International Immunology, Vol. 11, No. 10, 1641-1650,
October 1999
© 1999 Japanese Society for Immunology
Unexpectedly late expression of intracellular CD3
and TCR 
proteins during adult thymus development
Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne,Ch. Des Boveresses 155, 1066 Epalinges, Switzerland
Correspondence to: A. Wilson
During adult thymus development immature CD4–CD8– [double-negative (DN)] precursor cells pass through four phenotypically distinct stages defined by expression of CD44 and CD25: CD44hiCD25– (DN1), CD44hiCD25+ (DN2), CD44loCD25+ (DN3) and CD44loCD25- (DN4). Although it is well established that the TCR ß, 
and 
genes are rearranged and expressed in association with the CD3 components in DN thymocytes, the precise timing of expression of the TCR and CD3 proteins has not been determined. In this report we have utilized a sensitive intracellular (ic) staining technique to analyze the expression of ic CD3
, TCR ß and TCR proteins in immature DN subsets. As expected from previous studies of TCR ß rearrangement and mRNA expression, icTCR ß+ cells were first detected in the DN3 subset and their proportion increased thereafter. Surprisingly, however, both icCD3+ and icTCR + cells were detected at later stages of development than was predicted by molecular studies. In particular icCD3 protein expression coincided with the transition from the DN2 to DN3 stage of development, whereas icTCR protein expression was only detected in a minor subset of DN4 cells. The implications of these findings for 
ß lineage divergence will be discussed.
Keywords: cellular differentiation, FACS, gene rearrangement, thymus
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