2013
DOI: 10.1186/1471-2172-14-35
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Quantification of total T-cell receptor diversity by flow cytometry and spectratyping

Abstract: BackgroundT-cell receptor diversity correlates with immune competency and is of particular interest in patients undergoing immune reconstitution. Spectratyping generates data about T-cell receptor CDR3 length distribution for each BV gene but is technically complex. Flow cytometry can also be used to generate data about T-cell receptor BV gene usage, but its utility has not been compared to or tested in combination with spectratyping.ResultsUsing flow cytometry and spectratype data, we have defined a divergenc… Show more

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Cited by 24 publications
(17 citation statements)
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“…Previous analysis of specific virus specific CD8 cells have described both private and public (shared) clones between individuals (Klarenbeek et al, 2012; Dash et al, 2011). More global approaches to detecting and quantifying receptor diversity have used spectratyping to obtain a profile of CDR3 lengths, or flow cytometry to quantify V region usage (Ciupe et al, 2013; Pannetier et al, 1995; Faint et al, 1999; Lim et al, 2002). Both techniques have given interesting insights into clonal expansions associated with a variety of antigen-driven responses, although the sensitivity limits the detection of small clones.…”
Section: Discussionmentioning
confidence: 99%
“…Previous analysis of specific virus specific CD8 cells have described both private and public (shared) clones between individuals (Klarenbeek et al, 2012; Dash et al, 2011). More global approaches to detecting and quantifying receptor diversity have used spectratyping to obtain a profile of CDR3 lengths, or flow cytometry to quantify V region usage (Ciupe et al, 2013; Pannetier et al, 1995; Faint et al, 1999; Lim et al, 2002). Both techniques have given interesting insights into clonal expansions associated with a variety of antigen-driven responses, although the sensitivity limits the detection of small clones.…”
Section: Discussionmentioning
confidence: 99%
“…Conventional techniques to define T cell repertoire (namely, Southern blotting, flow cytometry, and polymerase chain reaction [PCR]) can reliably detect only the most abundant clones in biologic samples, and do not adequately cover the genetic diversity of the TCR . Typically, these tests are only informative about broad patterns of TCR Vβ usage.…”
Section: Introductionmentioning
confidence: 99%
“…77,84 Once the total number of T cells, or of a subset defined by naïve or memory surface markers, is established, we may ask how many TCR clonotypes there are, in each subset, in humans, mice and other mammals. [85][86][87][88] Estimates of the number of different TCRs that could, in principle, be produced by VDJ gene rearrangement in the thymus [89][90][91][92] far exceed the total number of T cells in a mouse or human body. 75,93 With the latest developments in sequencing techniques, is a direct count of the number of clonotypes in a mammal feasible?…”
Section: How Many Tcr Clonot Ype S Are Maintained?mentioning
confidence: 99%