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* Cellular Immunology Laboratory, Institut National de la Santé et de la Recherche Médical U543, Avenir Group, Hôpital Pitié-Salpêtrière, Université Pierre et Marie Curie-Paris6, Paris, France;
Multidisciplinary Oncology Center, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland;
Miltenyi Biotec, Cologne, Germany;
Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Lausanne Branch, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland; and
¶ Swiss Institute for Experimental Cancer Research, Epalinges, Switzerland
The differentiation of CD4+ or CD8+ T cells following priming of naive cells is central in the establishment of the immune response against pathogens or tumors. However, our understanding of this complex process and the significance of the multiple subsets of differentiation remains controversial. Gene expression profiling has opened new directions of investigation in immunobiology. Nonetheless, the need for substantial amount of biological material often limits its application range. In this study, we have developed procedures to perform microarray analysis on amplified cDNA from low numbers of cells, including primary T lymphocytes, and applied this technology to the study of CD4 and CD8 lineage differentiation. Gene expression profiling was performed on samples of 1000 cells from 10 different subpopulations, defining the major stages of post-thymic CD4+ or CD8+ T cell differentiation. Surprisingly, our data revealed that while CD4+ and CD8+ T cell gene expression programs diverge at early stages of differentiation, they become increasingly similar as cells reach a late differentiation stage. This suggests that functional heterogeneity between Ag experienced CD4+ and CD8+ T cells is more likely to be located early during post-thymic differentiation, and that late stages of differentiation may represent a common end in the development of T-lymphocytes.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This study was supported by Fond’action Contre le Cancer, Lausanne, the National Center of Competence in Research (NCCR) Molecular Oncology, Switzerland, the Nelia et Amadeo Barletta Foundation and the Institut National de la Santé et de la Recherche Médicale AVENIR Grant, France.
2 V.A. and A.B. participated equally in this work.
3 Address correspondence and reprint requests to Dr. Andreas Bosio, Miltenyi Biotec GmbH, Stöckheimer Weg 1, Cologne, Germany. E-mail address: andreas.bosio{at}miltenyibiotec.de
4 Abbreviations used in this paper: SAM, statistical analysis of microarrays.
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