Monitoring Cellular Responses to Listeria monocytogenes with Oligonucleotide Arrays*

Listeria monocytogenes is a pathogenic intracellular microorganism whose infection induces pleiotropic biological changes associated with host cell gene expression regulation. Here we define the gene expression profiles of the human promyelocytic THP1 cell line before and after L. monocytogenes infection. Gene expression was measured on a large scale via oligonucleotide microarrays with probe sets corresponding to 6,800 human genes. We assessed and discussed the reproducibility of the hybridization signatures. In addition to oligonucleotide arrays, we also performed the large scale gene expression measurement with two high-density membranes, assaying for 588 and 18,376 human genes, respectively. This work allowed the reproducible identification of 74 up-regulated RNAs and 23 down-regulated RNAs as a consequence ofL. monocytogenes infection of THP1. The reliability of these data was reinforced by performing independent infections. Some of these detected RNAs were consistent with previous results, while some newly identified RNAs encode gene products that may play key roles inL. monocytogenes infection. These findings will undoubtedly enhance the understanding of L. monocytogenes molecular physiology and may help identify new therapeutic targets.

[1]  L. Wodicka,et al.  Genome-wide expression monitoring in Saccharomyces cerevisiae , 1997, Nature Biotechnology.

[2]  B. Jordan.  Large-scale expression measurement by hybridization methods: from high-density membranes to "DNA chips". , 1998, Journal of biochemistry.

[3]  A. Aruffo,et al.  4-1BB Costimulatory Signals Preferentially Induce CD8+ T Cell Proliferation and Lead to the Amplification In Vivo of Cytotoxic T Cell Responses , 1997, The Journal of experimental medicine.

[4]  P. Brown,et al.  Drug target validation and identification of secondary drug target effects using DNA microarrays , 1998, Nature Medicine.

[5]  W. Goebel,et al.  Differential regulation of cytokine and cytokine receptor mRNA expression upon infection of bone marrow-derived macrophages with Listeria monocytogenes , 1996, Infection and immunity.

[6]  G. Lennon,et al.  Hybridization analyses of arrayed cDNA libraries. , 1991, Trends in genetics : TIG.

[7]  J. Kappes,et al.  The Cysteine-Cysteine Family of Chemokines RANTES, MIP-1α, and MIP-1β Induce Trypanocidal Activity in Human Macrophages via Nitric Oxide , 1998, Infection and Immunity.

[8]  D. Botstein,et al.  The transcriptional program in the response of human fibroblasts to serum. , 1999, Science.

[9]  A. Pardee,et al.  Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. , 1992, Science.

[10]  D. Drevets,et al.  Listeria monocytogenes infection and activation of human brain microvascular endothelial cells. , 1998, The Journal of infectious diseases.

[11]  S. Klebanoff.  The iron-H2O2-iodide cytotoxic system , 1982, The Journal of experimental medicine.

[12]  P. Casellas,et al.  Stimulation of Cannabinoid Receptor CB1 Induces krox-24 Expression in Human Astrocytoma Cells (*) , 1995, The Journal of Biological Chemistry.

[13]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[14]  P. Cossart,et al.  Interactions of Listeria monocytogenes with mammalian cells during entry and actin‐based movement: bacterial factors, cellular ligands and signaling , 1998, The EMBO journal.

[15]  P. Casellas,et al.  Standardization of mRNA titration using a polymerase chain reaction method involving co-amplification with a multispecific internal control. , 1992, The Journal of biological chemistry.

[16]  Y Sakaki,et al.  High-density cDNA filter analysis: a novel approach for large-scale, quantitative analysis of gene expression. , 1995, Gene.

[17]  M. Soares,et al.  Construction and characterization of a normalized cDNA library. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[18]  S. Granjeaud,et al.  Differential gene expression in the murine thymus assayed by quantitative hybridization of arrayed cDNA clones. , 1995, Genomics.

[19]  J. Lancaster,et al.  EPR demonstration of iron-nitrosyl complex formation by cytotoxic activated macrophages. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[20]  L. Penland,et al.  Use of a cDNA microarray to analyse gene expression patterns in human cancer , 1996, Nature Genetics.

[21]  S H Kim,et al.  Exploiting chemical libraries, structure, and genomics in the search for kinase inhibitors. , 1998, Science.

[22]  P. Brown,et al.  Exploring the metabolic and genetic control of gene expression on a genomic scale. , 1997, Science.

[23]  R. Quatrano.  Genomics , 1998, Plant Cell.

[24]  C. Auffray,et al.  Novel gene transcripts preferentially expressed in human muscles revealed by quantitative hybridization of a high density cDNA array. , 1996, Genome research.

[25]  F. Matsumura,et al.  Fascin, an actin-bundling protein, induces membrane protrusions and increases cell motility of epithelial cells. , 1998, Molecular biology of the cell.

[26]  B. Finlay,et al.  Exploitation of mammalian host cell functions by bacterial pathogens. , 1997, Science.

[27]  G. Church,et al.  Genomic sequencing. , 1993, Methods in molecular biology.

[28]  Y. Sakaki,et al.  High-density cDNA filter analysis of the expression profiles of the genes preferentially expressed in human brain. , 1995, Gene.

[29]  T. Flotte,et al.  Expression of murine CD1 on gastrointestinal epithelium. , 1990, Science.