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Optimal gating strategies for determining bronchoalveolar lavage CD4/CD8 lymphocyte ratios by flow cytometry.
J Immunol Methods. 2004 Feb 01; 285(1):15-23.JI

Abstract

Alterations in bronchoalveolar lavage (BAL) CD4/CD8 T cell subset ratios have been demonstrated in a variety of different respiratory disorders and the measurement of these changes may be diagnostically helpful. Flow cytometry (FCM) is a precise technology that offers many advantages over conventional cytospin techniques to determine T cell subset ratios in tissue fluids such as BAL. However, the optimum gating strategies for evaluating these parameters by FCM have not been evaluated. Here, the CD4/CD8 ratios in 33 BAL samples were compared using three different methods by FCM with two different flow cytometers. Bland Altman analysis demonstrated clinically insignificant differences between two simplified staining and gating strategies and a more complex "gold standard" method. These findings confirm the precision of FCM for BAL T cell subset ratio analysis and suggest that the optimal gating strategy may be a simple panel using only CD45, CD4 and CD8.

Authors+Show Affiliations

HIV Immunology, Department of Clinical Immunology, Royal Free Hospital, Pond Street, London NW3 2QG, UK. sbarry@doctors.org.ukNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

14871531

Citation

Barry, Simon M., and George Janossy. "Optimal Gating Strategies for Determining Bronchoalveolar Lavage CD4/CD8 Lymphocyte Ratios By Flow Cytometry." Journal of Immunological Methods, vol. 285, no. 1, 2004, pp. 15-23.
Barry SM, Janossy G. Optimal gating strategies for determining bronchoalveolar lavage CD4/CD8 lymphocyte ratios by flow cytometry. J Immunol Methods. 2004;285(1):15-23.
Barry, S. M., & Janossy, G. (2004). Optimal gating strategies for determining bronchoalveolar lavage CD4/CD8 lymphocyte ratios by flow cytometry. Journal of Immunological Methods, 285(1), 15-23.
Barry SM, Janossy G. Optimal Gating Strategies for Determining Bronchoalveolar Lavage CD4/CD8 Lymphocyte Ratios By Flow Cytometry. J Immunol Methods. 2004 Feb 1;285(1):15-23. PubMed PMID: 14871531.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optimal gating strategies for determining bronchoalveolar lavage CD4/CD8 lymphocyte ratios by flow cytometry. AU - Barry,Simon M, AU - Janossy,George, PY - 2003/02/24/received PY - 2003/10/13/revised PY - 2003/10/27/accepted PY - 2004/2/12/pubmed PY - 2004/3/30/medline PY - 2004/2/12/entrez SP - 15 EP - 23 JF - Journal of immunological methods JO - J Immunol Methods VL - 285 IS - 1 N2 - Alterations in bronchoalveolar lavage (BAL) CD4/CD8 T cell subset ratios have been demonstrated in a variety of different respiratory disorders and the measurement of these changes may be diagnostically helpful. Flow cytometry (FCM) is a precise technology that offers many advantages over conventional cytospin techniques to determine T cell subset ratios in tissue fluids such as BAL. However, the optimum gating strategies for evaluating these parameters by FCM have not been evaluated. Here, the CD4/CD8 ratios in 33 BAL samples were compared using three different methods by FCM with two different flow cytometers. Bland Altman analysis demonstrated clinically insignificant differences between two simplified staining and gating strategies and a more complex "gold standard" method. These findings confirm the precision of FCM for BAL T cell subset ratio analysis and suggest that the optimal gating strategy may be a simple panel using only CD45, CD4 and CD8. SN - 0022-1759 UR - https://www.unboundmedicine.com/medline/citation/14871531/Optimal_gating_strategies_for_determining_bronchoalveolar_lavage_CD4/CD8_lymphocyte_ratios_by_flow_cytometry_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022175903004393 DB - PRIME DP - Unbound Medicine ER -