Brown R, Luo X F, Gibson J, Morley A, Sykes P, Brisco M, Joshua D
Institute of Haematology, Royal Prince Alfred Hospital, Sydney, Australia.
Br J Haematol. 1995 May;90(1):113-8. doi: 10.1111/j.1365-2141.1995.tb03388.x.
An mRNA in situ hybridization (ISH) method which used non-radioactive idiotypic oligonucleotide probes has been used to detect malignant cells in the bone marrow and peripheral blood of patients with multiple myeloma. For each of two patients with multiple myeloma a pair of biotinylated antisense oligonucleotide probes (18-22mer) was prepared from non-germline sequences of the rear-ranged immunoglobulin heavy chain (IgH) gene. These oligonucleotide sequences were not homologous with any previously published sequence. The probes from each patient were specific as shown by a failure to hybridize to any cells from six other myeloma patients and four normal individuals. Specific staining of IgH gene mRNA occurred only when the myeloma cells and the sequence of the probe used were from the same patient. Using simultaneous fluorescent immunocytochemistry it was shown that more than 95% of the ISH-positive cells expressed the malignant light chain in their cytoplasm. ISH positive cells were found in 1-4% of the peripheral blood mononuclear fraction of these two patients. These studies show that idiotypic oligonucleotide IgH gene probes can be used to identify individual cells belonging to the malignant clone and offer the possibility of developing innovative tumour-specific therapeutic procedures using antisense technology for patients with myeloma.
一种使用非放射性独特型寡核苷酸探针的mRNA原位杂交（ISH）方法已被用于检测多发性骨髓瘤患者骨髓和外周血中的恶性细胞。对于两名多发性骨髓瘤患者，从重排的免疫球蛋白重链（IgH）基因的非种系序列制备了一对生物素化的反义寡核苷酸探针（18 - 22聚体）。这些寡核苷酸序列与任何先前发表的序列均无同源性。如未能与其他六名骨髓瘤患者和四名正常个体的任何细胞杂交所示，来自每名患者的探针具有特异性。仅当骨髓瘤细胞与所用探针的序列来自同一患者时，才会出现IgH基因mRNA的特异性染色。使用同步荧光免疫细胞化学表明，超过95%的ISH阳性细胞在其细胞质中表达恶性轻链。在这两名患者外周血单核细胞部分的1 - 4%中发现了ISH阳性细胞。这些研究表明，独特型寡核苷酸IgH基因探针可用于识别属于恶性克隆的单个细胞，并为使用反义技术为骨髓瘤患者开发创新的肿瘤特异性治疗方法提供了可能性。
