Phylogenetic analysis of prospective <i>M. bovis</i> antigens with the aim of developing candidate vaccines for bovine tuberculosis

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Abstract

Background

Bovine Tuberculosis is a respiratory disease caused by the pathogen Mycobacterium bovis (M. bovis) that infects cattle. Though rare, this disease can also affect humans, as well as domestic and wild animals, making it a serious concern. Therefore, searching for alternative and new vaccines with high efficiency and safety is the main goal in bovine tuberculosis prophylaxis. New vaccines, known as vector vaccines, have the potential to become safe and effective alternatives to the traditional BCG vaccine. In this study, two major immunodominant proteins of M. bovis Esat-6 and TB10.4 were utilized to create a vector vaccine for bovine tuberculosis.

Methods

The Esat-6 and TB10.4 genes were amplified by PCR. The amplified and purified PCR products were sequenced by the Sanger method. Assembly and multiple alignments of amplicon nucleotides were carried out in the MEGA 11 software.

Result

Two genes of the local strain 0078-M. bovis-8/RIBSP were sequenced. The nucleotide sequences were deposited in the GenBank database. Comparative analysis of the nucleotide sequences of the ESAT-6 and TB10.4 genes established 100% identity of the compared strains of Mycobacterium.

Conclusion

Through the use of phylogenetic analysis, it has been confirmed that the amplified genes are related to the mycobacteria genus. This discovery allows the development of a vector vaccine against bovine tuberculosis utilising these genes.

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Most cited references 23

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Loss of RD1 contributed to the attenuation of the live tuberculosis vaccines Mycobacterium bovis BCG and Mycobacterium microti.

Alexander S Pym, Priscille Brodin, Roland Brosch … (2002)

Although large human populations have been safely immunized against tuberculosis with two live vaccines, Mycobacterium bovis BCG or Mycobacterium microti, the vole bacillus, the molecular basis for the avirulence of these vaccine strains remains unknown. Comparative genomics has identified a series of chromosomal deletions common to both virulent and avirulent species but only a single locus, RD1, that has been deleted from M. bovis BCG and M. microti. Restoration of RD1, by gene knock-in, resulted in a marked change in colonial morphology towards that of virulent tubercle bacilli. Three RD1-encoded proteins were localized in the cell wall, and two of them, the immunodominant T-cell antigens ESAT-6 and CFP-10, were also found in culture supernatants. The BCG::RD1 and M. microti::RD1 knock-ins grew more vigorously than controls in immunodeficient mice, inducing extensive splenomegaly and granuloma formation. Increased persistence and partial reversal of attenuation were observed when immunocompetent mice were infected with the BCG::RD1 knock-in, whereas BCG controls were cleared. Knocking-in five other RD loci did not affect the virulence of BCG. This study describes a genetic lesion that contributes to safety and opens new avenues for vaccine development.

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Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guérin attenuation.

Kristine M. Guinn, Vincent Sherman, Graeme L. Hickey … (2003)

The tuberculosis (TB) vaccine bacille Calmette-Guérin (BCG) is a live attenuated organism, but the mutation responsible for its attenuation has never been defined. Recent genetic studies identified a single DNA region of difference, RD1, which is absent in all BCG strains and present in all Mycobacterium tuberculosis (MTB) strains. The 9 open-reading frames predicted within this 9.5-kb region are of unknown function, although they include the TB-specific immunodominant antigens ESAT-6 and CFP-10. In this study, RD1 was deleted from MTB strain H37Rv, and virulence of H37Rv:DeltaRD1 was assessed after infections of the human macrophage-like cell line THP-1, human peripheral blood monocyte-derived macrophages, and C57BL/6 mice. In each of these systems, the H37Rv:DeltaRD1 strain was strikingly less virulent than MTB and was very similar to BCG controls. Therefore, it was concluded that genes within or controlled by RD1 are essential for MTB virulence and that loss of RD1 was important in BCG attenuation.

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Bottlenecks and broomsticks: the molecular evolution of Mycobacterium bovis.

R. Glyn Hewinson, D Clifton, R Rua … (2006)

Mycobacterium bovis is the cause of tuberculosis in cattle and is a member of the Mycobacterium tuberculosis complex. In contrast to many other pathogenic bacterial species, there is little evidence for the transfer and recombination of genes between cells. The clonality of this group of organisms indicates that the population structure is dominated by reductions in diversity, caused either by population bottlenecks or selective sweeps as entire chromosomes become fixed in the population. We describe how these forces have shaped not only the phylogeny of this group but also, at a very local level, the population structure of Mycobacterium bovis in the British Isles. We also discuss the practical implications of applying this knowledge to understanding the spread of infection and the development of improved vaccines and diagnostic tests.

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Author and article information

Contributors

Kamshat Shorayeva: abaizh097@mail.ru

Journal

Journal ID (nlm-ta): J Genet Eng Biotechnol

Journal ID (iso-abbrev): J Genet Eng Biotechnol

Title: Journal of Genetic Engineering & Biotechnology

Publisher: Springer Berlin Heidelberg (Berlin/Heidelberg )

ISSN (Print): 1687-157X

ISSN (Electronic): 2090-5920

Publication date (Electronic): 12 October 2023

Publication date PMC-release: 12 October 2023

Publication date Collection: December 2023

Volume: 21

Electronic Location Identifier: 99

Affiliations

[1 ]GRID grid.466914.8, ISNI 0000 0004 1798 0463, Research Institute for Biological Safety Problems, ; Gvardeysky, 080409 Kazakhstan

[2 ]al-Farabi Kazakh National university, ( https://ror.org/03q0vrn42) Almaty, Kazakhstan

Article

Publisher ID: 552

DOI: 10.1186/s43141-023-00552-3

PMC ID: 10570230

PubMed ID: 37823960

SO-VID: a9b22a7d-7fa8-451f-9add-b2061a994239

License:

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

History

Date received : 28 March 2023

Date accepted : 20 September 2023

Funding

Funded by: FundRef http://dx.doi.org/10.13039/501100004561, Ministry of Education and Science of the Republic of Kazakhstan;

Award ID: IRN AR09259683

Custom metadata

Keywords: bovine tuberculosis,vaccine,m. bovis,esat-6,tb10.4,pcr,sequencing,senger

Data availability:

Keywords: bovine tuberculosis, vaccine, m. bovis, esat-6, tb10.4, pcr, sequencing, senger

Phylogenetic analysis of prospective M. bovis antigens with the aim of developing candidate vaccines for bovine tuberculosis

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Abstract

Background

Methods

Result

Conclusion

Related collections

Vaccines

Most cited references 23

Loss of RD1 contributed to the attenuation of the live tuberculosis vaccines Mycobacterium bovis BCG and Mycobacterium microti.

Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guérin attenuation.

Bottlenecks and broomsticks: the molecular evolution of Mycobacterium bovis.

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