Molecular analysis of alloreactive CTL post-hemopoietic stem cell transplantation
Molecular analysis of alloreactive CTL post-hemopoietic stem cell transplantation
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DOI：
10.4049/jimmunol.179.3.2013
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发表时间：
2007-08-01
影响因子：
4.4
通讯作者：
Maciejewski, Jaroslaw P.
中科院分区：
文献类型：
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作者：
O'Keefe, Christine L.;Gondek, Lukasz;Maciejewski, Jaroslaw P.
移植物抗宿主病(GVHD)的诊断和监测实验室测试的发展因缺乏对触发同种异体反应的微小组织相容性AGS的了解而受到阻碍。我们推测，TCR独特的分子结构可以作为不明原因的AGS的标记，并可用于移植后GVHD的分子监测。为了鉴定同种异体反应性T细胞克隆，我们对一组接受造血干细胞移植的患者进行了体外同种异体刺激培养，并测定了免疫显性同种异体反应性克隆的CDR3序列；鉴定了10个相应的克隆型，仅限于活化的T细胞。作为鉴定同种异体反应克隆的另一种方法，分子TCR分析被应用于GVHD感染组织的活检。培养和活检获得的克隆型被用来设计序列特异性定量PCR分析，以监测移植后血液样本和随后的活检中假定的同种特异性克隆型的水平。由于用于鉴定免疫优势克隆类型的方法的合理设计，我们能够在移植过程中跟踪潜在的GVHD特异性T细胞的行为。根据我们的结果，我们得出结论，分子T细胞诊断可以成为监测移植后免疫反应的有力工具。
The development of laboratory tests for the diagnosis and monitoring of graft-vs-host disease (GVHD) is hampered by a lack of knowledge of minor histocompatibility Ags triggering alloresponses. We hypothesized that the unique molecular structure of the TCR could be used as a marker for the unidentified Ags and exploited for molecular monitoring of GVHD posttransplant. To identify alloreactive T cell clones, we performed in vitro allostimulation cultures for a cohort of patients undergoing hemopoietic stem cell transplantation and determined the sequence of the CDR3 of immunodominant alloreactive clones; 10 corresponding clonotypes restricted to activated T cells were identified. As an alternative method for the identification of alloreactive clones, molecular TCR analysis was applied to biopsies of GVHD-affected tissues. Culture- and biopsy-derived clonotypes were used to design sequence-specific quantitative PCR assays to monitor the levels of putative allospecific clonotypes in posttransplant blood samples and subsequent biopsies. Because of the rational design of the methods used to identify immunodominant clonotypes, we were able to follow the behavior of potentially GVHD-specific T cells during the transplant course. Based on our results, we conclude that molecular T cell diagnostics can be a powerful tool for monitoring immune responses posttransplantation.