Levy Y, Schmitt C, Tsapis A, Brouet J C, Fermand J P
Laboratory of Immunochemistry and Immunopathology, INSERM U 108, Hôpital Saint-Louis, Paris, France.
Clin Exp Immunol. 1991 Jun;84(3):435-9.
We have studied the phenotype and the immunoglobulin gene configuration of blood B cells from 15 patients with stage III multiple myeloma (MM) at diagnosis. Highly purified B cells (greater than 90% CD20 positive cells) were obtained after L-leucine methyl ester monocyte depletion and elimination of T cells by rosetting. The percentage of B cells with surface immunoglobulin (sIg) featuring the same light and heavy chain isotype as the serum monoclonal immunoglobulin was very low, except in one patient, in whom 25-30% of B cells displayed surface and cytoplasmic immunoglobulin (cIg) sharing idiotypic determinants with the serum monoclonal IgG kappa. In all cases but one the percentage of circulating plasma cells accounted for less than 2% of the enriched B cell preparations. In one patient purified B cell population contained 30% of plasma cells and the immunoglobulin gene study revealed a rearranged JH hybridizing fragment identical in bone marrow and blood B cell DNA samples. In the other 14 cases no rearranged fragment was detected although we used a technique allowing the detection of at least 2% clonal cells. The absence of clonal cells in the patient whose B cells contained a high percentage of cells featuring surface IgG molecules was confirmed on purified sIgG-positive cells. In addition CD20-positive cells from this patient did not contain gamma mRNA. Therefore the IgG molecules were clearly extrinsic. Although the existence of clonal B lymphocytes or of myeloma idiotype related B cells cannot be ruled out, they escape detection by sensitive genetic studies of immunoglobulin genes.
我们研究了15例诊断为III期多发性骨髓瘤(MM)患者血液B细胞的表型和免疫球蛋白基因构型。通过L-亮氨酸甲酯单核细胞清除和玫瑰花结法清除T细胞后,获得了高度纯化的B细胞(>90% CD20阳性细胞)。除1例患者外,表面免疫球蛋白(sIg)轻链和重链同种型与血清单克隆免疫球蛋白相同的B细胞百分比非常低,该例患者中25%-30%的B细胞表面和胞质免疫球蛋白(cIg)与血清单克隆IgG κ共享独特型决定簇。除1例患者外,所有病例中循环浆细胞的百分比占富集B细胞制剂的不到2%。1例患者的纯化B细胞群体中含有30%的浆细胞,免疫球蛋白基因研究显示骨髓和血液B细胞DNA样本中存在相同的重排JH杂交片段。在其他14例病例中,尽管我们使用了能够检测至少2%克隆细胞的技术,但未检测到重排片段。在B细胞中含有高百分比表面IgG分子的患者的纯化sIgG阳性细胞上证实不存在克隆细胞。此外,该患者的CD20阳性细胞不含γ mRNA。因此,IgG分子显然是外源性的。虽然不能排除克隆性B淋巴细胞或骨髓瘤独特型相关B细胞的存在,但它们通过免疫球蛋白基因的敏感遗传学研究无法检测到。