title: Quantitative flow cytometry for the analysis of T cell receptor Vbeta chain expression.
abstract: Detailed characterisation of the T cell receptor (TCR) repertoire expressed by peripheral blood lymphocytes has been used to study specific T cell responses in disease conditions. The methods have mostly involved molecular biology analysis of transcribed gene products isolated from T cell subsets or individual clones. Extensive characterisation of the TCR Vbeta chain repertoire by flow cytometry is now possible due to the recently increased availability of specific monoclonal antibodies. However, there are major logistical problems inherent in this analysis relating to the number of cells required to obtain accurate results and the vast amounts of data generated. To reduce these factors to a practical level, we have performed a detailed study to define the limits of precision of cell subset analysis by flow cytometry. Maximal achievable precision was obtained by analysing 10(4) lymphocytes; no significant improvement was obtained by analysing greater numbers of cells up to 10(5) cells, even for cell subsets present at frequencies as low as 0.5%. Careful application of these precision profiles will also permit more effective use of clinical research samples for flow cytometry when the availability of cells is limited.
authors.affiliation: Division of Immunity and Infection, Rheumatology Research Group, University of Birmingham, UK.
authors.forename: J M
authors.initials: JM
authors.lastname: Faint
authors.forename: D
authors.initials: D
authors.lastname: Pilling
authors.forename: A N
authors.initials: AN
authors.lastname: Akbar
authors.forename: G D
authors.initials: GD
authors.lastname: Kitas
authors.forename: P A
authors.initials: PA
authors.lastname: Bacon
authors.forename: M
authors.initials: M
authors.lastname: Salmon
mesh_terms: D000293:Adolescent; D000328:Adult; D005434:Flow Cytometry; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000097:blood*; D012680:Sensitivity and Specificity; D016176:T-Lymphocyte Subsets / Q000737:chemistry
chemical_list: D007135:Immunoglobulin Variable Region; D016693:Receptors, Antigen, T-Cell, alpha-beta