Recombinant fusion protein LipL41-OmpL1 as a Potential Candidate for a Cost-Effective Vaccine against Leptospirosis

Document Type : Original Articles

Authors

1 Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran

2 Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

3 Department of Microbiology, falavarjan Branch, Islamic Azad University, Isfahan, Iran

4 Department of Human vaccine , Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

5 Department of Immunology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

10.32592/ARI.2024.79.5.1099

Abstract

Leptospirosis is a disease that threatens the health of humans and animals, which is caused by pathogenic Leptospira. Early detection of pathogenic Leptospires prevents many problems. LipL41-OmpL1, a protected outer membrane protein (OMP) of pathogenic Leptospira, was inserted into E. coli bacteria using different software for the amino acid sequence of OmpL1 and LipL41 to design a recombinant fusion protein and then expressed to investigate immunogenicity. The selected genes were propagated and cloned as a fusion in pET32a+ plasmid vector and expressed by Escherichia coli plays S (DE3) by heat shock method. It was evaluated in the BALB/c mouse laboratory animal model. Recombinant LipL41-OmpL1 protein was confirmed using urea purification method and western blot and its immunogenicity was evaluated by measuring high humoral immune stimulation and antibody secretion in BALB/c mice by ELISA method. The results showed that animals that received both OmpL1 and LipL41 proteins had 85% immunogenicity, while the control group that did not receive the fusion protein had only 25% immunogenicity (P < 0.001). Furthermore, no evidence of infection was found in recipients of the OmpL1-LipL41 fusion protein, suggesting that this protein is safe to use. The protective effects of immunization with OmpL1 and LipL41 were synergistic, as significant levels of protection were not observed in animals immunized with OmpL1 or LipL41 alone. Overall, this study highlights the potential of recombinant OmpL1 and LipL41 fusion protein as a promising research strategy on the development of vaccines and ELISA diagnostic kit for prevention and rapid diagnosis of leptospirosis.

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Main Subjects

References

  1. Philip N, Bahtiar Affendy N, Ramli SNA, Arif M, Raja P, Nagandran E, et al. Leptospira interrogans and Leptospira kirschneri are the dominant Leptospira species causing human leptospirosis in Central Malaysia. PLoS neglected tropical diseases. 2020;14(3):e0008197.
  2. Evangelista K, Franco R, Schwab A, Coburn J. Leptospira interrogans binds to cadherins. PLoS Neglected Tropical Diseases. 2014;8(1):e2672.
  3. Adler B, de la Peña Moctezuma A. Leptospira and leptospirosis. Veterinary microbiology. 2010;140(3-4):287-96.
  4. Adler B. Vaccines against leptospirosis. Leptospira and leptospirosis. 2014:251-72.
  5. Cerqueira GM, Picardeau M. A century of Leptospira strain typing. Infection, Genetics and Evolution. 2009;9(5):760-8.
  6. Nally JE, Whitelegge JP, Bassilian S, Blanco DR, Lovett MA. Characterization of the outer membrane proteome of Leptospira interrogans expressed during acute lethal infection. Infection and immunity. 2007;75(2):766-73.
  7. Nisa S, Wilkinson DA, Angelin-Bonnet O, Paine S, Cullen K, Wright J, et al. Diverse epidemiology of Leptospira serovars notified in New Zealand, 1999–2017. Pathogens. 2020;9(10):841.
  8. Thongboonkerd V, Chiangjong W, Saetun P, Sinchaikul S, Chen ST, Kositanont U. Analysis of differential proteomes in pathogenic and non‐pathogenic Leptospira: Potential pathogenic and virulence factors. Proteomics. 2009;9(13):3522-34.
  9. Soltani M, Khaki P, Moradi Bidhendi S, Shahhosseiny M. Molecular characterization of the lipL41 gene of Leptospira interrogans vaccinal serovars in Iran. Archives of Razi Institute. 2015;70(3):145-50.
  10. Holt SC. Anatomy and chemistry of spirochetes. Microbiological reviews. 1978;42(1):114-60.
  11. Cullen PA, Cordwell SJ, Bulach DM, Haake DA, Adler B. Global analysis of outer membrane proteins from Leptospira interrogans serovar Lai. Infection and immunity. 2002;70(5):2311-8.
  12. Haake DA, Levett PN. Leptospirosis in humans. Leptospira and leptospirosis. 2015:65-97.
  13. De Abreu Fonseca C, Teixeira MMG, Romero EC, Tengan FM, Da Silva MV, Shikanai-Yasuda MA. Leptospira DNA detection for the diagnosis of human leptospirosis. Journal of Infection. 2006;52(1):15-22.
  14. Klaasen H, Molkenboer M, Vrijenhoek M, Kaashoek M. Duration of immunity in dogs vaccinated against leptospirosis with a bivalent inactivated vaccine. Veterinary microbiology. 2003;95(1-2):121-32.
  15. Haake DA, Matsunaga J. Characterization of the leptospiral outer membrane and description of three novel leptospiral membrane proteins. Infection and immunity. 2002;70(9):4936-45.
  16. Senthilkumar TMA, Subathra M, Ramadass P. Evaluation of recombinant leptospiral antigen LipL41 in enzymelinked immunosorbent assay and latex agglutination test for serodiagnosis of canine leptospirosis. Veterinarski arhiv. 2007;77(6):475-84.
  17. Hoseinpur R, Khaki P, Noofeli M, Moradi Bidhendi S. Molecular detection of pathogenic leptospiral serovars by PCR, based on lipL21 gene. Archives of Razi Institute. 2015;70(4):223-7.
  18. Golab N, Khaki P, Harzandi N, Esmaelizad M, Tebianian M. Ekspresija i pročišćavanje površinskog lipoproteinskog antigena Lipl41 iz patogenih bakterija roda Leptospira. Veterinarski arhiv. 2020;90(3):297-305.
  19. Patricia H-R, Arlen GR, Mónica B, Gladys Q. Identification of ompL1 and lipL32 Genes to Diagnosis of Pathogenic Leptospira spp. isolated from Cattle. Open Journal of Veterinary Medicine. 2014;2014.
  20. Raja V, Natarajaseenivasan K. Pathogenic, diagnostic and vaccine potential of leptospiral outer membrane proteins (OMPs). Critical reviews in microbiology. 2015;41(1):1-17.
  21. Sakamoto M, Kato T, Sato F, Yoshikawa K, Yoshida M, Shiba K, et al. A case of leptospirosis caused by Leptospira borgpetersenii serovar sejroe infected in Bali Island, Indonesia. Kansenshogaku zasshi The Journal of the Japanese Association for Infectious Diseases. 2005;79(4):294-8.
  22. Costa F, Wunder Jr EA, De Oliveira D, Bisht V, Rodrigues G, Reis MG, et al. Patterns in Leptospira shedding in Norway rats (Rattus norvegicus) from Brazilian slum communities at high risk of disease transmission. PLoS neglected tropical diseases. 2015;9(6):e0003819.
  23. Golab N, Khaki P, Harzandi N, Esmaelizad M, Tebianian M. Expression and purification of the Lipl41, a surface-exposed lipoprotein antigen of pathogenic Leptospira spp. Veterinarski arhiv. 2020;90(3):297-305.
  24. Khaki P, Moradi Bidhendi S, Chang Y-F, Soltani MS, Tadaion K. Amplification and cloning of a gene encoding a 41 kDa outer membrane protein (LipL41) of Leptospira interrogans serovar Canicola. Infection Epidemiology and Microbiology. 2016;2(3):5-7.
  25. Kodjo A, Calleja C, Loenser M, Lin D, Lizer J. A rapid in-clinic test detects acute leptospirosis in dogs with high sensitivity and specificity. Biomed research international. 2016;2016.
  26. Lin M-H, Chang Y-C, Hsiao C-D, Huang S-H, Wang M-S, Ko Y-C, et al. LipL41, a hemin binding protein from Leptospira santarosai serovar Shermani. PLoS One. 2013;8(12):e83246.
  27. Haji Hajikolaei M., Rezaei S, Ghadrdan Mashhadi A, Ghorbanpour M, Abdollahpour G. Comparison of C interrogans infection in the goats and sheep. Iranian Journal of Veterinary Medicine, 2016;10(2):113-119.
  28. Maleki S, Abdollahpour G, Bahonar A. Serological and bacteriological study of leptospirosis in dairy herds and feedlot in Tehran suburbs. Iranian Journal of Veterinary Medicine, 2013; 7(3): 177-183.
  29. Abdollahpour G, Shafighi S-T, Sattari TAbrizi S. Serodiagnosis of leptospirosis in cattle in north of Iran, Gilan. Iranian Journal of Veterinary Medicine, 2009; 3(1): 5-6.
Archives of Razi Institute
Volume 79, Issue 5
September and October 2024
Pages 1099-1108

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