Combination of MHC-peptide multimer-based T cell sorting with the immunoscope permits sensitive ex vivo quantitation and follow-up of human CD8+ T cell immune responses
Combination of MHC-peptide multimer-based T cell sorting with the immunoscope permits sensitive ex vivo quantitation and follow-up of human CD8+ T cell immune responses
复制标题
DOI：
10.1016/s0022-1759(02)00004-2
复制
发表时间：
2002-03-01
影响因子：
2.2
通讯作者：
Pannetier, C
中科院分区：
文献类型：
--
作者：
Lim, A;Baron, V;Pannetier, C
MHC限制性抗原的鉴定以及适应性细胞毒性免疫应答的诱导和控制的进展已经引起了对免疫疗法作为严重病理如癌症和自身免疫性疾病的治疗的新兴趣。需要在整个临床试验中检测和监测由治疗诱导的T细胞应答的可靠程序，以便设计具有更高效率的合理方案。我们已经尝试开发这样一个程序相结合的T细胞分选使用HLA-肽复合物多聚化的磁珠连同定量免疫方法。一旦招募的患者已经针对HLA和靶抗原进行了分型，就可以选择相关的HLA-肽多聚体并用于在任何治疗之前和在对治疗的预期应答的峰值时分选特定的外周T细胞。然后可以设计对特定T细胞克隆的TCR重排具有特异性的克隆型引物，并用于在整个试验中通过对血液样品或T细胞亚群进行定量PCR来测量其TCR转录物的频率。在重建实验中以及在一个类风湿性关节炎患者的样本中，我们能够很容易地检测和跟踪几个T细胞克隆，其频率在CD 8（+）T细胞中低至10（-5）。该方法相对于其他现有检测方法的主要优点是，它不需要对特定T细胞的功能状态进行任何假设，并且它允许监测单个T细胞克隆，从而可以评估其表型转变。(C)2002 Elsevier Science B. V.保留所有权利。
Identification of MHC-restricted antigens and progress in the induction and control of adaptive cytotoxic immune responses have led to renewed interest in immunotherapy as a treatment for severe pathologies such as cancer and autoimmune diseases. Reliable procedures for detecting and monitoring T cell responses induced by the treatment throughout a clinical trial are needed in order to design rational protocols with increased efficiency. We have attempted to develop such a procedure by combining T cell sorting using HLA-peptide complexes multimerized on magnetic beads together with the quantitative Immunoscope approach. Once a recruited patient has been typed for HLA and target antigens, relevant HLA-peptide multimers can be selected and used for sorting specific peripheral T cells prior to any treatment and at the peak of the expected response to treatment. Clonotypic primers specific for the TCR rearrangements of the specific T cell clones can then be designed and used for measuring the frequency of their TCR transcripts by quantitative PCR on blood samples or T cell subsets throughout the trial. In reconstruction experiments as well as in samples from one rheumatoid arthritis patient, we were readily able to detect and follow several T cell clones with a frequency as low as 10(-5) among CD8(+) T cells. The main advantages of this procedure over other currently available assays are that it does not require any assumptions on the functional status of the specific T cells and it permits the monitoring of individual T cell clones whose phenotypic shift can thus be evaluated. (C) 2002 Elsevier Science B.V. All rights reserved.