Evaluation of antigen-specific responses using in vitro enriched T cells
Evaluation of antigen-specific responses using in vitro enriched T cells
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DOI：
10.1016/s0022-1759(02)00510-0
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发表时间：
2003-03-01
影响因子：
2.2
通讯作者：
Cao, HY
中科院分区：
文献类型：
--
作者：
Jones, N;Agrawal, D;Cao, HY
抗原特异性淋巴细胞在对病毒感染的免疫应答中是重要的。在大多数免疫学研究中，外周血单核细胞（PBMC）传统上用作效应细胞的来源。本文描述了使用双特异性单克隆抗体（BSMAB）抗CD3：CD8（CD3，8）和抗CD3：CD4（CD3，413）分别扩增和选择性富集CD4+和CD8+ T细胞群。扩增的细胞在14天时显示>90%的⑶ 3 + ⑶ 4+或⑶ 3 + ⑶ 8+。我们测量了这些亚群富集的T细胞的HIV和CMV特异性应答，发现在各种细胞免疫学测定（CMI）中与PBMC相比，灵敏度和特异性相似或更高。BSMAB富集的细胞的Vbeta分析表明与亲本PBMC相当的库。尽管用BSMAB扩增了CD45RA（hi）和CD45RO（hi）细胞群，但选择性亚群耗竭表明抗原特异性T细胞应答仅限于初始CD45RO（hi）记忆效应子亚群。总之，BSMAB体外富集T细胞允许显著扩增细胞群而不丧失特异性。这种细胞扩增技术允许在初始细胞来源稀缺的情况下研究T细胞亚群功能，并为免疫测定中的活的和功能性T细胞提供了替代方案。(C)2002 Elsevier Science B.V.保留所有权利。
Antigen-specific lymphocytes are important in the immune response to viral infection. Peripheral blood mononuclear cells (PBMC) are traditionally used as a source of effector cells in most immunological studies. We described here the use of the bispecific monoclonal antibodies (BSMAB) anti CD3:CD8 (CD3,8) and anti CD3:CD4 (CD3,413) to expand and selectively enrich CD4+ and CD8+ T cell populations, respectively. The expanded cells demonstrated >90% CD3+ CD4+ or CD3+ CD8+ by 14 days. We measured HIV- and CMV-specific responses of these subset-enriched T cell and found that sensitivity and specificity is similar or higher when compared to PBMC in various cellular immunology assays (CMI). Vbeta analysis of BSMAB-enriched cells demonstrated comparable repertoir to the parent PBMC. Although both CD45RA(hi) and CD45RO(hi) cell populations were expanded with the BSMAB, selective subset depletion demonstrated that the antigen-specific T cell responses were restricted to the initial CD45RO(hi) memory effector subgroup. In conclusion, BSMAB in vitro enrichment of T cells allows significant expansion of the cell population without loss of specificity. This technique of cell expansion permits studies of T cell subset function in situations where the initial cell source is scarce, and presents an alternative for viable and functional T cells in immunological assays. (C) 2002 Elsevier Science B.V. All rights reserved.