Letter | Published:

Transcript imaging of the development of human T helper cells using oligonucleotide arrays

Nature Geneticsvolume 25pages96101 (2000) | Download Citation

Subjects

Abstract

Many pathological processes, including those causing allergies and autoimmune diseases, are associated with the presence of specialized subsets of T helper cells at the site of inflammation1. Understanding the genetic program that controls the functional properties of T helper type 1 (Th1) versus T helper type 2 (Th2) cells may provide insight into the pathophysiology of inflammatory diseases. We compared the gene-expression profiles of human Th1 and Th2 cells using high-density oligonucleotide arrays with the capacity to display transcript levels of 6,000 human genes2. Here we analyse the data sets derived from five independent experiments using statistical algorithms. This approach resulted in the identification of 215 differentially expressed genes, encoding proteins involved in transcriptional regulation, apoptosis, proteolysis, and cell adhesion and migration. A subset of these genes was further upregulated by exposure of differentiated Th1 cells to interleukin-12 (IL-12), as confirmed by kinetic PCR analysis, indicating that IL-12 modulates the effector functions of Th1 cells in the absence of antigenic stimulation. Functional assays and in vivo expression of selected genes have validated the biological relevance of our study. Our results provide new insight into the transcriptional program controlling the functional diversity of subsets of T helper cells.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Romagnani, S. Lymphokine production by human T cells in disease states. Annu. Rev. Immunol. 12, 227–257 (1994).

  2. 2

    Lipshutz, R.J., Fodor, S.P.A., Gingeras, T.R. & Lockhart, D.J. High density synthetic oligonucleotide arrays. Nature Genet. 21, 20–24 (1999).

  3. 3

    Abbas, A.K., Murphy, K.M. & Sher, A. Functional diversity of helper T lymphocytes. Nature 383, 787–793 (1996).

  4. 4

    O'Garra, A. Cytokines induce the development of functionally heterogenous T helper subsets. Immunity 8, 275–283 (1998).

  5. 5

    Gately, M.K. et al. The interleukin-12/interleukin-12-receptor system: role in normal and pathologic immune responses. Annu. Rev. Immunol. 16, 495–521 (1998).

  6. 6

    Seder, R.A. & Paul, W.E. Acquisition of lymphokine-producing phenotype by CD4+ T cells. Annu. Rev. Immunol. 12, 635–673 (1994).

  7. 7

    Trinchieri, G. Interleukin-12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Annu. Rev. Immunol. 13, 251–276 (1995).

  8. 8

    Rogge, L. et al. Selective expression of an interleukin-12 receptor component by human T helper 1 cells. J. Exp. Med. 185, 825–832 (1997).

  9. 9

    Rogge, L. et al. The role of Stat4 in species-specific regulation of T helper cell development by type I interferons. J. Immunol. 161, 6567–6574 (1998).

  10. 10

    Higuchi, R. & Watson, R. Kinetic PCR analysis using a CCD camera and without using oligonucleotide probes. in PCR Applications (eds Innis, M.A., Gelfand, D.H. & Sninsky, J.J.) 263–284 (Academic, San Diego, 1999).

  11. 11

    Zheng, W.-P. & Flavell, R.A. The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells. Cell 89, 587–596 (1997).

  12. 12

    Xu, D. et al. Selective expression and functions of interleukin 18 receptor on T helper (Th) type 1 but not Th2 cells. J. Exp. Med. 188, 1485–1492 (1998).

  13. 13

    Taki, S. et al. Multistage regulation of Th1-type immune responses by the transcription factor IRF-1. Immunity 6, 673–679 (1997).

  14. 14

    Lohoff, M. et al. Interferon regulatory factor-1 is required for a T helper 1 immune response in vivo. Immunity 6, 681–689 (1997).

  15. 15

    Coccia, E.M. et al. IL-12 induces expression of interferon regulatory factor-1 via signal transducer and activator of transcription-4 in human T helper type 1 cells. J. Biol. Chem. 274, 6698–6703 (1999).

  16. 16

    Austrup, F. et al. P- and E-selectin mediate recruitment of T-helper-1 but not T-helper-2 cells into inflamed tissues. Nature 385, 81–83 (1997).

  17. 17

    Butcher, E.C. & Picker, L.J. Leukocyte homing and homeostasis. Science 272, 60–66 (1996).

  18. 18

    Maly, P. et al. The α(1,3) fucosyltransferase Fuc-TVII controls leukocyte trafficking through an essential role in L-, E-, and P-selectin ligand biosynthesis. Cell 86, 643–653 (1996).

  19. 19

    Sallusto, F., Lanzavecchia, A. & Mackay, C.R. Chemokines and chemokine receptors in T-cell priming and Th1/Th2-mediated responses. Immunol. Today 19, 568–574 (1998).

  20. 20

    Van Damme, J. et al. The role of CD26/DPP IV in chemokine processing. Chem. Immunol. 72, 42–56 (1999).

  21. 21

    Colantonio, L. et al. Upregulation of integrin α6/β1 and chemokine receptor CCR1 by interleukin-12 promotes the migration of human type 1 helper T cells. Blood 94, 2981–2989 (1999).

  22. 22

    Berlin, C. et al. α4β7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell 74, 185–195 (1993).

  23. 23

    Simon, A.K., Seipelt, E. & Sieper, J. Divergent T-cell cytokine patterns in inflammatory arthritis. Proc. Natl Acad. Sci. USA 91, 8562–8566 (1994).

  24. 24

    Dolhain, R.J.E.M., van der Heiden, A.N., ter Haar, N.T., Breedveld, F.C. & Miltenburg, A.M.M. Shift toward T lymphocytes with a T helper 1 cytokine-secretion profile in the joints of patients with rheumatoid arthritis. Arthritis Rheum. 39, 1961–1969 (1996).

  25. 25

    Lockhart, D.J. et al. Expression monitoring by hybridization to high-density oligonucleotide arrays. Nature Biotechnol. 14, 1675–1680 (1996).

  26. 26

    Fambrough, D., McClure, K., Kazlauskas, A. & Lander, E.S. Diverse signaling pathways activated by growth factor receptors induce broadly overlapping, rather than independent, sets of genes. Cell 97, 727–741 (1999).

  27. 27

    Verzeletti, S. et al. Herpes simplex virus thymidine kinase gene transfer for controlled graft-versus-host disease and graft-versus-leukemia: clinical follow-up and improved new vectors. Hum. Gene Ther. 9, 2243–2251 (1998).

  28. 28

    Coligan, J.E., Kruisbeek, A.M., Margulies, D.H., Shevach, E.M. & Strober, W. Current Protocols in Immunology (Greene & Wiley, New York, 1991).

  29. 29

    Eisen, M.B., Spellman, P.T., Brown, P.O. & Botstein, D. Cluster analysis and display of genome-wide expression patterns. Proc. Natl Acad. Sci. USA 95, 14863–14868 (1998).

Download references

Acknowledgements

We thank P. Vigano for cord blood samples, and J. Sninsky and L. Adorini for helpful discussions and critical reading of the manuscript.

Author information

Affiliations

  1. Roche Milano Ricerche, Milano, Italy

    • Lars Rogge
    • , Mauro Biffi
    • , Elisa Bono
    •  & Francesco Sinigaglia
  2. DIBIT, Istituto Scientifico San Raffaele, Milano, Italy

    • Elisabetta Bianchi
  3. Roche Molecular Systems, Alameda, California, USA

    • Sheng-Yung P. Chang
    • , Heather Alexander
    •  & Chris Santini
  4. TIGET-H. S. Raffaele, Milano, Italy

    • Giuliana Ferrari
  5. Chair of Rheumatology, Istituto Ortopedico Gaetano Pini, Milano, Italy

    • Luigi Sinigaglia
  6. Roche Genetics, Basel, Switzerland

    • Monika Seiler
    • , Jan Mous
    •  & Ulrich Certa
  7. PRPZ-B, F. Hoffmann-La Roche Ltd, Basel, Switzerland

    • Martin Neeb

Authors

  1. Search for Lars Rogge in:

  2. Search for Elisabetta Bianchi in:

  3. Search for Mauro Biffi in:

  4. Search for Elisa Bono in:

  5. Search for Sheng-Yung P. Chang in:

  6. Search for Heather Alexander in:

  7. Search for Chris Santini in:

  8. Search for Giuliana Ferrari in:

  9. Search for Luigi Sinigaglia in:

  10. Search for Monika Seiler in:

  11. Search for Martin Neeb in:

  12. Search for Jan Mous in:

  13. Search for Francesco Sinigaglia in:

  14. Search for Ulrich Certa in:

Corresponding author

Correspondence to Lars Rogge.

Supplementary information

About this article

Publication history

Received

Accepted

Issue Date

DOI

https://doi.org/10.1038/75671

Further reading