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By Frank J. Dixon

The Scripps examine Institute, l. a. Jolla, CA. most modern findings within the box of immunology. converted define structure.

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1993). , 1995). Thus, it appears that rearrangements at the TCRa locus continue on both alleles until a receptor is formed that can bind to thymic MHC molecules. , 1995). , 1997) (see Section 111,B). Therefore, functional TCRP chains relay a negative feedback to the second allele in the absence of TCRa, strongly suggesting a role of the pre-TCR in mediating allelic exclusion. If the pre-TCR were implicated in the arrest of further TCRP rearrangements, pTa-deficient mice should show signs of inefficient allelic exclusion, a hypothesis that has been tested in three experimental approaches.

1997). Together, these results clearly demonstrated that the pTa chain and thus the pre-TCR were involved in allelic exclusion of endogenous VP rearrangements, at least in TCRPtransgenic mice. This conclusion has recently been extended to nontransgenic mice by comparing the rearrangement status of TCRP alleles in sorted T N thymocytes from pTa-deficient mice and wild-type CD25+CD44-"OW littermates at the single cell level. Using a set of JP2-specific and several VP-specific primers and the single-cell PCR technique, V(D)Jjoints were amplified from a number of pre-T cells exhibiting VP + (D)JP rearrange- three produce productive joints.

The efficiency of allelic exclusion in the absence of pTa was determined by measuring the frequency of mature SP thymocytes and peripheral LN T cells with two distinct TCRP chains on the cell surface in both pTa-’- and pTa’ mice. To calculate the maximum number of cells expressing two distinct TCRP chains in the complete absence of allelic exclusion, the following assumptions were made: (1)Sufficient time for rearrangements on both alleles, (2)absolutely no feedback inhibition of the functional P chain on the other allele, and (3) no selective disadvantage of cells with two functional /3 chains.

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