The effect of oral administration Lactococcus lactis probiotic vaccine carrying Brucella abortus BLS antigen on changes in blood leukocyte ratio and CRP as biomarkers in brucellosis

authors:

avatar Fatehi Fatehi , avatar Abbas Doosti ORCID , * , avatar Mohammad Saeid Jami


how to cite: Fatehi F, Doosti A, Jami M S. The effect of oral administration Lactococcus lactis probiotic vaccine carrying Brucella abortus BLS antigen on changes in blood leukocyte ratio and CRP as biomarkers in brucellosis. koomesh. 2023;25(6):e152862. 

Abstract

Introduction: Oral vaccines in infectious diseases, whose agents enter the body through the mucous membrane, will induce the innate immune pathway. The neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been documented as biomarkers of Brucella systemic inflammation. The purpose of this study is to investigate the effect of oral administration of the Lactococcus lactis probiotic vaccine carrying Brucella abortus BLS antigen on the ratio of blood leukocytes as biological biomarkers in mice with brucellosis compared to the control group. Materials and Methods: 60 mice with brucellosis (challenged) and 60 healthy mice (non-challenged) were included in the study. The challenge group was divided into 6 subgroups and 6 types of intervention including 1) L. lactis/pNZ8148-Usp45-BLS, 2) L. lactis-pNZ8148, 3) L. lactis, 4) IRIBA Vac, 5) pNZ8148 and 6) PBS received by gastric gavage in all groups except IRIBA Vac. Serum was used to check hematological parameters. C-reactive protein (CRP) level was measured. The number of neutrophils and lymphocytes was determined using a complete blood cell count. PLR and NLR were calculated. Results: In this study, the level of CRP in the mice of the challenge group increased significantly compared to the non-challenge group. (P<0.0001) The mice receiving oral vaccine L. lactis/pNZ8148-Usp45-BLS were able to inhibit the infection and reduce the concentration of CRP in They were basic level. Mice with brucellosis have lower levels of NLR and PLR than the control group, and after receiving the recombinant vaccine, an increase in leukocytes was observed (P˂0.0001). Conclusion: The mice of the challenge group and the non-challenge group showed a statistically significant difference in terms of PLR and NLR as well as CRP. These parameters can be useful markers in evaluating the systemic inflammation of brucellosis.

References

  • 1.

    Sing A, editor. Zoonoses-Infections affecting humans and animals: Focus on public health aspects. Springer; 2014 Dec 8.##https://doi.org/10.1007/978-94-017-9457-2.

  • 2.

    Amjadi O, Rafiei A, Mardani M, Zafari P, Zarifian A. A review of the immunopathogenesis of Brucellosis. Infect Dis 2019; 51: 321-333.

  • 3.

    Kurmanov B, Zincke D, Su W, Hadfield TL, Aikimbayev A, Karibayev T, et al. Assays for identification and differentiation of Brucella species: A review. Microorganisms 2022; 10: 1584.

  • 4.

    Burn GL, Foti A, Marsman G, Patel DF, Zychlinsky A. The neutrophil. Immunity 2021; 54: 1377-1391.

  • 5.

    Rezaei M, Rabbani-Khorasgani M, Zarkesh-Esfahani SH, Emamzadeh R, Abtahi H. Prediction of the Omp16 Epitopes for the development of an Epitope-based vaccine against Brucellosis. Infect Disord Drug Targets 2019; 19: 36-45.

  • 6.

    Ribeiro LA, Azevedo V, Le Loir Y, Oliveira SC, Dieye Y, Piard JC, Gruss A, Langella P. Production and targeting of the Brucella abortus antigen L7/L12 in Lactococcus lactis: a first step towards food-grade live vaccines against brucellosis. Appl Environ Microbiol 2002; 68: 910-916.

  • 7.

    Badmasti F, Habibi M, Firoozeh F, Fereshteh S, Bolourchi N, Goodarzi NN. The combination of CipA and PBP-7/8 proteins contribute to the survival of C57BL/6 mice from sepsis of Acinetobacter baumannii. Microb Pathog 2021; 158: 105063.

  • 8.

    Liu X, Qi L, Lv J, Zhang Z, Zhou P, Ma Z, et al. The immune response to a recombinant Lactococcus lactis oral vaccine against foot-and-mouth disease virus in mice. Biotechnol Lett 2020; 42: 1907-1917.

  • 9.

    Bermdez-Humarn LG, Langella P, Cortes-Perez NG, Gruss A, Tamez-Guerra RS, Oliveira SC, et al. Intranasal immunization with recombinant Lactococcus lactis secreting murine interleukin-12 enhances antigen-specific Th1 cytokine production. Infect Immun 2003; 71: 1887-1896.

  • 10.

    O'callaghan D. Human brucellosis: recent advances and future challenges. Infect Dis Poverty 2020; 9: 1-2.

  • 11.

    Blasco JM. Control and eradication strategies for Brucella melitensis infection in sheep and goats. Prilozi 2010; 31: 145-165.

  • 12.

    Dorneles E, Sriranganathan N, Lage AP. Recent advances in Brucella abortus vaccines. Vet Res 2015; 46: 76.

  • 13.

    Vishnu US, Sankarasubramanian J, Gunasekaran P, Rajendhran J. Novel vaccine candidates against Brucella melitensis identified through reverse vaccinology approach. Omics 2015; 19: 722-729.

  • 14.

    D'Souza R, Pandeya DR, Hong ST. Lactococcus lactis: an efficient Gram positive cell factory for the production and secretion of recombinant protein. Biomed Res 2012; 23: 1-7.

  • 15.

    Rezaei M, Rabbani-Khorasgani M, Zarkesh-Esfahani SH, Emamzadeh R, Abtahi H. Lactococcus-based vaccine against brucellosis: IgG immune response in mice with rOmp16-IL2 fusion protein. Arch Microbiol 2021; 203: 2591-2596.

  • 16.

    Akbari R, Sekhavati MH, Bahrami A, Majidzadeh Heravi R, Yousefi S. Production of brucella lumazine synthase recombinant protein to design a subunit vaccine against undulant fever. Arch Razi Inst 2019; 74: 1-6.

  • 17.

    Tirbakhsh Gouran S, Doosti A, Jami MS. Expression of brucella abortus Omp25 protein in lactococcus lactis probiotic bacteria. J Mazandaran Univ Med Sci 2023; 32: 16-31.

  • 18.

    Moreno E, Barquero-Calvo E. The role of neutrophils in brucellosis. Microbiol Mol Biol Rev 2020; 84: e00048-20.