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The bcl-2 proto-oncogene, rearranged and deregulated in B-cell lymphomas bearing the t(14;18) translocation, encodes an inner mitochondrial membrane protein that blocks apoptotic cell death. We have developed a sensitive immunofluorescence assay for the single- and multicolor flow cytometric analysis of bcl-2 protein in relation to other markers and cell cycle, based on a fixation-permeation step of cells with paraformaldehyde and Triton X100 and the use of a bcl-2 specific monoclonal antibody (MoAb). As an application of this method, we have examined the expression of bcl-2 in normal and neoplastic lymphoid cells. We have found that greater than 80% of normal T-and B-cells are bcl-2 positive; following in vitro mitogen activation, the bcl-2 reactivity decreased slightly in the former but markedly in latter cells. In both cases the bcl-2 expression was not restricted to a specific phase of the cell cycle, as evidenced by two-color analysis. On lymphoblastoid cell lines, the bcl-2 staining intensity was variable and not necessarily correlated to molecular rearrangements of the bcl-2 gene. Among fresh B-cell non-Hodgkin's lymphomas (B-NHL), most sporadic Burkitt's cases were bcl-2 negative. Of four centroblastic-centrocytic cases with rearrangements of the bcl-2 gene, only two presented elevated amounts of bcl-2 protein, indicating that the levels of bcl-2 are not diagnostic of the translocation.(ABSTRACT TRUNCATED AT 250 WORDS)

Original publication

DOI

10.1002/cyto.990130509

Type

Journal article

Journal

Cytometry

Publication Date

1992

Volume

13

Pages

502 - 509

Keywords

Antibodies, Monoclonal, B-Lymphocytes, Cell Cycle, Flow Cytometry, Fluorescent Antibody Technique, Gene Expression Regulation, Neoplastic, Gene Rearrangement, Humans, Lymphocyte Activation, Lymphoma, Non-Hodgkin, Mitochondria, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-bcl-2, T-Lymphocytes, Translocation, Genetic, Tumor Cells, Cultured