Gene Expression Profiles Of Macrophages Research Articles

Regulation of gene expression in mouse macrophages stimulated with bacterial CpG-DNA and lipopolysaccharide

CpG-DNA is known as a potent immunostimulating agent and may contribute in therapeutic treatment of many immune disorders. CpG-DNA triggers innate and acquired immune responses through activated expression of various genes in immune cells, including macrophages. To define the molecular mechanism(s) by which CpG-DNA activates immune cells, we studied macrophage gene expression following CpG-DNA exposure using high-density oligonucleotide microarrays. As CpG-DNA receptor Toll-like receptor 9 (TLR9) shares homology with the lipopolysaccharide (LPS)-TLR4 receptor, we compared gene expression profiles in macrophages stimulated by LPS versus CpG-DNA. CpG-DNA and LPS modulate expression of many genes encoding cytokines, cell surface receptors, transcription factors, and proteins related to cell proliferation/differentiation. However, LPS modulated expression of significantly more genes than did CpG-DNA, and all genes induced or repressed by CpG-DNA were induced or repressed by LPS. We conclude that CpG-DNA signaling through TLR9 activates a subset of genes induced by LPS-TLR4 signaling.

Survival tactics of M. paratuberculosis in bovine macrophage cells

Mycobacterium paratuberculosis ( M. paratuberculosis) is a facultative intracellular bacterium with the ability to survive and proliferate inside the vesicles of macrophage cells. How M. paratuberculosis and other mycobacteria survive in this hostile environment is not well understood. Present research findings can be divided into three possibilities: (1) mycobacteria may interfere with host protein expression and trafficking to stop vesicle maturation, (2) mycobacteria may express proteins that interfere with macrophage activation in a more direct manner, or (3) mycobacteria may enter macrophages in such a way as to avoid the normal process of activation via Toll-like receptors and other, as yet unknown mechanisms. Research thus far has predominately centered on possible macrophage/mycobacteria protein interactions. To more completely define how mycobacteria interfere with the process of phagosome maturation our group has recently taken a functional genomics approach, allowing the macrophage to “tell” us what host genes may be affected by phagocytosis of mycobacteria. We used DDRT-PCR to examine differences in macrophage cell gene expression during phagocytosis of E. coli and M. paratuberculosis. Macrophage cells not exposed to any phagocytosis target and in the process of phagocytosing latex beads were used as negative and positive controls, respectively. To date, we have identified 380 DDRT-PCR amplicons corresponding to transcripts whose expression profiles appear to be altered during the general process of phagocytosis. Dot-blot and Northern blot hybridizations with a subset of these amplicons were performed to confirm results observed with DDRT-PCR. Our preliminary results indicate that macrophage gene expression profiles change dramatically following phagocytosis and that gene expression profiles following phagocytosis of M. paratuberculosis are different than those following phagocytosis of E. coli or latex beads.

Salmonella typhimurium infection and lipopolysaccharide stimulation induce similar changes in macrophage gene expression.

Changes in macrophage phenotype induced during infection result from the recognition of bacterial products as well as the action of bacterial virulence factors. We used the unprecedented opportunity provided by gene arrays to simultaneously study the expression of hundreds of genes during Salmonella typhimurium infection of macrophages and to assess the contribution of the bacterial virulence factor, LPS, in initiating the host responses to Salmonella. We found that S. typhimurium infection caused significant changes in the expression of numerous genes encoding chemokines, cell surface receptors, signaling molecules, and transcriptional activators at 4 h postinfection of the RAW 264.7 murine macrophage cell line. Our results revealed changes in the expression of several genes that had not been previously implicated in the host responses to S. typhimurium infection, as well as changes in the expression of several genes previously shown to be regulated by S. typhimurium infection. An overlapping spectrum of genes was expressed in response to virulent S. typhimurium and purified S. typhimurium LPS, reinforcing the major role of this surface molecule in stimulating the early response of macrophages to bacterial infection. The macrophage gene expression profile was further altered by activation with IFN-gamma, indicating that host cell responses depend on the activation state of the cell.

Determinants of high density lipoprotein and total cholesterol in women.

An epidemiological study was carried out among a random sample of women aged 18 to 69 years to examine possible determinants of plasma high density lipoprotein and total cholesterol (HDL-C and T-C). In a multiple regression analysis consumption of alcohol, fatty fish, and parental longevity showed positive associations with HDL-C, which were statistically significant. Smoking habit, sucrose consumption, and a family history of ischaemic heart disease showed significantly negative associations. In contrast, T-C was associated significantly only with age and Quetelet's index of body mass. Five variables having significant associations with HDL-C explained only 8% of the total variance; in the case of T-C 17% of the variance was explained by age and body mass.