Gene expression profiles of peripheral blood leukocytes after endotoxin challenge in humans.

To define gene expression profiles that occur during the initial activation of human innate immunity, we administered intravenous endotoxin (n = 8) or saline (n = 4) to healthy subjects and hybridized RNA from blood mononuclear cells (0, 0.5, 6, 24, 168 h) or whole blood (0, 3, 6, 24, 168 h) to oligonucleotide probe arrays. The greatest change in mononuclear cell gene expression occurred at 6 h (439 induced and 428 repressed genes, 1% false discovery rate, and 50% fold change) including increased expression of genes associated with pathogen recognition molecules and signaling cascades linked to receptors associated with cell mobility and activation. Induced defense response genes included cytokines, chemokines, and their respective receptors, acute-phase transcription factors, proteases, arachidonate metabolites, and oxidases. Repressed defense response genes included those associated with co-stimulatory molecules, T and cytotoxic lymphocytes, natural killer (NK) cells, and protein synthesis. Gene expression profiles of whole blood had similar biological themes. Over 100 genes not typically associated with acute inflammation were differentially regulated after endotoxin. By 24 h, gene expression had returned to baseline values. Thus the inflammatory response of circulating leukocytes to endotoxin in humans is characterized by a rapid amplification and subsidence of gene expression. These results indicate that a single intravascular exposure to endotoxin produces a large but temporally short perturbation of the blood transcriptome.

[1]  J. Kovacs,et al.  The cardiovascular response of normal humans to the administration of endotoxin. , 1989, The New England journal of medicine.

[2]  Mark Hurle,et al.  Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects. , 2005, DNA and cell biology.

[3]  H. Baker,et al.  Molecular Characterization of the Acute Inflammatory Response to Infections with Gram-Negative versus Gram-Positive Bacteria , 2003, Infection and Immunity.

[4]  B. Beutler.  Innate immunity: an overview. , 2004, Molecular immunology.

[5]  Jason Laramie,et al.  Whole blood and leukocyte RNA isolation for gene expression analyses. , 2004, Physiological genomics.

[6]  B Alex Merrick,et al.  Differential gene expression profiling in whole blood during acute systemic inflammation in lipopolysaccharide-treated rats. , 2005, Physiological genomics.

[7]  A C Allison,et al.  The acute splanchnic and peripheral tissue metabolic response to endotoxin in humans. , 1990, The Journal of clinical investigation.

[8]  J. Cavaillon,et al.  Toll-like receptor-mediated tumor necrosis factor and interleukin-10 production differ during systemic inflammation. , 2003, American journal of respiratory and critical care medicine.

[9]  L. Staudt,et al.  Gene expression physiology and pathophysiology of the immune system. , 2001, Trends in immunology.

[10]  J. Pugin,et al.  Normal responses to injury prevent systemic inflammation and can be immunosuppressive. , 2001, American journal of respiratory and critical care medicine.

[11]  Mehmet Toner,et al.  Application of genome-wide expression analysis to human health and disease. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[12]  P. Brown,et al.  Genomic views of the immune system*. , 2000, Annual review of immunology.

[13]  T. van der Poll,et al.  Reduced Th1, but Not Th2, Cytokine Production by Lymphocytes after In Vivo Exposure of Healthy Subjects to Endotoxin , 2000, Infection and Immunity.

[14]  A. Coutinho,et al.  Administration to mouse of endotoxin from gram‐negative bacteria leads to activation and apoptosis of T lymphocytes , 1998, European journal of immunology.

[15]  M. Hellmich,et al.  Comparison of different isolation techniques prior gene expression profiling of blood derived cells: impact on physiological responses, on overall expression and the role of different cell types , 2004, The Pharmacogenomics Journal.

[16]  John I. Gallin,et al.  Inflammation: Basic Principles and Clinical Correlates , 1992 .

[17]  H. Brade,et al.  Endotoxin in Health and Disease , 2020 .

[18]  P. Essén,et al.  Response of in vivo protein synthesis in T lymphocytes and leucocytes to an endotoxin challenge in healthy volunteers , 2002, Clinical and experimental immunology.

[19]  M J Tobin,et al.  Effect of endotoxin on ventilation and breath variability: role of cyclooxygenase pathway. , 2001, American journal of respiratory and critical care medicine.

[20]  J. Cavaillon,et al.  Endotoxin tolerance: is there a clinical relevance? , 2003, Journal of endotoxin research.

[21]  Charles Natanson,et al.  Meta-Analysis: The Effect of Steroids on Survival and Shock during Sepsis Depends on the Dose , 2004, Annals of Internal Medicine.

[22]  Djillali Annane,et al.  Septic shock , 2005, The Lancet.

[23]  W. Knaus,et al.  Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. The ACCP/SCCM Consensus Conference Committee. American College of Chest Physicians/Society of Critical Care Medicine. , 1992, Chest.

[24]  Douglas M. Hawkins,et al.  A variance-stabilizing transformation for gene-expression microarray data , 2002, ISMB.

[25]  J. White,et al.  Effects of anti-inflammatory agents on serum levels of calcitonin precursors during human experimental endotoxemia. , 2001, The Journal of infectious diseases.

[26]  D. Alling,et al.  COMPARISON OF HEMATOLOGIC AND FEBRILE RESPONSE TO ENDOTOXIN IN MAN. , 1965, Blood.

[27]  Ash A. Alizadeh,et al.  Stereotyped and specific gene expression programs in human innate immune responses to bacteria , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  S. Pillai,et al.  Innate immunity. , 1996, Current opinion in immunology.

[29]  David E. Misek,et al.  Profiling Changes in Gene Expression during Differentiation and Maturation of Monocyte-derived Dendritic Cells Using Both Oligonucleotide Microarrays and Proteomics* , 2001, The Journal of Biological Chemistry.

[30]  E. Lander,et al.  Human macrophage activation programs induced by bacterial pathogens , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[31]  F. Schildberg,et al.  Downregulation of proinflammatory cytokine release in whole blood from septic patients. , 1995, Blood.

[32]  John D. Storey,et al.  A network-based analysis of systemic inflammation in humans , 2005, Nature.

[33]  P. Hegde,et al.  The transcriptome in blood: challenges and solutions for robust expression profiling. , 2005, Current molecular medicine.

[34]  D. Mannino,et al.  The epidemiology of sepsis in the United States from 1979 through 2000. , 2003, The New England journal of medicine.

[35]  J. Siewert,et al.  Distinct mechanisms of immunosuppression as a consequence of major surgery , 1997, Infection and immunity.

[36]  R. Hotchkiss,et al.  The pathophysiology and treatment of sepsis. , 2003, The New England journal of medicine.

[37]  Charles Natanson,et al.  Risk and the efficacy of antiinflammatory agents: retrospective and confirmatory studies of sepsis. , 2002, American journal of respiratory and critical care medicine.

[38]  J. Helterbrand,et al.  Efficacy and safety of recombinant human activated protein C for severe sepsis , 2003 .

[39]  T. van der Poll,et al.  Modulation of cytokine release and neutrophil function by granulocyte colony-stimulating factor during endotoxemia in humans. , 1997, Blood.

[40]  K. Asadullah,et al.  Monocyte deactivation in septic patients: restoration by IFN-gamma treatment. , 1997, Nature medicine.

[41]  H. D. Hochstein,et al.  Dose-related inflammatory effects of intravenous endotoxin in humans: evaluation of a new clinical lot of Escherichia coli O:113 endotoxin. , 1999, The Journal of infectious diseases.

[42]  P. Marrack,et al.  Activation changes the spectrum but not the diversity of genes expressed by T cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[43]  M. Gladwin,et al.  Blood mononuclear cell gene expression profiles characterize the oxidant, hemolytic, and inflammatory stress of sickle cell disease. , 2004, Blood.

[44]  Carl Nathan,et al.  Points of control in inflammation , 2002, Nature.

[45]  A. Dietz,et al.  Maturation of human monocyte-derived dendritic cells studied by microarray hybridization. , 2000, Biochemical and biophysical research communications.

[46]  M. Meisner,et al.  Induction of Procalcitonin and Proinflammatory Cytokines in an Anhepatic Baboon Endotoxin Shock Model , 2003, Shock.

[47]  A. Eerenberg,et al.  Inhibition of endotoxin-induced cytokine release and neutrophil activation in humans by use of recombinant bactericidal/permeability-increasing protein. , 1995, The Journal of infectious diseases.

[48]  B. Beutler,et al.  Innate immune sensing and its roots: the story of endotoxin , 2003, Nature Reviews Immunology.

[49]  Joshua M. Korn,et al.  The plasticity of dendritic cell responses to pathogens and their components. , 2001, Science.

[50]  Yoav Benjamini,et al.  Identifying differentially expressed genes using false discovery rate controlling procedures , 2003, Bioinform..

[51]  A. von Ruecker,et al.  Surgical stress induces a shift in the type-1/type-2 T-helper cell balance, suggesting down-regulation of cell-mediated and up-regulation of antibody-mediated immunity commensurate to the trauma. , 1996, Surgery.