Functional genomics of innate host defense molecules in normal human monocytes in response to Aspergillus fumigatus.
Aspergillus fumigatus induces the release of innate immune-related molecules from phagocytic cells early in the course of infection. Little is known, however, about the complex expression profiles of the multiple genes involved in this response. We therefore investigated the kinetics of early gene expression in human monocytes (HMCs) infected with conidia of A. fumigatus using DNA microarray analysis. Total RNA from HMCs at 0, 2, 4, and 6 h was extracted, linearly amplified, hybridized onto Affymetrix HG133 Plus 2.0 gene chips, and analyzed with an Affymetrix scanner. Changes in gene expression were calculated as a ratio of those expressed by infected versus control HMCs. Aspergillus fumigatus induced differential regulation of expression in 1,827 genes (P < 0.05). Genes encoding cytokines and chemokines involved in host defense against A. fumigatus, including interleukin-1beta (IL-1beta), IL-8, CXCL2, CCL4, CCL3, and CCL20, as well as the opsonin long pentraxin 3, were up-regulated during the first 2 to 6 h, coinciding with an increase in phagocytosis. Simultaneously, genes encoding CD14, ficolin1, and MARCO were down-regulated, and genes encoding IL-10 and matrix metalloproteinase 1 were up-regulated. Up-regulation of the genes encoding heat shock proteins 40 and 110 and connexins 26 and 30 may point to novel molecules whose role in the pathogenesis of aspergillosis has not been previously reported. Verification of the transcriptional profiling was obtained for selected genes by reverse transcription-PCR and enzyme immunoassay. Thus, A. fumigatus conidia induced a coordinated expression of genes important in host defense and immunomodulation.
Cortez
Karoll J
KJ
Immunocompromised Host Section, Pediatric Oncology Branch, National Cancer Institute, NIH, 10 Center Drive, CRC Rm. 1-5250, Bethesda, MD 20892-1882, USA.
Lyman
Caron A
CA
Kottilil
Shyam
S
Kim
Hee Sup
HS
Roilides
Emmanuel
E
Yang
Jun
J
Fullmer
Brandie
B
Lempicki
Richard
R
Walsh
Thomas J
TJ
D001228:Aspergillosis / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D001232:Aspergillus fumigatus / Q000276:immunology*; D002470:Cell Survival / Q000235:genetics / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D020869:Gene Expression Profiling; D023281:Genomics / Q000379:methods; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics*; D007700:Kinetics; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis / Q000235:genetics; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011453:Prostaglandins / Q000096:biosynthesis / Q000235:genetics; D018121:Receptors, Cytokine / Q000096:biosynthesis / Q000235:genetics; D051192:Receptors, Pattern Recognition / Q000096:biosynthesis / Q000235:genetics
D016207:Cytokines; D011453:Prostaglandins; D018121:Receptors, Cytokine; D051192:Receptors, Pattern Recognition; D020781:Matrix Metalloproteinase 1