An in vitro infection model system to study proteins expressed during interaction of mycobacterium with murine macrophages
- 1
- 2
- 3
- 4
- 5
- 6
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.
- World Health Organization (2008) Global tuberculosis control: Surveillance, planning, financing. WHO Report 2008. http://www.who.int/tb/publications/global_report/ 2008/en/index.html
- McDonough, K.A., Florczyk, M.A. and Kress, Y. (2000) Intracellular passage within macrophages affects the trafficking of virulent tubercle bacilli upon reinfection of other macrophages in a serum-dependent manner. Tubercle and Lung Disease, 80(6), 259-271.
- Monahan, I.M., Betts, J., Banerjee, D.K. and Butcher, P.D. (2001) Differential expression of mycobacterial proteins following phagocytosis by macrophages. Microbiology, 147(2), 459-471.
- Schnappinger, D., Ehrt, S., Voskuil, M.I., Liu, Y., Mangan, J.A., Monahan, I.M., Dolganov, G., Efron, B., Butcher, P.D., Nathan C. and Schoolnik, G.K. (2003) Transcriptional adaptation of Mycobacterium tuberculosis within macrophages: Insights into the phagosomal environment. Journal of Experimental Medicine, 198(5), 693-704.
- Jungblut, P.R., Schaible, U.E., Mollenkopf, H.J., Zimny- Arndt, U., Raupach, B., Mattow, J., Halada, P., Lamer, S., Hagens K. and Kaufmann, S.H. (1999) Comparative proteome analysis of Mycobacterium tuberculosis and Mycobacterium bovis BCG strains: Towards functional genomics of microbial pathogens. Molecular Microbiology, 33(6), 1103-1117.
- Rosenkrands, I., Weldingh, K., Jacobsen, S., Hansen, C.V., Florio, W., Gianetri, I. and Andersen, P. (2000) Mapping and identification of Mycobacterium tuberculosis proteins by two-dimensional gel electrophoresis, mi- crosequencing and immunodetection. Electrophoresis, 21(5), 935-948.
- Sinha, S., Arora, S., Kosalai, K., Namane, A., Pym, A.S. and Cole, S.T. (2005) Proteome analysis of the plasma membrane of mycobacterium tuberculosis. Microbiology, 151(7), 2411-2419.
- Mattow, J., Siejak, F., Hagens, K., Becher, D., Albrecht, D., Krah, A., Schmidt, F., Jungblut, P.R., Kaufmann, S.H. and Schaible, U.E. (2006) Proteins unique to intraphago- somally grown Mycobacterium tuberculosis. Proteomics, 6(8), 2485-2494.
- Barker, K., Fan, H., Carroll, C., Kaplan, G., Barker, J., Hellmann, W. and Cohn, Z.A. (1996) Nonadherent cultures of human monocytes kill Mycobacterium smegmatis, but adherent cultures do not. Infection Immunity, 64(2), 428-433.
- Mehta, P.K., King, C.H., White, E.H., Murtagh, J., Jr. and Quinn, F.D. (1996) Comparison of in vitro models for the study of Mycobacterium tuberculosis invasion and intracellular replication. Infection Immunity, 64(2), 2673- 2679.