, World Health Organization. Control of the Leishmaniases: Report of a WHO Expert Committee: World Health Organization 1990.
3. Khanjani N, González U, Leonardi‐Bee J, Mohebali M, Saffari M, Khamesipour A. Vaccines for preventing cutaneous leishmaniasis. Cochrane Database Syst Rev 2009.
4. Nadim A,Javadian E, Mohebali M. The experience of leishmanization in the Islamic Republic of Iran. East Mediterr Health J 1997; 3:284-290.
5. Mohebali M, Khamesipour A, Mobedi I, Zarei Z, Hashemi-Fesharki R. Double-blind randomized efficacy field trial of alum precipitated autoclaved Leishmania major vaccine mixed with BCG against canine visceral leishmaniasis in Meshkin-Shahr district, IR Iran. Vaccine 2004; 22:4097-4100.
6. Bogdan C, Schröppel K, Lohoff M, Röllinghoff M, Solbach W. Immunization of susceptible hosts with a soluble antigen fraction from Leishmania major leads to aggravation of murine leishmaniasis mediated by CD4+ T cells. Eur J Immunol 1990; 20:2533-2540.
7. Ramírez L, Iborra S, Cortés J, Bonay P, Alonso C, Barral-Netto M, et al. BALB/c mice vaccinated with Leishmania major ribosomal proteins extracts combined with CpG oligodeoxynucleotides become resistant to disease caused by a secondary parasite challenge. J Biomed Biotecnol 2010;2010:e181690.
8. Dey R, Dagur PK, Selvapandiyan A, McCoy JP, Salotra P, Duncan R, et al. Live attenuated Leishmania donovani p27 gene knockout parasites are nonpathogenic and elicit long-term protective immunity in BALB/c mice. J Immunol 2013; 190:2138-2149.
9. Fiuza JA, Dey R, Davenport D, Abdeladhim M, Meneses C, Oliveira F, et al. Intradermal immunization of Leishmania donovani centrin knock-out parasites in combination with salivary protein LJM19 from sand fly vector induces a durable protective immune response in hamsters. PLoS Negl Trop Dis 2016; 10:e0004322.
10. Coler RN, Goto Y, Bogatzki L, Raman V, Reed SG. Leish-111f, a recombinant polyprotein vaccine that protects against visceral Leishmaniasis by elicitation of CD4+ T cells. Infect Immun 2007; 75:4648-4654.
11. Gillespie PM, Beaumier CM, Strych U, Hayward T, Hotez PJ, Bottazzi ME. Status of vaccine research and development of vaccines for leishmaniasis. Vaccine 2016; 34:2992-2995.
12. Ahmed SBH, Touihri L, Chtourou Y, Dellagi K, Bahloul C. DNA based vaccination with a cocktail of plasmids encoding immunodominant Leishmania (Leishmania) major antigens confers full protection in BALB/c mice. Vaccine 2009; 27:99-106.
13. Leitner WW, Thalhamer J. DNA vaccines for non-infectious diseases: new treatments for tumour and allergy. Expert Opin Biol Th 2003; 3:627-638.
14. Nogueira F, Moreira M, Borja-Cabrera G, Santos F, Menz I, Parra L, et al. Leishmune® vaccine blocks the transmission of canine visceral leishmaniasis: absence of Leishmania parasites in blood, skin and lymph nodes of vaccinated exposed dogs. Vaccine 2005; 23:4805-4810.
15. Regina-Silva S, Feres AM, França-Silva JC, Dias ES, Michalsky ÉM, de Andrade HM, et al. Field randomized trial to evaluate the efficacy of the Leish-Tec® vaccine against canine visceral leishmaniasis in an endemic area of Brazil. Vaccine 2016; 34:2233-2239.
16. Moreno J, Vouldoukis I, Martin V, McGahie D, Cuisinier A-M, Gueguen S. Use of a LiESP/QA-21 vaccine (CaniLeish) stimulates an appropriate Th1-dominated cell-mediated immune response in dogs. PLoS Negl Trop Dis 2012; 6:e1683.
17. Gupta A, Chaphalkar SR. Current approaches and problems in malaria vaccine development. Eur J Biol Res 2016; 6:14-20.
18. Costa CHN, Peters NC, Maruyama SR, de Brito Jr EC, de Miranda Santos IKF, Working Group on Research Priorities for Development of Leishmaniasis Vaccines. Vaccines for the leishmaniases: proposals for a research agenda. PLoS Negl Trop Dis 2011; 5:e943.
19. Ramos I, Alonso A, Marcen J, Peris A, Castillo J, Colmenares M, et al. Heterologous prime-boost vaccination with a non-replicative vaccinia recombinant vector expressing LACK confers protection against canine visceral leishmaniasis with a predominant Th1-specific immune response. Vaccine 2008; 26:333-344.
20. Campos-Neto A, Webb J, Greeson K, Coler R, Skeiky Y, Reed S. Vaccination with plasmid DNA encoding TSA/LmSTI1 leishmanial fusion proteins confers protection against Leishmania major infection in susceptible BALB/c mice. Infect Immun 2002; 70:2828-2836.
21. Shahbazi M, Zahedifard F, Taheri T, Taslimi Y, Jamshidi S, Shirian S, et al. Evaluation of live recombinant nonpathogenic Leishmania tarentolae expressing cysteine proteinase and A2 genes as a candidate vaccine against experimental canine visceral leishmaniasis. PLoS one 2015; 10:e0132794.
22. Rodríguez-Cortés A, Ojeda A, López-Fuertes L, Timón M, Altet L, Solano-Gallego L, et al. Vaccination with plasmid DNA encoding KMPII, TRYP, LACK and GP63 does not protect dogs against Leishmania infantum experimental challenge. Vaccine 2007; 25:7962-7971.
23. Kedzierski L. Leishmaniasis vaccine: where are we today? J Glob Infect Dis. 2010; 2:177-185.
24. Sette A, Fikes J. Epitope-based vaccines: an update on epitope identification, vaccine design and delivery. Curr Opin Immunol 2003; 15:461-470.
25. Ivory C, Chadee K. DNA vaccines: designing strategies against parasitic infections. Genet Vaccines Ther 2004; 2:17.
26. Saade F, Petrovsky N. Technologies for enhanced efficacy of DNA vaccines. Expert Rev Vaccines. 2012; 11:189-209.
27. Okwor I, Uzonna JE. Pathways leading to interleukin-12 production and protective immunity in cutaneous leishmaniasis. Cell Immunol 2016; 309:32-36.
28. Seyed N, Zahedifard F, Safaiyan S, Gholami E, Doustdari F, Azadmanesh K, et al. In silico analysis of six known Leishmania major antigens and in vitro evaluation of specific epitopes eliciting HLA-A2 restricted CD8 T cell response. PLoS Negl Trop Dis 2011; 5:e1295.
29. Agallou M, Athanasiou E, Koutsoni O, Dotsika E, Karagouni E. Experimental validation of multi-epitope peptides including promising MHC class I-and II-restricted epitopes of four known Leishmania infantum proteins. Front Immunol 2014;5: e268.
30. Das S, Freier A, Boussoffara T, Das S, Oswald D, Losch FO,  et al. Modular multiantigen T cell epitope–enriched DNA vaccine against human leishmaniasis. Sci Transl Med 2014; 6:234ra56.
31. Matos  I, Mizenina O, Lubkin A, Steinman RM, Idoyaga J. Targeting Leishmania major antigens to dendritic cells in vivo induces protective immunity. PLoS one 2013; 8: e67453.
32. Jensen K.D, Sercarz EE, Gabaglia CR. Altered peptide ligands can modify the Th2 T cell response to the immunodominant 161-175 peptide of LACK (Leishmania homolog for the receptor of activated C kinase). Mol Immunol 2009; 46:366-374.
33. Azizi H, Kazemi B, Bandehpour M, Mohebali M, Khamesipour A, Aryaeipour M, et al. 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. Iran J Parasitol 2016; 11:480-488.
34. Yadegari Z, Bandehpour M, Kazemi B, Sharifi-Sarasiabi K. Expression of recombinant human amelogenin in iranian lizard Leishmania and its biological function assay. Iran J Public Health 2015; 44:987-996.
35. Buffet P, Sulahian A, Garin Y, Nassar N, Derouin F. Culture microtitration: a sensitive method for quantifying Leishmania infantum in tissues of infected mice. Antimicrob Agents Chemother 1995; 39:2167-2168.
36. Stauber LA. Host resistance to the Khartoum strain of Leishmania donovani. Rice Institute Pamphlet-Rice University Studies 1958; 45-96.
37. Shirzadi M, Esfahania S, Mohebalia M, Ershadia M, Gharachorlo F, Razavia M, et al. Epidemiological status of leishmaniasis in the Islamic Republic of Iran, 1983-2012. East Mediterr Health J 2015; 21:736.
38. Salehi Gh, Fata A, Mohaghegh MA, Mousavi Bazzaz SM, Rafatpanah H, Movahedi A. Molecular identification of Leishmania species in Taybad district, Iran. Asian Pac J Trop Dis 2014; 4:S535-539.
39. Handman E. Leishmaniasis: current status of vaccine development. Clin Microbiol Rev 2001; 14:229-243.
40. Desjeux, P., Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 2004; 27:305-318.
41. Von Stebut E, Tenzer S. Cutaneous leishmaniasis: Distinct functions of dendritic cells and macrophages in the interaction of the host immune system with Leishmania major. Int J Med Microbiol 2018; 308:206-214.
42. Maspi N, Ghaffarifar F, Sharifi Z, Dalimi A, Dayer MS. Comparative assessment of induced immune responses following intramuscular immunization with fusion and cocktail of LeIF, LACK and TSA genes against cutaneous leishmaniasis in BALB/c mice. Arch Immunol Ther Exp 2018; 66:55-64.
43. Coler RN, Reed SG. Second-generation vaccines against leishmaniasis. Trends Parasitol 2005; 21:244-249.
44. Skeiky YA, Coler RN, Brannon M, Stromberg E, Greeson K, Crane RT, et al. Protective efficacy of a tandemly linked, multi-subunit recombinant leishmanial vaccine (Leish-111f) formulated in MPL® adjuvant. Vaccine 2002; 20:3292-3303.
45. Martínez‐López M, Iborra S, Conde‐Garrosa R, Sancho D. Batf3‐dependent CD103+ dendritic cells are major producers of IL‐12 that drive local Th1 immunity against Leishmania major infection in mice. Eur J Immunol 2015; 45:119-129.
46. Darrah PA, Patel DT, De Luca PM, Lindsay R, Davey DF, Flynn BJ, et al. Multifunctional TH1 cells define a correlate of vaccine-mediated protection against Leishmania major. Nat Med 2007; 13:843-850.
47. Teng MW, Bowman EP, McElwee JJ, Smyth MJ, Casanova J-L, Cooper AM, et al. IL-12 and IL-23 cytokines: from discovery to targeted therapies for immune-mediated inflammatory diseases. Nat Med 2015; 21:719.
48. Bhaumik S, Basu R, Sen S, Naskar K, Roy S. KMP-11 DNA immunization significantly protects against L. donovani infection but requires exogenous IL-12 as an adjuvant for comparable protection against L. major. Vaccine 2009; 27:1306-1316.
49. Heinemann C, Heink S, Petermann F, Vasanthakumar A, Rothhammer V, Doorduijn E, et al. IL-27 and IL-12 oppose pro-inflammatory IL-23 in CD4+ T cells by inducing Blimp1. Nat commun 2014; 5:e3770.
50. Cavalcanti AS, Ribeiro-Alves M, de OR Pereira L, Mestre GL, Ferreira ABR, Morgado FN, et al. Parasite load induces progressive spleen architecture breakage and impairs cytokine mRNA expression in Leishmania infantum-naturally infected dogs. PLoS One 2015; 10:e0123009.
51. Abdoli A, Maspi N, Ghaffarifar F. Wound healing in cutaneous leishmaniasis: A double edged sword of IL-10 and TGF-β. Comp Immunol Microbiol Infect Dis 2017; 51:15-26.
52. ScottP, Novais FO. Cutaneous leishmaniasis: immune responses in protection and pathogenesis. Nat Rev Immunol 2016; 16:581-592.
Iranian Journal of Basic Medical Sciences
Volume 22, Issue 12
December 2019
Pages 1493-1501

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