Tanaka J, Kasai M, Imamura M, Higa T, Sakurada K, Miyazaki T
3rd Department of Internal Medicine, Hokkaido University School of Medicine, Sapporo.
Rinsho Byori. 1994 Mar;42(3):249-56.
We assessed the origin of peripheral blood cells and bone marrow cells obtained from 15 patients after allogeneic bone marrow transplantation (allo BMT) by sensitive two-step polymerase chain reaction (PCR) amplification of MCT118, a variable number of tandem repeats regions (VNTR), that can be used to detect the DNA pattern of a minor cell population of only 1% without using radioisotopes. Mixed chimerism(MC) was detected in the haematopoietic cells of 3 patients. Two patients developed relapse of leukaemia after the detection of MC and one patient died of bone marrow hypoplasia 7 months after BMT. These findings indicate the clinical usefulness of this method to monitor patients with MC. Also, we analyzed cytokine gene expression in peripheral blood mononuclear cells during the development of graft-versus-host disease (GVHD) in patients who underwent allo BMT using a semiquantitative reverse-transcriptase polymerase chain reaction (RT-PCR). The expression of interleukin(IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha mRNA was increased during the development of GVHD and the degree of this increment depended on the severity of the disease. These findings suggest that IL-1 beta, IL-6, and TNF-alpha produced by peripheral blood mononuclear cells play an important role in the development of GVHD. Therefore, analysis of MC and cytokine mRNA expression using the PCR technique after allogeneic bone marrow transplantation provide important information for treatment and monitoring of marrow transplant patients.
我们通过对MCT118（一种可变数目串联重复序列区域，VNTR）进行灵敏的两步聚合酶链反应（PCR）扩增，评估了15例异基因骨髓移植（allo BMT）患者外周血细胞和骨髓细胞的来源。MCT118可用于在不使用放射性同位素的情况下检测仅占1%的少量细胞群体的DNA模式。在3例患者的造血细胞中检测到混合嵌合体（MC）。2例患者在检测到MC后发生白血病复发，1例患者在BMT后7个月死于骨髓发育不全。这些发现表明该方法在监测MC患者方面具有临床实用性。此外，我们使用半定量逆转录聚合酶链反应（RT-PCR）分析了接受allo BMT患者发生移植物抗宿主病（GVHD）期间外周血单个核细胞中的细胞因子基因表达。在GVHD发生期间，白细胞介素（IL）-1β、IL-6和肿瘤坏死因子（TNF）-α mRNA的表达增加，且这种增加的程度取决于疾病的严重程度。这些发现表明外周血单个核细胞产生的IL-1β、IL-6和TNF-α在GVHD的发生中起重要作用。因此，异基因骨髓移植后使用PCR技术分析MC和细胞因子mRNA表达可为骨髓移植患者的治疗和监测提供重要信息。
