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Quantification of total T-cell receptor diversity by flow cytometry and spectratyping.

Publication ,  Journal Article
Ciupe, SM; Devlin, BH; Markert, ML; Kepler, TB
Published in: BMC Immunol
August 6, 2013

BACKGROUND: T-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. RESULTS: Using flow cytometry and spectratype data, we have defined a divergence metric that quantifies the deviation from normal of T-cell receptor repertoire. We have shown that the sample size is a sensitive parameter in the predicted flow divergence values, but not in the spectratype divergence values. We have derived two ways to correct for the measurement bias using mathematical and statistical approaches and have predicted a lower bound in the number of lymphocytes needed when using the divergence as a substitute for diversity. CONCLUSIONS: Using both flow cytometry and spectratyping of T-cells, we have defined the divergence measure as an indirect measure of T-cell receptor diversity. We have shown the dependence of the divergence measure on the sample size before it can be used to make predictions regarding the diversity of the T-cell receptor repertoire.

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Published In

BMC Immunol

DOI

EISSN

1471-2172

Publication Date

August 6, 2013

Volume

14

Start / End Page

35

Location

England

Related Subject Headings

  • Sample Size
  • Receptors, Antigen, T-Cell
  • Polymerase Chain Reaction
  • Immunology
  • Humans
  • Flow Cytometry
  • Confidence Intervals
  • Complementarity Determining Regions
  • CD4-Positive T-Lymphocytes
  • CD3 Complex
 

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Ciupe, S. M., Devlin, B. H., Markert, M. L., & Kepler, T. B. (2013). Quantification of total T-cell receptor diversity by flow cytometry and spectratyping. BMC Immunol, 14, 35. https://doi.org/10.1186/1471-2172-14-35
Ciupe, Stanca M., Blythe H. Devlin, Mary Louise Markert, and Thomas B. Kepler. “Quantification of total T-cell receptor diversity by flow cytometry and spectratyping.BMC Immunol 14 (August 6, 2013): 35. https://doi.org/10.1186/1471-2172-14-35.
Ciupe SM, Devlin BH, Markert ML, Kepler TB. Quantification of total T-cell receptor diversity by flow cytometry and spectratyping. BMC Immunol. 2013 Aug 6;14:35.
Ciupe, Stanca M., et al. “Quantification of total T-cell receptor diversity by flow cytometry and spectratyping.BMC Immunol, vol. 14, Aug. 2013, p. 35. Pubmed, doi:10.1186/1471-2172-14-35.
Ciupe SM, Devlin BH, Markert ML, Kepler TB. Quantification of total T-cell receptor diversity by flow cytometry and spectratyping. BMC Immunol. 2013 Aug 6;14:35.
Journal cover image

Published In

BMC Immunol

DOI

EISSN

1471-2172

Publication Date

August 6, 2013

Volume

14

Start / End Page

35

Location

England

Related Subject Headings

  • Sample Size
  • Receptors, Antigen, T-Cell
  • Polymerase Chain Reaction
  • Immunology
  • Humans
  • Flow Cytometry
  • Confidence Intervals
  • Complementarity Determining Regions
  • CD4-Positive T-Lymphocytes
  • CD3 Complex