MULTIPARAMETRIC IMMUNOPHENOTYPING OF B-CELLS IN PERIPHERAL-BLOOD OF HEALTHY-ADULTS BY FLOW-CYTOMETRY

The investigation of patients suffering from malignant lymphomas of th e B-cell type requires flow cytometric immunophenotyping, Several repo rts described the expression of almost all B lineage antigens on norma l and abnormal B lymphocytes. Thus, immunophenotyping of lymphomas mus t be interpreted in the context of the reference values obtained for h ealthy control individuals, For this purpose multiparametric Row cytom etric analysis offers the unique feature for lymphocyte subset analysi s. In the present study B lymphocytes in the peripheral blood of healt hy adults were investigated by multiparametric flow cytometric immunop henotyping for the detection of the frequency (in percent) of antigens provided by the revised European-American classification of lymphoid neoplasms (REAL) classification, Thus, 84 healthy adults were investig ated and grouped by age (average ages were as follows: group 1, 25.38 gears; group 2, 33.86 gears; group 3, 44.17 years; group 4, 55.67 gear s; group 5, 66.67 years). Analysis was done for surface immunoglobulin s (kappa and lambda chains of immunoglobulin M [IgM] and IgD) as well as CD10, CD11c, CD23, CD38, CD103, FMC-7, and B-B4. Three-color immuno phenotyping was performed for kappa/CD19/CD5, lambda/CDL9/C5)5, surfac e IgM/surface IgD/CD19, FMC-7/CD19/CD5, CD103/CD11c/CD19, CD10/CD23/CD 19, and CD38/B-B4/CD19 by live gating of CD19(+) events (n = 2,000), A lthough some numerical differences could be obtained for the different groups, statistical differences (P < 0.005) could only be obtained fo r the CD19(+)/CD5(+) B-cell subset, which was decreased in the elderly patients (group 5). The established two-color and three-color stainin gs will serve as a basis for future multiparametric immunophenotyping of abnormal lymphocytes (e.g,, for patients suffering from non-Hodgkin 's lymphoma of the B-cell type).