Abstract
Resurgence of mycobacterial diseases particularly tuberculosis has caused a renewed interest to unravel the strategies employed by mycobacteria for intracellular survival. In spite of advancement in mycobacterial research, our knowledge about genes and their corresponding functional proteins involved during the interaction of mycobacterium with host’s macrophages is fragmentary. This study pertains to development of a suitable in vitro model using murine macrophages and Mycobacterium bovis BCG to study proteins expressed during macrophage-myco bacterium interactions. Peritoneal macrophages from BALB/c mice were infected with M. bovis BCG and intracellular replication was assessed by {3H} thymi- dine uptake assay which was maximal when macrophage to mycobacterium ratio was 1:10. SDS-PAGE was employed to study the proteins expressed and selected proteins were subjected to mass spectrometry. Seven proteins found to be upregulated during macrophage-mycobacterium interaction were identified by MALDI-TOF. The results indicate that the present in vitro infection model was able to support the growth of M. bovis BCG in murine macrophages and is an ideal model to determine the pattern of functions of gene expression during the interaction of mycobacterium with macrophages. The differentially expressed proteins will help in understanding the mycobacterial molecular basis of adaptation to intracellular macrophage environment.
Highlights
Tuberculosis (TB), caused by Mycobacterium tuberculosis is a devastating health problem
The results indicate that the present in vitro infection model was able to support the growth of M. bovis BCG in murine macrophages and is an ideal model to determine the pattern of functions of gene expression during the interaction of mycobacterium with macrophages
Our results with thymidine uptake assay clearly suggest that the present in vitro infection model was able to support the growth of M. bovis BCG in murine macrophages
Summary
Tuberculosis (TB), caused by Mycobacterium tuberculosis is a devastating health problem. World Health Organization (WHO) estimates that 2 billion people have latent TB while another 3 million people worldwide die of TB each year [1]. Resurgence of tuberculosis has intensified the necessity for new approaches to combat M. tuberculosis. M. tuberculosis is a facultative, intracellular pathogen which resides within the macrophages of the host. To survive intracellularly it must respond to the intracellular milieu of the macrophages. It has been well documented that intraphagosomal survival and growth of mycobacteria in macrophage is associated with changes in their gene expression and protein composition upon engulfment [2,3,4]
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