Montecucco C, Riccardi A, Mazzini G, Danova M, Ucci G, Giordano P A
Basic Appl Histochem. 1985;29(3):275-82.
A double fluorescence assay has been employed for the detection of cell surface and/or cytoplasmic immunoglobulins (Ig) and the measurement of nuclear DNA content in the same cell. Following staining for Ig by means of FITC conjugated antibodies directed against heavy or light chains, cell suspensions or cytospin preparations were ethanol fixed and stained with a propidium iodide-RNAse solution. In this way, the cytometric DNA content of circulating B-lymphocytes was analyzed in three patients suffering from plasma cell malignancies with an excess of peripheral blood B-lymphocytes and evidence of aneuploid bone marrow plasma cells. Aneuploid circulating B-lymphocytes with the same DNA stem-line as bone marrow plasma cells were found in two patients with advanced disease but not in the only one we studied at presentation. Aneuploid lymphocytes had surface immunoglobulins bearing the same light chain as the M-protein. In addition, a significant percentage (23%) of cells lacking either surface or cytoplasmic immunoglobulins proved to be aneuploid in plasma cell leukemia. Nuclear DNA measurement combined with surface or cytoplasmic marker analysis appears to be a reliable method for studying neoplastic lymphoid precursor cells in plasma cell malignancies.
已采用双荧光测定法来检测细胞表面和/或细胞质免疫球蛋白(Ig),并测量同一细胞中的核DNA含量。用针对重链或轻链的异硫氰酸荧光素(FITC)偶联抗体对Ig进行染色后,将细胞悬液或细胞涂片制备物用乙醇固定,并用碘化丙啶-核糖核酸酶溶液染色。通过这种方式,对三名患有浆细胞恶性肿瘤且外周血B淋巴细胞增多并有非整倍体骨髓浆细胞证据的患者的循环B淋巴细胞的细胞计量DNA含量进行了分析。在两名晚期疾病患者中发现了与骨髓浆细胞具有相同DNA主干系的非整倍体循环B淋巴细胞,但在我们研究的唯一一名初诊患者中未发现。非整倍体淋巴细胞的表面免疫球蛋白带有与M蛋白相同的轻链。此外,在浆细胞白血病中,相当比例(23%)缺乏表面或细胞质免疫球蛋白的细胞被证明是非整倍体。核DNA测量与表面或细胞质标记分析相结合似乎是研究浆细胞恶性肿瘤中肿瘤性淋巴样前体细胞的可靠方法。