Gene expression profiling of monocyte-derived macrophages following infection withMycobacterium aviumsubspeciesaviumandMycobacterium aviumsubspeciesparatuberculosis

Abstract

Mycobacterium avium subspecies paratuberculosis (MAP) and Mycobacterium avium subspecies avium (MAA) represent two closely related intracellular bacteria with vastly different associated pathologies. MAA can cause severe respiratory infections in immune compromised humans but is nonpathogenic in ruminants and is more readily controlled by the bovine immune system than MAP. MAP causes a fatal wasting syndrome in ruminants, typified by granulomatous enteritis localized in the small intestine. MAP has also been cited as a potential cause of human Crohn's disease. We used a bovine immune-specific microarray (BOTL-5) to compare the response of mature bovine monocyte-derived macrophages (MDM cells) to MAP and MAA. Statistical analysis of microarray data revealed 21 genes not appreciably expressed in resting MDM cells that were activated following infection with either MAA or MAP. Further analysis revealed 144 genes differentially expressed in MDM cells following infection with MAA and 99 genes differentially expressed following infection with MAP. Of these genes, 37 were affected by both types of mycobacteria, with three being affected in opposite directions. Over 41% of the differentially expressed genes in MAA and MAP infected MDM cells were members of, regulated by, or regulators of the MAPK pathways. Expression of selected genes was validated by quantitative real-time reverse transcriptase PCR and in several key genes (i.e., IL-2 receptor, tissue inhibitor of matrix metalloproteinases-1, and Fas-ligand) MAA was found to be a stronger activating factor than MAP. These gene expression patterns were correlated with prolonged activation of p38 MAPK and ERK1/2 by MAA, relative to MAP.

Macrophage plasticity and polarization: in vivo veritas

“…M1 macrophages are generally considered responsible for resistance against intracellular pathogens and characterize infection with Listeria monocytogenes (55), Salmonella typhi, and Salmonella typhimurium (56), as well as the early phases of infection with Mycobacterium tuberculosis (57), Mycobacterium ulcerans, and Mycobacterium avium (58,59). Uncontrolled M1 inflammation associated with acute infections with E. coli or Streptococcus sp.…”

“…In a murine model of experimental colitis, recruitment of CCL11-expressing Ly6C hi CCR2 + inflammatory monocytes into the colon correlates with eosinophil infiltration and histopathology (54). Mycobacterium ulcerans, and Mycobacterium avium (58,59). Uncontrolled M1 inflammation associated with acute infections with E. coli or Streptococcus sp.…”

Macrophage Polarization in Bacterial Infections

“…However, a small subset of tuberculosis patients is characterized by M2-type patterns, which can be reversed by antibiotic treatment (30). Other mycobacterial diseases such as Buruli disease (Mycobacterium ulcerans) and opportunistic infections (Mycobacterium avium) are also characterized and controlled by M1 polarization of macrophages (31,32).…”

Oligonucleotide Microarray Technology and its Application to Mycobacterium avium subsp. paratuberculosis Research: A Review

“…However, in the case of clinical and food analysis, in which time is a critical factor, microarray technology can represent a suitable tool [50][51][52]. Another benefit of arrays is their Bovine cDNA microarray MAP infected cows cDNA from ileal tissue [27] Bovine cDNA microarray MAP infected bovine macrophages cDNA from macrophages [91,93] Human GeneChip (Affymetrix) MAP infected bovine macrophages cRNA (from cDNA) from macrophages [92] MAP whole genome array MAP infected bovine epithelial cells cDNA from epithelial cells [96] Bovine cDNA microarray Leucocytes from MAP infected cows cDNA from leucocytes [97] MAP whole genome array MAP (from cattle, sheep and human) infected human macrophages cRNA (from cDNA) from macrophages [102] Notes: cDNA complementary DNA, cRNA complementary RNA, gDNA genomic DNA, MAA Mycobacterium avium subsp. avium, MAP Mycobacterium avium subsp.…”