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Clinical Chemistry 28: 1905-1909, 1982;
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Clinical Chemistry, Vol 28, 1905-1909, Copyright © 1982 by American Association for Clinical Chemistry

Rapid enumeration of T lymphocytes by a flow-cytometric immunofluorescence method

SH Ip, CW Rittershaus, KW Healey, CC Struzziero, RA Hoffman and PW Hansen

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.


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