Martinelli G, Terragna C, Lemoli R M, Cavo M, Benni M, Motta M R, Amabile M, Ottaviani E, Testoni N, de Vivo A, Tura S
Institute of Hematology and Medical Oncology "Seràgnoli", University of Bologna, Italy.
Haematologica. 1999 May;84(5):397-404.
Autologous blood stem cell transplantation (ABSCT) using chemotherapy-induced mobilization of peripheral blood stem cells (PBSC) is being increasingly used in the treatment of multiple myeloma (MM). We report the clinical and molecular follow-up of 10 MM patients who underwent autologous stem cell transplantation with peripheral blood selected CD34+ cells, as support therapy following a myeloablative conditioning regimen.
The CDR-III coding region of the IgH gene was studied by a) consensus PCR applied to 8 MM patients, or b) by direct sequencing of PCR product generated by family-specific primers in the remaining two patients (who became immunofixation analysis (IF) negative). In this case, two patient-specific primers were generated, thus obtaining a high PCR assay sensitivity and specificity (ASO PCR).
Seven patients are alive: 4 of them have serum M protein assessable by IF, while 1 was not a secretor and 2 converted from serum IF positivity to negativity 6 and 12 months after ABSCT. Three patients died: 1 from disease progression and 2 from infective complications during clinical remission. The molecular analysis during the follow-up showed that the bone marrow samples from the two patients who obtained IF negativity were persistently PCR positive for the presence of rearranged CDR-III region. Moreover, despite the remarkable reduction of myeloma burden, a minimal level of residual myeloma cells was still detectable by molecular analysis.
These results confirm that although positive selection of CD34+ cells markedly reduces the contamination of myeloma cells from apheresis products by up to 3 log, and provides a cell suspension capable of restoring normal hematopoiesis after ablative conditioning regimen, it does not abrogate myeloma cell contamination in most of the apheresis products.
利用化疗诱导外周血干细胞（PBSC）动员进行的自体血干细胞移植（ABSCT）在多发性骨髓瘤（MM）治疗中的应用日益广泛。我们报告了10例MM患者的临床及分子随访情况，这些患者在接受清髓性预处理方案后，接受了外周血选择的CD34 +细胞自体干细胞移植作为支持治疗。
通过以下方法研究IgH基因的CDR-III编码区：a）对8例MM患者应用共识PCR，或b）对其余2例患者（免疫固定分析（IF）转为阴性）使用家族特异性引物生成的PCR产物进行直接测序。在这种情况下，生成了两个患者特异性引物，从而获得了高灵敏度和特异性的PCR检测方法（等位基因特异性寡核苷酸PCR）。
7例患者存活：其中4例可通过IF检测到血清M蛋白，1例不是分泌型，2例在ABSCT后6个月和12个月从血清IF阳性转为阴性。3例患者死亡：1例死于疾病进展，2例死于临床缓解期的感染并发症。随访期间的分子分析表明，2例IF转为阴性的患者骨髓样本中，重排的CDR-III区域持续呈PCR阳性。此外，尽管骨髓瘤负担显著减轻，但通过分子分析仍可检测到最低水平的残留骨髓瘤细胞。
这些结果证实，尽管CD34 +细胞的阳性选择可使单采产品中骨髓瘤细胞的污染显著减少多达3个对数，并提供一种能够在清髓性预处理方案后恢复正常造血的细胞悬液，但在大多数单采产品中并不能消除骨髓瘤细胞的污染。
