Changes in gene expression in macrophages infected with Mycobacterium tuberculosis: a combined transcriptomic and proteomic approach

Abstract

SUMMARYWe investigated the changes which occur in gene expression in the human macrophage cell line, THP1, at 1, 6 and 12 hr following infection with Mycobacterium tuberculosis. The analysis was carried out at the transcriptome level, using microarrays consisting of 375 human genes generally thought to be involved in immunoregulation, and at the proteomic level, using two-dimensional gel electrophoresis and mass spectrometry. The analysis of the transcriptome using microarrays revealed that many genes were up-regulated at 6 and 12 hr. Most of these genes encoded proteins involved in cell migration and homing, including the chemokines interleukin (IL)-8, osteopontin, monocyte chemotactic protein-1 (MCP-1), macrophage in¯ammatory protein-1a (MIP-1a), regulated on activation, normal, T-cell expressed and secreted (RANTES), MIP-1b, MIP-3a, myeloid progenitor inhibitory factor-1 (MPIF-1), pulmonary and activation regulated chemokine (PARC), growth regulated gene-b (GRO-b), GRO-c, MCP-2, I-309, and the T helper 2 (Th2) and eosinophil-attracting chemokine, eotaxin. Other genes involved in cell migration which were up-regulated included the matrix metalloproteinase MMP-9, vascular endothelial growth factor (VEGF) and its receptor Flk-1, the chemokine receptor CCR3, and the cell adhesion molecules vesicular cell adhesion molecule-1 (VCAM-1) and integrin a3. In addition to the chemokine response, genes encoding the proin¯ammatory cytokines IL-1b (showing a 433-fold induction), IL-2 and tumour necrosis factor-a (TNF-a), were also found to be induced at 6 and/or 12 hr. It was more dif®cult to detect changes using the proteomic approach. Nevertheless, IL-1b was again shown to be strongly up-regulated. The enzyme manganese superoxide dismutase was also found to be strongly up-regulated; this enzyme was found to be macrophage-, rather than M. tuberculosis, derived. The heat-shock protein hsp27 was found to be down-regulated following infection. We also identi®ed a mycobacterial protein, the product of the atpD gene (thought to be involved in the regulation of cytoplasmic pH) in the infected macrophage extracts.

Patterns of Chemokine Receptor Expression on Peripheral Blood γδ T Lymphocytes: Strong Expression of CCR5 Is a Selective Feature of Vδ2/Vγ9 γδ T Cells

“…8) might be due to the fact that IPP is recognized exclusively by V␦2V␥9 ␥␦ T cells within the PBMC, while the M. tb. lysate used in this study also activates monocytes/macrophages (36). Indeed, our further analysis with E-rosette-purified T cells confirmed this assumption, because CCR5 down-modulation on ␥␦ T cells in response to M. tb.…”

“…induced a much stronger CCR5 downmodulation than IPP on ␥␦ T cells when PBMC were used as responder cells suggested that the modulation triggered by M. tb. might be an indirect effect, in part due to the M. tb.-induced macrophage activation and subsequent cytokine and/or chemokine production by macrophages (36). To address this issue, we compared the CCR5 modulation on gated ␥␦ T cells when PBMC (containing ϳ30% monocytes) or E-rosette-purified T cells were used as responder cells.…”

The Chemokine Receptor CXCR3 Attenuates the Control of Chronic Mycobacterium tuberculosis Infection in BALB/c Mice

“…8E). ICAM-1, shown to be of importance in mediating granuloma formation and subsequent tubercle bacillary containment as a means of controlling their dissemination and replication (41), was expressed to similar levels in M. tuberculosisinfected CXCR3-deficient and wild-type BALB/c mice, as were other inflammatory mediators like matrix metalloproteinase 9 (MMP-9), tissue inhibitor of metalloproteinase 2 (TIMP-2), and secreted phosphoprotein (Spp) 1 (42)(43)(44)(45) (Fig. 8E).…”

Expression of immunoregulatory genes in peripheral blood mononuclear cells of European wild boar immunized with BCG

“…RANTES is a proinflammatory chemokine that plays an active role in recruiting leukocytes into inflammatory sites and 25 also induces the proliferation and activation of certain natural-killer (NK) cells to form A c c e p t e d M a n u s c r i p t Vet Microbiol 9 CHAK (CC-Chemokine-activated killer) cells (Maghazachi et al, 1996). RANTES is induced by M. tuberculosis in mouse and human macrophages (Ehrt et al, 2001;Ragno et al, 2001) and in mouse lungs after aerogenic infection (Keller et al, 2006). RANTES is also weakly induced in BCG vaccinated guinea pigs (Tree et al, 2006).…”