Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry

Abstract

The investigation of patients suffering from malignant lymphomas of the B-cell type requires flow cytometric immunophenotyping. Several reports described the expression of almost all B lineage antigens on normal and abnormal B lymphocytes. Thus, immunophenotyping of lymphomas must be interpreted in the context of the reference values obtained for healthy control individuals. For this purpose multiparametric flow cytometric analysis offers the unique feature for lymphocyte subset analysis. In the present study B lymphocytes in the peripheral blood of healthy adults were investigated by multiparametric flow cytometric immunophenotyping 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 investigated and grouped by age (average ages were as follows: group 1, 25.38 years; group 2, 33.86 years; group 3, 44.17 years; group 4, 55.67 years; group 5, 66.67 years). Analysis was done for surface immunoglobulins (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 immunophenotyping was performed for kappa/CD19/CD5, lambda/CD19/CD5, surface IgM/surface IgD/CD19, FMC-7/CD19/CD5, CD103/CD11c/CD19, CD10/CD23/CD19, and CD38/B-B4/CD19 by live gating of CD19+ events (n = 2,000). Although some numerical differences could be obtained for the different groups, statistical differences (P < 0.005) could only be obtained for the CD19+/CD5+ B-cell subset, which was decreased in the elderly patients (group 5). The established two-color and three-color stainings 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).

Spontaneous Phenotypic and Molecular Blood Remission in a Case of Chronic Lymphocytic Leukaemia

“…To our knowledge, we report the first case of spontaneous complete blood remission of CLL confirmed by the absence of phenotypic and molecular blood detection of the malignant clone, lasting almost 3 years. We detected < 4% of CD5/CD19 + cells, which corresponds to the value observed in normal individuals of the same age ( Höffkes et al , 1996 ). Persistence of phenotypic and molecular marrow detection of leukaemic cells suggests that long‐lasting remission may be observed even in the context of minimal residual disease.…”

Definition of leukocyte subsets in primate central nervous system by polychromatic flow cytometry

“…Macrophages in the CNS showed a shift to a higher CD11c expression level, whereas microglia cells were uniformly CD11c negative. CD11c is usually used to define myeloid DC (27, 42), but can also be found on B cell subsets (43). Therefore, inclusion of other lineage markers is necessary for proper analysis of these antigen presenting cells.…”

CD38 expression is an important prognostic marker in chronic lymphocytic leukaemia

“…Between May 1994 and December 2000, 133 consecutive patients with chronic lymphocytic leukaemia from a single institution were enrolled in this study and analysed for several biological and clinical characteristics: age, sex, Binet stage, white blood cell count, haemoglobin level, platelet count, lactate dehydrogenase, thymidine kinase, immunoglobulin A (IgA) serum-concentration, survival, treatment history and time from diagnosis to first treatment. In each patient morphologic diagnosis of B-CLL was confirmed flow cytometrically 14 revealing a typical CD19,20,5,23-positive Ig light chain, ( or light chain) restricted immunophenotype. Patient characteristics are shown in Table 1.…”