Distinct gene expression profiles of human type 1 and type 2 T helper cells

BackgroundThe development and activation of CD4+ helper T cell (Th) subsets with distinct patterns of unbalanced production of cytokines play an important part in infectious, allergic and autoimmune diseases. Human neonatal cord blood CD4+ Th cells can be polarized into type 1 or type 2-like effector cells in vitro by culturing them in the presence of interleukin (IL)-12 or IL-4, respectively. We have exploited this experimental system to identify marker genes that are differentially expressed by polarized Th1 and Th2 cells. An oligonucleotide microarray specifically designed to screen for inflammation-related candidate genes was used and the differential expression was further validated with a quantitative real-time RT-PCR method.ResultsIn addition to the previously described marker genes of Th cells, we report subtle changes in the expression of several other genes that represent growth factors, receptors and other signaling molecules in polarized Th1 and Th2 cell subsets. Additionally, we describe a novel set of genes as Th1/Th2 differentiation markers for cells activated by anti-CD3 and anti-CD28 antibodies.ConclusionsThis study demonstrates the power of the targeted use of microarrays in combination with quantitative real-time RT-PCR in identifying and validating new marker genes for gene expression studies.

[1]  G. Peltz,et al.  Modulation of the Grb2-associated protein complex in human CD4+ T cells by receptor activation. , 1995, Journal of immunology.

[2]  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.

[3]  K. Matsushima,et al.  Selective recruitment of CCR4-bearing Th2 cells toward antigen-presenting cells by the CC chemokines thymus and activation-regulated chemokine and macrophage-derived chemokine. , 1999, International immunology.

[4]  B. Williams,et al.  Identification of genes differentially regulated by interferon α, β, or γ using oligonucleotide arrays , 1998 .

[5]  D. Green,et al.  Unequal Death in T Helper Cell (Th)1 and Th2 Effectors: Th1, but not Th2, Effectors Undergo Rapid Fas/FasL-mediated Apoptosis , 1997, The Journal of experimental medicine.

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

[7]  J. Carver,et al.  Clusterin Has Chaperone-like Activity Similar to That of Small Heat Shock Proteins* , 1999, The Journal of Biological Chemistry.

[8]  Kenneth M. Murphy,et al.  Functional diversity of helper T lymphocytes , 1996, Nature.

[9]  M. Holtzman,et al.  Resistance to Fas-mediated T cell apoptosis in asthma. , 1999, Journal of immunology.

[10]  Jan Mous,et al.  Touching base , 2000, Nature Genetics.

[11]  J. Hodgson,et al.  DNA chips: An array of possibilities , 1998, Nature Biotechnology.

[12]  Ash A. Alizadeh,et al.  Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling , 2000, Nature.

[13]  C. Heid,et al.  A novel method for real time quantitative RT-PCR. , 1996, Genome research.

[14]  R. Locksley,et al.  Coordinate Regulation of the IL-4, IL-13, and IL-5 Cytokine Cluster in Th2 Clones Revealed by Allelic Expression Patterns1 , 2000, The Journal of Immunology.

[15]  M. Biffi,et al.  Selective Expression of an Interleukin-12 Receptor Component by Human T Helper 1 Cells , 1997, Journal of Experimental Medicine.

[16]  C. Mackay,et al.  Flexible Programs of Chemokine Receptor Expression on Human Polarized T Helper 1 and 2 Lymphocytes , 1998, The Journal of experimental medicine.

[17]  G. Delespesse,et al.  Default development of cloned human naive CD4 T cells into interleukin‐4‐ and interleukin‐5‐producing effector cells , 1995, European journal of immunology.

[18]  B. Williams,et al.  Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Biffi,et al.  Selective Expression of an Interleukin-12 Receptor Component by Human T Helper 1 Cells , 1997, The Journal of experimental medicine.

[20]  D. Lockhart,et al.  Expression monitoring by hybridization to high-density oligonucleotide arrays , 1996, Nature Biotechnology.

[21]  M. Tomlinson,et al.  TGF‐β1 down‐regulates Th2 development and results in decreased IL‐4‐induced STAT6 activation and GATA‐3 expression , 2000 .

[22]  G. Peltz,et al.  CD45 isoforms on human CD4+ T-cell subsets. , 1996, The Journal of allergy and clinical immunology.

[23]  Richard A Flavell,et al.  The Transcription Factor GATA-3 Is Necessary and Sufficient for Th2 Cytokine Gene Expression in CD4 T Cells , 1997, Cell.

[24]  A. Hamann,et al.  T helper 1 and T helper 2 cells respond differentially to chemokines. , 1998, Journal of immunology.

[25]  R. Flavell,et al.  Cutting Edge: TGF-β Inhibits Th Type 2 Development Through Inhibition of GATA-3 Expression , 2000, The Journal of Immunology.

[26]  Suneet Agarwal,et al.  Modulation of chromatin structure regulates cytokine gene expression during T cell differentiation. , 1998, Immunity.

[27]  B Dewald,et al.  Human chemokines: an update. , 1997, Annual review of immunology.

[28]  R. Bucala,et al.  An essential regulatory role for macrophage migration inhibitory factor in T-cell activation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. Bittner,et al.  Expression profiling using cDNA microarrays , 1999, Nature Genetics.

[30]  J. Mesirov,et al.  Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. , 1999, Science.

[31]  David G. Morris,et al.  Global analysis of gene expression in pulmonary fibrosis reveals distinct programs regulating lung inflammation and fibrosis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[32]  R. W. Davis,et al.  Discovery and analysis of inflammatory disease-related genes using cDNA microarrays. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[33]  S. P. Fodor,et al.  High density synthetic oligonucleotide arrays , 1999, Nature Genetics.

[34]  R. Coffman,et al.  Induction of human T helper cell type 1 differentiation results in loss of IFN-gamma receptor beta-chain expression. , 1997, Journal of immunology.

[35]  R. Lahesmaa,et al.  Signaling lymphocytic activation molecule (SLAM) is differentially expressed in human Th1 and Th2 cells. , 2000, Journal of immunological methods.

[36]  N. Kadowaki,et al.  The nature of the principal type 1 interferon-producing cells in human blood. , 1999, Science.

[37]  Laurie H Glimcher,et al.  The Proto-Oncogene c-maf Is Responsible for Tissue-Specific Expression of Interleukin-4 , 1996, Cell.

[38]  S. Swain,et al.  Reciprocal regulation of polarized cytokine production by effector B and T cells , 2000, Nature Immunology.

[39]  P. Zipfel,et al.  Synthesis of the CC-chemokines MIP-1alpha, MIP-1beta, and RANTES is associated with a type 1 immune response. , 1996, Journal of immunology.

[40]  G. Miller,et al.  Costimulation of human CD4+ T cells by fibroblast growth factor-1 (acidic fibroblast growth factor). , 1995, Journal of immunology.

[41]  L. Glimcher,et al.  Lineage commitment in the immune system: the T helper lymphocyte grows up. , 2000, Genes & development.

[42]  P. Salgame,et al.  Differential ability of T cell subsets to undergo activation-induced cell death. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[43]  Roger E Bumgarner,et al.  Large-scale monitoring of host cell gene expression during HIV-1 infection using cDNA microarrays. , 2000, Virology.

[44]  P. Allavena,et al.  Differential Expression of Chemokine Receptors and Chemotactic Responsiveness of Type 1 T Helper Cells (Th1s) and Th2s , 1998, The Journal of experimental medicine.

[45]  M. Salmon,et al.  IL-7-dependent extrathymic expansion of CD45RA+ T cells enables preservation of a naive repertoire. , 1998, Journal of immunology.

[46]  G. Berry,et al.  CD4(+) T helper cells engineered to produce latent TGF-beta1 reverse allergen-induced airway hyperreactivity and inflammation. , 2000, The Journal of clinical investigation.

[47]  T. Gingeras,et al.  Cellular gene expression altered by human cytomegalovirus: global monitoring with oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[48]  S. Dzik.  The nature of the principal type 1 interferon-producing cells in human blood , 2000 .

[49]  R. Seder.  Acquisition of lymphokine-producing phenotype by CD4+ T cells. , 1994, The Journal of allergy and clinical immunology.

[50]  S. Romagnani.  Lymphokine production by human T cells in disease states. , 1994, Annual review of immunology.

[51]  M. Bittner,et al.  Fluorescent cDNA microarray hybridization reveals complexity and heterogeneity of cellular genotoxic stress responses , 1999, Oncogene.

[52]  M. Wilson,et al.  Effects of clusterin overexpression on TNFalpha- and TGFbeta-mediated death of L929 cells. , 1997, Biochemistry.

[53]  Laurie H Glimcher,et al.  A Novel Transcription Factor, T-bet, Directs Th1 Lineage Commitment , 2000, Cell.

[54]  A. Weinberg,et al.  Transforming growth factor-beta and IL-4 cause helper T cell precursors to develop into distinct effector helper cells that differ in lymphokine secretion pattern and cell surface phenotype. , 1991, Journal of immunology.

[55]  M. Baggiolini,et al.  CCR5 is characteristic of Th1 lymphocytes , 1998, Nature.

[56]  P. Marrack,et al.  Type I Interferons Keep Activated T Cells Alive , 1999, The Journal of experimental medicine.

[57]  Thomas D. Schmittgen,et al.  Real-Time Quantitative PCR , 2002 .