Mycobacterium tuberculosis short mutant H37Rv-S with reduced growth adaptability is more readily recognized by the host immune system

Jilin Dong; Guilian Li; Jiahao Wang; Bingbing Liu; Yu Xiang; Songhao Jiang; Jiahui Shi; Wenhui Wu; Guibin Wang; Lei Chang; Chen Wu; Haican Liu; Ping Xu; Yao Zhang

doi:10.1016/j.micpath.2024.107128

Mycobacterium tuberculosis short mutant H37Rv-S with reduced growth adaptability is more readily recognized by the host immune system

Microb Pathog. 2024 Nov 18:107128. doi: 10.1016/j.micpath.2024.107128. Online ahead of print.

Authors

Jilin Dong¹, Guilian Li², Jiahao Wang³, Bingbing Liu⁴, Yu Xiang², Songhao Jiang⁵, Jiahui Shi⁵, Wenhui Wu⁶, Guibin Wang⁵, Lei Chang⁵, Chen Wu⁷, Haican Liu⁸, Ping Xu⁹, Yao Zhang¹⁰

Affiliations

¹ College of Life Sciences, Hebei University, 071002 Baoding, China; State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, 102206 Beijing, China.
² National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 102206 Beijing, China.
³ School of Basic Medical Science, Anhui Medical University, 230032 Hefei, China.
⁴ Henan Key Laboratory of Industrial Microbial Resources and Fermentation Technology, College of Biological and Chemical Engineering, Nanyang Institute of Technology, 473004 Nanyang, China.
⁵ State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, 102206 Beijing, China.
⁶ State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, 102206 Beijing, China; State Key Laboratory of Dampness Syndrome of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Second Clinical Medicine Collage, Guangzhou Higher Education Mega Center, Guangzhou University of Chinese Medicine, 510006 Guangzhou, China.
⁷ College of Life Sciences, Hebei University, 071002 Baoding, China. Electronic address: dawnwuchen@163.com.
⁸ National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 102206 Beijing, China. Electronic address: liuhaican@icdc.cn163.com.
⁹ College of Life Sciences, Hebei University, 071002 Baoding, China; State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, 102206 Beijing, China; School of Basic Medical Science, Anhui Medical University, 230032 Hefei, China; State Key Laboratory of Dampness Syndrome of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Second Clinical Medicine Collage, Guangzhou Higher Education Mega Center, Guangzhou University of Chinese Medicine, 510006 Guangzhou, China. Electronic address: xuping_bprc@126.com.
¹⁰ State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Beijing Institute of Lifeomics, 102206 Beijing, China. Electronic address: zhangyaowsw@163.com.

PMID: 39566829
DOI: 10.1016/j.micpath.2024.107128

Abstract

Mycobacterium tuberculosis (Mtb) is the bacterium responsible for causing Tuberculosis (TB) and understanding its mechanisms of virulence, persistence, and pathogenesis is a global research priority. Attenuated strains of Mtb are valuable tools for investigating the genes and proteins involved in these processes. In this study, we identified an Mtb mutant, H37Rv-S, which exhibits a shorter mycelium, smoother colony, slower growth, and reduced antibiotics resistance compared to the wild-type strain H37Rv. Genomic sequencing revealed 34 mutation events in the coding regions of H37Rv-S, affecting 31 genes. We conducted TMT-labeling quantitative proteomics to compare the expression differences between H37Rv-S and H37Rv, as well as their infected bone marrow derived macrophages (BMDMs). The results showed that 716 protein groups (23.96%) in H37Rv-S and 115 protein groups (2.99%) in the infected BMDMs were differentially expressed. The dysregulated proteins in H37Rv-S correspond with its phenotype characteristics. Among the 31 affected genes in H37Rv-S, 10 showed upregulation and 1 showed downregulation at the protein level. Notable, 16 associated network proteins in the phoP/phoR system were significantly dysregulated due to a frameshift mutation in phoP, altering its protein sequence after Phe45. The dysregulated host proteins in H37Rv-S were associated with immune response, necroptosis, and ferroptosis. Additionally, H37Rv-S demonstrated reduced survival capability in strain fluorescence labeling and colony-forming unit (CFU) counting post-infection of BMDM cells. These findings suggest that H37Rv-S is an attenuated strain exhibiting defective phenotype characteristics and is more readily recognized and eliminated by the host. This enhanced understanding of the differences between virulent and attenuated strains could facilitate the development of new targets and therapeutics for TB prevention and treatment.

Keywords: Mycobacterium tuberculosis; growth; macrophage; mutant; quantitative proteome.