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The Journal of Immunology, 2005, 174: 1513-1524.
Copyright © 2005 by The American Association of Immunologists

Functional and Molecular Analysis of the Double-Positive Stage-Specific CD8 Enhancer E8III during Thymocyte Development1

Nicholas Feik*, Ivan Bilic*, Johanna Tinhofer2,*, Bernd Unger*,{dagger}, Dan R. Littman{ddagger} and Wilfried Ellmeier3,*

* Institute of Immunology, Medical University Vienna, and {dagger} Competence Center for Biomolecular Therapeutics, Vienna, Austria; and {ddagger} Molecular Pathogenesis Program, Skirball Institute of Biomolecular Medicine, Howard Hughes Medical Institute, New York University School of Medicine, New York, NY 10016

Several developmental stage-, subset-, and lineage-specific Cd8 cis-regulatory regions have been identified. These include the E8III enhancer, which directs expression in double-positive (DP) thymocytes, and E8II, which is active in DP cells and CD8+ T cells. Using a transgenic reporter expression assay, we identified a 285-bp core fragment of the E8III enhancer that retains activity in DP thymocytes. In vitro characterization of the core enhancer revealed five regulatory elements that are required for full enhancer activity, suggesting that multiple factors contribute to the developmental stage-specific activity. Furthermore, deletion of E8III in the mouse germline showed that this enhancer is required for nonvariegated expression of CD8 in DP thymocytes when E8II is also deleted. These results indicate that E8III is one of the cis-elements that contribute to the activation of the Cd8a and Cd8b gene complex during T cell development.


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The JI 2005 174: 1139-1141. [Full Text]  





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