McDaniel H B, Yang M, Sidner R A, Jindal R M, Sahota A
Department of Surgery, Indiana University Hospital, Indianapolis 46202-5250, USA.
Clin Transplant. 1999 Apr;13(2):187-92. doi: 10.1034/j.1399-0012.1999.130206.x.
We evaluated peripheral blood microchimerism in 48 consecutive organ transplant recipients (35 kidneys, ten livers, one kidney-liver, one kidney-pancreatic islet, one kidney pancreas) up to 12 months post-transplantation. Patients were categorized according to the presence or absence of rejection episodes, and the patterns of microchimerism in the two groups were then compared.
DNA was extracted from donor, pre-transplant, and post-transplant peripheral blood samples. Several polymerase chain reaction (PCR)-based assays were developed for the detection of microchimerism. Assay sensitivities ranged from 0.0001 to 3%.
Microchimerism was detected only in sex-mismatched cases (male donors and female recipients) using nested PCR for a Y-chromosome marker. There were ten such cases (six kidneys, two livers, and two combined organ transplants). In patients without rejection (n = 7), there was a peak of donor-DNA at 1-3 wk post-transplantation followed by a second peak between 3 wk and 4 months. In patients with biopsy-proven rejection (n = 3), the peaks were absent and the levels of microchimerism were extremely low (< 0.001%). Microchimerism levels declined in all 10 patients and were barely detectable 1 yr post-transplantation. Microchimerism was not detected in the remaining 38 patients despite using a battery of sensitive PCR-based assays.
In our study, microchimerism was detected using the Y-chromosome PCR assay only and the level of donor-DNA in a given patient varied over time. This study highlights the difficulties in establishing a correlation between microchimerism and transplant tolerance.
我们评估了48例连续的器官移植受者(35例肾移植、10例肝移植、1例肝肾联合移植、1例肾-胰岛移植、1例肾-胰腺移植)移植后12个月内的外周血微嵌合体情况。根据是否发生排斥反应对患者进行分类,然后比较两组的微嵌合体模式。
从供体、移植前和移植后的外周血样本中提取DNA。开发了几种基于聚合酶链反应(PCR)的检测方法来检测微嵌合体。检测灵敏度范围为0.0001%至3%。
仅在使用巢式PCR检测Y染色体标记物的性别不匹配病例(男性供体和女性受者)中检测到微嵌合体。有10例此类病例(6例肾移植、2例肝移植和2例联合器官移植)。在无排斥反应的患者(n = 7)中,移植后1 - 3周出现供体DNA峰值,随后在3周和4个月之间出现第二个峰值。在经活检证实有排斥反应的患者(n = 3)中,未出现峰值,微嵌合体水平极低(< 0.001%)。所有10例患者的微嵌合体水平均下降,移植后1年几乎检测不到。尽管使用了一系列基于PCR的灵敏检测方法,但在其余38例患者中未检测到微嵌合体。
在我们的研究中,仅使用Y染色体PCR检测法检测到了微嵌合体,且给定患者体内的供体DNA水平随时间变化。本研究凸显了在建立微嵌合体与移植耐受之间相关性方面的困难。