title: High-resolution analysis of T-cell receptor beta-chain repertoires using DNA heteroduplex tracking: generally stable, clonal CD8+ expansions in all healthy young adults.
abstract: The accurate measurement of T-cell receptor (TCR) repertoire changes requires the analysis of a representative sampling of complex T-cell populations. The number and frequency of clonally expanded TCR beta-chain transcripts bearing distinct CDR3 sequences were accurately determined using a simple DNA heteroduplex tracking assay. This method allowed major and minor clonal expansions (> or = 1% of a Vbeta subfamily's transcripts) to be rapidly and reproducibly quantified. Oligoclonal CD8 + cell expansions were detected in all young adults tested, while CD4 + cells generally expressed more polyclonal beta-chain repertoires. The same pattern of CD8 + cells oligoclonality and CD4 + cells polyclonality was observed in asymptomatic HIV-1 infected individuals with high CD4 + cell counts. CD8 + CD45RA + and CD8 + CD45RO + cell fractions both displayed oligoclonal, although distinct, TCR beta chain repertoires while CD8 + cells from umbilical cord blood were generally polyclonal. Oligoclonal CD8 + cell repertoires from young adults were generally stable over a period of weeks, although minor, transient, clonal expansions could also be detected in the absence of symptomatic infections. DNA heteroduplex tracking analysis provided a higher level of sensitivity for the detection of TCR beta chain transcript expansions than CDR3 length (spectrotyping/immunoscope) analysis. DNA heteroduplex tracking of TCR beta-chain transcripts is therefore a simple and sensitive method for assessing the level of clonality and for measuring changes in the TCR beta chain repertoire of different T-cell populations.
authors.affiliation: Aaron Diamond AIDS Research Center and The Rockefeller University, New York, NY 10016, USA.
authors.forename: D F
authors.initials: DF
authors.lastname: Shen
authors.forename: L
authors.initials: L
authors.lastname: Doukhan
authors.forename: S
authors.initials: S
authors.lastname: Kalams
authors.forename: E
authors.initials: E
authors.lastname: Delwart
mesh_terms: D000328:Adult; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000648:ultrastructure*; D002999:Clone Cells; D004247:DNA / Q000032:analysis* / Q000235:genetics; D015342:DNA Probes; D004277:DNA, Single-Stranded; D004586:Electrophoresis; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D008213:Lymphocyte Activation; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis*; D011110:Polymorphism, Genetic; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*
chemical_list: D015342:DNA Probes; D004277:DNA, Single-Stranded; D009692:Nucleic Acid Heteroduplexes; D016693:Receptors, Antigen, T-Cell, alpha-beta; D004247:DNA; D017493:Leukocyte Common Antigens