HIGH-RESOLUTION ANALYSIS OF T-CELL RECEPTOR BETA-CHAIN REPERTOIRES USING DNA HETERODUPLEX TRACKING - GENERALLY STABLE, CLONAL CD8-YOUNG ADULTS( EXPANSIONS IN ALL HEALTHY)

The accurate measurement of T-cell receptor (TCR) repertoire changes r equires the analysis of a representative sampling of complex T-cell po pulations. The number and frequency of clonally expanded TCR beta-chai n transcripts bearing distinct CDR3 sequences were accurately determin ed using a simple DNA heteroduplex tracking assay. This method allowed major and minor clonal expansions(greater than or equal to 1% of a V beta subfamily's transcripts) to be rapidly and reproducibly quantifie d. Oligoclonal CD8 + cell expansions were detected in all young adults tested, while CD4 + cells generally expressed more polyclonal beta-ch ain repertoires. The same pattern of CD8 + cells oligoclonality and CD 4 + cells polyclonality was observed in asymptomatic HIV-1 infected in dividuals with high CD4 + cell counts. CD8 + CD45RA + and CD8 + CD45RO + cell fractions both displayed oligoclonal, although distinct, TCR b eta chain repertoires while CD8 + cells from umbilical cord blood were generally polyclonal. Oligoclonal CD8 + cell repertoires from young a dults were generally stable over a period of weeks, although minor, tr ansient, clonal expansions could also be detected in the absence of sy mptomatic infections. DNA heteroduplex tracking analysis provided a hi gher level of sensitivity for the detection of TCR beta chain transcri pt expansions than CDR3 length (spectrotyping/immunoscope) analysis. D NA heteroduplex tracking of TCR beta-chain transcripts is therefore a simple and sensitive method for assessing the level of clonality and f or measuring changes in the TCR beta chain repertoire of different T-c ell populations. (C) 1998 Published by Elsevier Science B.,V. All righ ts reserved.