Liu Qi-fa, Li Yang-qiu, Yang Dong, Zhang Yu, Yang Li-jian, Chen Shao-hua, Sun Jing, Liu Xiao-li, Zhou Shu-yun
Department of Hematology, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Chin Med J (Engl). 2004 Mar;117(3):413-8.
We distinguished graft-versus-host disease (GVHD) from graft-versus-leukemia (GVL) effects and to investigate the distribution of T-cell receptor (TCR) V beta gene repertoire in individuals with leukemia before and after allogeneic hematopoietic stem cell transplantation (allo-HSCT).
Peripheral blood mononuclear cells (PBMC) were obtained from 10 normal individuals, 8 donors and 11 patients with leukemia before and after transplantation. Polymerase chain reaction (PCR) amplification of complementarity-determining region 3 (CDR3) of 24 TCR V beta genes was used to examine serial samples of PBMC. The PCR products were further analyzed by genescan to evaluate clonality of T cells.
The 24 TCR V beta gene repertoire displayed highly diverse and polyclonal spectratypes in all normal individuals and 4 of 8 donors. Another 4 donors expressed part of the 24 TCR V beta subfamily and 1 donor had oligoclonality. The expressions of the 24 TCR V beta subfamilies were skewed and restricted in 11 leukemia patients before and after transplantation. Some absences of 24 TCR V beta subfamily expression were quite similar between the recipients pro-transplantation and related donors. The number of subfamilies expressed increased over time post-transplantation, but the restricted expressions of the subfamily could last 6 - 30 months after transplantation. All patients with GVHD and some without GVHD exhibited T cell clonal expansion. The expansive T cell clone was distributed in V beta 2-3, 16-17, 18-19, 21 and V beta 23 in patients with GVHD and in V beta 7, 9, 16 and 19 in patients without GVHD. One patient with syngeneic-HSCT (syn-HSCT) had V beta 15 and 16 T cell expansion after transplantation. One patient displayed V beta 18 T cell expansion after donor lymphocyte infusion (DLI).
Normal individuals express the entire 24 TCR V beta gene repertoire and have polyclonal distribution. However, the TCR V beta gene repertoire is only partially expressed in some donors. The TCR V beta gene repertoire is restrictedly expressed in a skew fashion in patients with leukemia before and after transplantation. The number of TCR V beta gene subfamilies increases over time post-transplantation. GVHD and GVL effects may induce the proliferation of T cell clones. Clinical GVL response may be distinguished from GVHD alloreactivity through the host MHC antigen.
我们区分移植物抗宿主病（GVHD）和移植物抗白血病（GVL）效应，并研究异基因造血干细胞移植（allo-HSCT）前后白血病患者T细胞受体（TCR）Vβ基因谱的分布。
从10名正常个体、8名供者以及11名白血病患者移植前后获取外周血单个核细胞（PBMC）。采用聚合酶链反应（PCR）扩增24种TCR Vβ基因的互补决定区3（CDR3），用于检测PBMC的系列样本。PCR产物进一步通过基因扫描分析以评估T细胞的克隆性。
24种TCR Vβ基因谱在所有正常个体及8名供者中的4名中表现出高度多样和多克隆的谱型。另外4名供者表达了24种TCR Vβ亚家族的部分成员，1名供者具有寡克隆性。24种TCR Vβ亚家族的表达在11名白血病患者移植前后呈偏态且受限。移植前受者与相关供者之间24种TCR Vβ亚家族表达的一些缺失相当相似。移植后表达的亚家族数量随时间增加，但亚家族的受限表达在移植后可持续6至30个月。所有发生GVHD的患者及部分未发生GVHD的患者均表现出T细胞克隆性扩增。发生GVHD的患者中，扩增的T细胞克隆分布于Vβ2-3、16-17、18-19、21和Vβ23，未发生GVHD的患者中分布于Vβ7、9、16和19。1名接受同基因造血干细胞移植（syn-HSCT）的患者移植后出现Vβ15和16 T细胞扩增。1名患者在供者淋巴细胞输注（DLI）后出现Vβ18 T细胞扩增。
正常个体表达完整的24种TCR Vβ基因谱且呈多克隆分布。然而，TCR Vβ基因谱在一些供者中仅部分表达。TCR Vβ基因谱在白血病患者移植前后呈偏态受限表达。移植后TCR Vβ基因亚家族数量随时间增加。GVHD和GVL效应可能诱导T细胞克隆增殖。临床GVL反应可通过宿主MHC抗原与GVHD同种异体反应性相区分。
