非霍奇金淋巴瘤自体外周血干细胞中肿瘤污染的临床研究

Contamination with minimal residual disease in autologous peripheral stem cell collected from non-Hodgkin′s lymphoma patients treated with high-dose therapy

目的 了解非霍奇金淋巴瘤 (NHL)患者自体外周血干细胞 (APBSC)移植物中肿瘤细胞污染的情况 ,探讨移植物被肿瘤细胞污染对预后的影响。方法 以免疫球蛋白重链 (IgH)、T细胞受体γ(TCRγ)基因克隆性重排和BCL 2 /IgH融合基因为检测标志 ,应用PCR结合SSCP技术检测了 33例NHL患者APBSC标本中的微量肿瘤细胞 ,同时还检测了部分患者自体外周血干细胞移植 (APBSCT)前后骨髓中的微量肿瘤细胞。结果 8例 (2 4 .2 % )患者的 17份 (2 3.6 % )APBSC标本微量肿瘤细胞检测阳性。动员时骨髓微量肿瘤细胞检测为阳性和阴性的患者 ,其APBSC污染阳性率分别为 6 6 .7%(6 / 9)和 8.7% (2 / 2 3) ,二者差异有显著性 (P <0 .0 1)。APBSC污染阳性和阴性患者的移植后预期 3年总生存率分别为 71.4 %和 71.2 % ,预期 3年无病生存率分别为 2 5 .0 %和 6 1.5 %。结论 NHL患者采集的APBSC可受到肿瘤细胞的污染。APBSC动员时骨髓微量肿瘤细胞阳性增加APBSC污染的几率。

Objective To detect the contaminating minimal residual disease (MRD) in autologous peripheral blood stem cells (APBSC) and evaluate its impact on the prognosis of non Hodgkin′s lymphoma (NHL) patients. Methods Minimal residual disease was detected in 72 APBSC samples from 33 NHL patients through PCR or PCR combined with DNA single strand conformation polymorphism analysis (SSCP) with the BCL 2/IgH, clonal rearrangement of IgH and TCR γ gene as markers. Minimal residual disease was also monitored in bone marrow samples collected pre , post induction chemotherapy and post transplantation. Results MRD was positive in 17/72 (23.6%) APBSC samples. The incidence of positive MRD in bone marrow pre , post induction chemotherapy and post transplantation was 44.0% (11/25), 28.1% (9/32) and 11.5% (3/26) respectively. Six (66.6%) of 9 patients with positive MRD in pre mobilization bone marrow, compared with 2 (8.7%) of 23 patients with negative MRD in bone marrow, were positive in contamination ( P <0.01). The estimated overall 3 year post transplantation survival rate for patients with positive and negative MRD in their APBSC would be 71.4% and 71.2% respectively, and the estimated 3 year disease free survival rates were 25.0% and 61.5% respectively ( P = 0.53 ). Conclusion APBSC collected from NHL patients after mobilization by chemotherapy combined with colony stimulating factor may be contaminated by lymphoma cells. The presence of minimal residual disease in bone marrow at mobilization may increase the incidence of APBSC contamination.