Rapid enumeration of T lymphocytes by a flow-cytometric immunofluorescence method.
Combined with current advances in microprocessor-controlled flow cytometers, monoclonal antibodies provide a rapid means of phenotyping individual cell surface markers for a large number of clinical samples accurately and reproducibly, which may provide useful information in diagnosing disease and monitoring patients. We have developed a one-step flow-cytometric immunofluorescence procedure for enumerating E-rosette lymphocytes from whole blood by using the monoclonal antibody OKT11. This antibody recognizes the sheep erythrocyte receptor on the lymphocyte surface and can block sheep E-rosette formation. The flow cytometer we use, an Ortho Spectrum III, distinguishes lymphocytes from other leukocytes by measuring the narrow forward and right-angle light-scattering properties of the cells. The instrument further differentiates T lymphocytes frm non-T lymphocytes by measuring the green fluorescence signal of the OKT11-positive lymphocytes. In a typical sample, 1500--2500 lymphocytes are counted in 25 s. In a study of 158 patient samples, ranging from 1% to greater than 90% E-rosette-positive lymphocytes, the correlation coefficient between the manual E-rosette count and the flow immunofluorescence measurement is 0.943.
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D002452:Cell Count; D002462:Cell Membrane / Q000276:immunology; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007834:Lasers; D012397:Rosette Formation; D013601:T-Lymphocytes / Q000032:analysis / Q000166:cytology* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface