Comparative analysis of mce operon gene expression levels of Mycobacterium tuberculosis during infection of human and bovine macrophages

• Grant number: 17/24903-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2018
• End date: June 16, 2019
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Ana Marcia de Sá Guimarães
• Grantee: Felipe Silva
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 18/18674-1 - Comparative lipid profiling of a mce3 mutant of Mycobacterium tuberculosis and M. tuberculosis complex wild type strains, BE.EP.IC

Abstract

Tuberculosis is an important disease caused by members of the Mycobacterium tuberculosis Complex (MTBC) that affect human beings and animals. These bacterial are described by clonal evolutionary processes from a Mycobacterium tuberculosis ancestral, demonstrating great genomic similarity, but with different phenotypic profiles among host adaption and virulence. The virulence features analysis, such as the MCE (Mammalian Cell Entry) family, differentially present among MTBC species, can be a tool to comprehend interactions of these pathogens with host infected cells, and also to understand their host adaptability. Thus, this project aims to evaluate genic expression levels of the mce1 to 4 operons in different MTBC strains (M. tuberculosis H37Rv, M. tuberculosis CDC1551, M. tuberculosis CDC1551 mce3A-, e Mycobacterium bovis SP38) in different times in vitro post infection in human and bovine macrophages. These M. tuberculosis and M. bovis strains will be maintained and pre-quantified in 7H9-OAD medium, and will be selected for the macrophages (human THP-1 and peripheral blood monocyte-derived macrophages) infections. Total RNA will be extracted from the infected cells in different times, using Trizol and commercial kit containing DNase, followed by cDNA construction through RT-PCR. The expression of the 4 MCE families will be verified by qPCR and the expression level will be statistically compared by Ct method. The results will serve as data to comparative analysis of the infection dynamic of these MTBC species in human and bovine macrophages, and will consist the first step to fitness differentiation identification among these microorganisms in macrophages from different host species. (AU)