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外周血干细胞采集物中肿瘤污染的分子检测

Molecular detection of tumor contamination in peripheral blood stem cell harvests.

作者信息

Craig J I, Langlands K, Parker A C, Anthony R S

机构信息

Department of Haematology, Royal Infirmary of Edinburgh, Scotland, UK.

出版信息

Exp Hematol. 1994 Aug;22(9):898-902.

PMID:7914873
Abstract

A total of 96 peripheral blood stem cell harvests (PBSCH) were collected following standard chemotherapy from 27 patients with leukemia, lymphoma, and myeloma. Tumor molecular markers were analyzed in diagnostic samples by Southern blot [immunoglobulin heavy chain (IgHJ), T cell receptor (TcR) delta chain, TcR beta chain] and polymerase chain reaction (PCR) amplification [IgH, TcR delta, t(14;18) translocation] and found in 11 of 22 and 13 of 27 patients, respectively. At a sensitivity of 2 to 5%, Southern blot analysis failed to detect tumor in PBSCH. Using PCR with sensitivities of 10(-4) (IgH, TcR delta) to 10(-6) [t(14;18)] tumor was present in 34 PBSCH collected from five patients with acute lymphoblastic leukemia and two with lymphoma. In these patients, PBSCH collected after successive courses of chemotherapy remained consistently positive. However, no tumor was detected in 28 PBSCH from five patients with lymphoma and one with myeloma. Of eight patients who had bone marrow examined (six concurrently) within 3 weeks of PBSCH, one had tumor in the PBSCH but not in the bone marrow, and three had tumor in the bone marrow but not in the PBSCH, indicating a possible advantage in using PBSC for autologous transplantation in these patients. Although PBSC are an alternative source of stem cells to bone marrow and are considered to have a lower incidence of tumor contamination, the majority of PBSC in this study were positive by PCR analysis. PCR analysis was unable to determine if a positive result represents clonogenic cells capable of initiating relapse following transplant.

摘要

对27例白血病、淋巴瘤和骨髓瘤患者进行标准化疗后,共采集了96次外周血干细胞采集物(PBSCH)。通过Southern印迹法(免疫球蛋白重链(IgHJ)、T细胞受体(TcR)δ链、TcRβ链)和聚合酶链反应(PCR)扩增(IgH、TcRδ、t(14;18)易位)对诊断样本中的肿瘤分子标志物进行分析,分别在22例患者中的11例和27例患者中的13例中检测到。在灵敏度为2%至5%时,Southern印迹分析未能在PBSCH中检测到肿瘤。使用灵敏度为10(-4)(IgH、TcRδ)至10(-6) [t(14;18)]的PCR,在从5例急性淋巴细胞白血病患者和2例淋巴瘤患者采集的34份PBSCH中检测到肿瘤。在这些患者中,连续化疗疗程后采集的PBSCH始终呈阳性。然而,在5例淋巴瘤患者和1例骨髓瘤患者的28份PBSCH中未检测到肿瘤。在PBSCH采集后3周内对8例患者进行骨髓检查(其中6例同时进行),1例患者的PBSCH中有肿瘤但骨髓中没有,3例患者的骨髓中有肿瘤但PBSCH中没有,这表明在这些患者中使用外周血干细胞进行自体移植可能具有优势。尽管外周血干细胞是骨髓干细胞的替代来源,并且被认为肿瘤污染发生率较低,但本研究中的大多数外周血干细胞经PCR分析呈阳性。PCR分析无法确定阳性结果是否代表能够引发移植后复发的克隆形成细胞。

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