Analysis and enumeration of T cells, B cells and NK cells using the monoclonal antibody fluorescence capability of a routine haematology analyser (Cell-Dyn CD4000).
This communication details a method for the quantitative and qualitative analysis of blood T-, B- and NK-cell populations using the Abbott Cell-Dyn CD4000 haematology analyser. A series of 66 ethylenediaminetetraacetic acid (EDTA)-anticoagulated samples with lymphocyte counts between 0.2 and 33.3 x 10(9)/l were selected and analysed with CD3, CD19, Ia and CD56 monoclonal reagents. The flow cytometry reference method utilized a lymphocyte gate defined by optical scatter, with phenotypic analyses referencing to this gate and the absolute lymphocyte count. The CD4000 method analysed all leucocyte events, set primary gates for specific immunophenotypic fractions, and then determined population counts by reference to the white blood cell (WBC) count. Comparisons of CD3+ T-cell and CD19+ B-cell numbers showed high coefficients of correlation (R(2) > 0.95) and agreement (y = 1.01x) between the CD4000 and flow cytometry reference methods. Lower coefficients of correlation were obtained for CD3-CD56+ (R(2) = 0.52) and CD3+CD56+ (R(2) = 0.83) components. No major discrepancies were observed, and the CD4000 procedures additionally provided qualitative insights into the possibility of T-cell activation. The potential to undertake immediate analysis of EDTA-anticoagulated blood samples to determine the nature of abnormal lymphocyte morphology or numbers represents a considerable advance in the capability of haematology laboratories.
Molero
T
T
Servicio de Hematologia y Hemoterapia, Hospital de Gran Canaria Dr Negrín, Barranco de la Ballena, Las Palmas, Gran Canaria, Spain.
Roemer
B
B
Perera Alvarez
M del Mar
Mdel M
Lemes
A
A
Iñigo
S de la Iglesia
Sde L
Palacios
G
G
Scott
C S
CS
D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D001322:Autoanalysis / Q000295:instrumentation / Q000379:methods; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002452:Cell Count; D005453:Fluorescence; D006403:Hematologic Tests / Q000295:instrumentation / Q000379:methods; D006801:Humans; D019044:Image Cytometry / Q000295:instrumentation* / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D016130:Immunophenotyping / Q000295:instrumentation / Q000379:methods; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology
D000911:Antibodies, Monoclonal