In-Vitro Potential of Recombinant Omp31 Protein from Brucella Melitensis in Inducing the Proliferation of Immunized Mouse Lymphocytes

Document Type : Original Article (s)

Authors

1 Assistant Professor, Department of Microbiology and Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Professor, Department of Microbiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

3 Associate Professor, Nanobiotechnology Research Center, Avecina Research Institute AND Immunology Research Center, Tehran University of Medical Sciences, Tehran, Iran

4 Professor, Monoclonal Antibody Research Center, Avicenna Research Institute, The Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

5 Associate Professor, Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

Abstract

Background: Brucella melitensis infection is still a major health problem for human and cattle in developing countries and the Middle East. Cell-mediated immunity is the dominant immune response required for protection against brucellosis. So, identifying of new candidates which can induce cell-mediated immunity is demanded. Proliferation assay is one of the methods used to evaluate the potential of an antigen to generate memory T-cell. In the present study, the proliferation of speloncyets obtained from immunized mice was evaluated when they were challenged by recombinant Omp31 (rOmp31) in vitro.Methods: Mice were immunized by phosphate buffered saline (PBS), rOmp31 and formalin-killed Brucella melitensis Rev.1 with Freund’s adjuvant. At the day 30, after the last immunization, spleens were removed and homogenized. Purified rOmp31 (2.5 and 5 µg/ml) was added to 2 × 105 spleen cells and then, the speloncytes were incubated at 37◦C in 5% CO2 atmosphere. After 48 hours, XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays were used to show the splenocyte proliferation.Findings: The splenocytes of mice immunized with rOmp31or Rev.1 started to proliferate after stimulation with rOmp31. There was a significant difference between SIs obtained from rOmp31 and Rev compared to phosphate buffered saline group (P < 0.01 and P < 0.05, respectively).Conclusion: Based on our findings, rOmp31 could generate a good memory cellular immune response. In addition, the method described here to study the speloncyte proliferation using XTT, is a simple and efficient method.

Keywords

References

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Journal of Isfahan Medical School
Volume 32, Issue 292 - Serial Number 292
May and June 2014
Pages 1036-1045

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History

  • Receive Date: 16 November 2013
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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