Myeloid and lymphoid chimerism after T-cell-depleted bone marrow transplantation: evaluation of conditioning regimens using the polymerase chain reaction to amplify human minisatellite regions of genomic DNA.
Myeloid and lymphoid chimerism after T-cell-depleted bone marrow transplantation: evaluation of conditioning regimens using the polymerase chain reaction to amplify human minisatellite regions of genomic DNA.
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DOI：
10.1182/blood.v80.12.3235.bloodjournal80123235
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发表时间：
1992-12
期刊：
影响因子：
20.3
通讯作者：
Stephen Mackinnon;L. Barnett;Jean Bourhis;P. Black;G. Heller;R. O'reilly
中科院分区：
文献类型：
--
作者：
Stephen Mackinnon;L. Barnett;Jean Bourhis;P. Black;G. Heller;R. O'reilly
测定t细胞枯竭异体骨髓移植（BMT）后骨髓和淋巴嵌合现象有助于了解移植生物学，并为评估不同调节方案促进移植的能力提供一种合理的方法。我们采用快速、灵敏的聚合酶链反应（PCR）方法评估骨髓和t细胞嵌合，前瞻性地研究了不同移植前调节方案在29例白血病患者骨髓移植后的作用。小卫星是DNA的高可变区域，由核心核苷酸序列的串联重复组成，等位基因多态性是由重复次数的差异引起的。我们使用这种变异来区分骨髓移植后的供体细胞和受体细胞。17名患者（9名兄弟姐妹和8名非亲属供体）接受了高分割全身照射（TBI）、硫替帕和环磷酰胺（Cy）的治疗。在其他12名患者（所有兄弟姐妹供体）中，11名患者接受TBI + Cy +另一种药物：VP16、碳铂或AZQ。1例患者接受TBI +硫替帕+ VP16治疗。除了一名患者外，所有接受研究的患者都接受了来自hla相同捐赠者的骨髓。PCR分析证实6例患者在移植后8天内有供体淋巴细胞植入。在移植后的前4周内，无论何种调理方案，所有的粒细胞DNA都来自供体。在17例接受TBI +硫替帕+ Cy治疗的患者中，14例患者的day +28 T细胞完全来自供体，但在12例接受其他治疗方案的患者中，10例患者的day +28 T细胞是混合嵌合的（P < 0.001）。1例不相关的TBI +硫替帕+ Cy的移植受者出现早期移植排斥反应。12例混合t细胞嵌合患者中有3例出现晚期移植失败，16例完全供体嵌合患者中没有一例出现晚期移植失败。然而，在第28天完全t细胞嵌合的16例患者中，有5例发生了急性移植物抗宿主病（GVHD），而混合嵌合的患者没有发生急性GVHD。我们的研究结果表明，微卫星PCR是一种快速而敏感的评估bmt后嵌合的方法，供体T细胞对于持续持久的移植很重要，TBI + thiotepa + Cy在诱导完全供体嵌合方面可能优于其他研究方案。需要更大的数据和更长时间的随访来证实这些数据，并评估完全供体t细胞嵌合与无白血病生存之间的关系。
Determining both myeloid and lymphoid chimerism after T-cell-depleted allogeneic bone marrow transplantation (BMT) could be helpful in the understanding of the biology of engraftment and could provide a rational method of assessing the ability of different conditioning regimens to promote engraftment. We prospectively investigated the role of different pretransplant conditioning regimens in 29 leukemic patients post-BMT by assessing myeloid and T-cell chimerism using a rapid and sensitive polymerase chain reaction (PCR) method. Minisatellites are hypervariable regions of DNA consisting of tandem repeats of a core nucleotide sequence, and allelic polymorphism results from differences in the number of the repeats. We used this variation to distinguish between donor and recipient cells post-BMT. Seventeen patients (9 sibling and 8 unrelated donors) received conditioning with hyperfractionated total body irradiation (TBI), thiotepa, and cyclophosphamide (Cy). Of the other 12 patients (all sibling donors), 11 received TBI plus Cy plus another agent: VP16, carboplatinum, or AZQ. One patient received TBI plus thiotepa plus VP16. All but one of the patients studied received marrow from HLA-identical donors. PCR analysis confirmed donor lymphoid engraftment within 8 days of transplant in six of six patients studied. All granulocyte DNA was of donor origin within the first 4 weeks of transplant, regardless of the conditioning regimen. The day +28 T cells were exclusively of donor origin in 14 of 17 patients who received TBI plus thiotepa plus Cy, but were mixed chimeric in 10 of 12 patients who received other conditioning regimens (P < .001). Early graft rejection was seen in one unrelated transplant recipient conditioned with TBI plus thiotepa plus Cy. Late graft failure was observed in 3 of 12 patients with mixed T-cell chimerism and in none of 16 patients with full donor chimerism at day +28. However, 5 of 16 patients who had complete T-cell chimerism at day +28 developed acute graft-versus-host disease (GVHD), whereas no patient with mixed chimerism had acute GVHD. Our results indicate that minisatellite PCR is a rapid and sensitive method for assessing chimerism post-BMT, that the donor T cells are important for consistent durable engraftment, and that TBI plus thiotepa plus Cy may be superior to the other regimens studied in inducing full donor chimerism. Larger numbers and longer follow-up are necessary to confirm these data and also to assess the relationship between complete donor T-cell chimerism and leukemia-free survival.