Sequential patterns of gene expression by bovine monocyte-derived macrophages associated with ingestion of mycobacterial organisms

We investigated mechanisms involved in killing of mycobacterial organisms by comparing the response of bovine monocyte-derived macrophages to ingestion of Mycobacterium avium subsp. paratuberculosis or M. avium subsp. avium organisms. Previous studies have shown that bovine macrophages have the capacity to kill M. avium subsp. avium organisms in vitro but cannot kill M. avium subsp. paratuberculosis organisms. We used bovine cDNA microarray technology to investigate sequential gene expression by bovine monocyte-derived macrophages and function assays to correlate gene expression with biological activity. Results of the gene expression studies indicated substantial differences between macrophages phagocytizing the two organisms. At 2, 6, and 24 h after infection, 12, 53, and 19 genes, respectively, were differentially expressed. Over all time periods, approximately twice as many genes had lower expression in M. avium subsp. paratuberculosis-infected macrophages than had greater expression. Differentially regulated genes of most interest to antimicrobial responses included inflammatory molecules (transforming growth factor-beta, thrombospondin 1, monocyte chemokine, and cathepsin K), phagosome-lysosome-related genes (H+ ATPases, lysosomal-associated membrane protein 2, vesicle trafficking protein, and solute carrier protein), and apoptosis-related genes (tumor necrosis factor receptor-associated factor 2, and tumor protein p 53 binding protein). Function assays indicated that M. avium subsp. avium-infected macrophages had a greater capacity to acidify phagosomes and a greater percentage of apoptotic cells. In conclusion, these results suggest that a complex interaction between macrophages and mycobacterial organisms is involved in determining the fate of the organism. Although multiple genes and metabolic pathways are involved, the capacity of cells to acidify phagosomes and induce apoptosis appears to play a prominent role. (C) 2004 Elsevier Ltd. All rights reserved.