Induction of CD4+ Th1 lymphocytes that recognize known and novel class 11 MHC restricted epitopes from the melanoma antigen gp100 by stimulation with recombinant protein
Induction of CD4+ Th1 lymphocytes that recognize known and novel class 11 MHC restricted epitopes from the melanoma antigen gp100 by stimulation with recombinant protein
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DOI：
10.1097/00002371-200403000-00001
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发表时间：
2004-03-01
影响因子：
3.9
通讯作者：
Rosenberg, SA
中科院分区：
文献类型：
--
作者：
Parkhurst, MR;Riley, JP;Rosenberg, SA
CD4(+) T辅助细胞可能在诱导和维持对癌症的治疗性免疫反应中发挥关键作用。为了评估重组肿瘤相关蛋白诱导抗原反应性CD4(+) T细胞的功效，我们在体外用大肠杆菌纯化的黑色素瘤抗原gp100刺激黑色素瘤患者的外周血淋巴细胞。在初步实验中，我们观察到外周血单核细胞在外源加载蛋白后可以加工并将已知的HLA-DRbeta1*0401和HLA-DRbeta1*0701限制性表位呈递到gp100反应性CD4(+) T细胞I系。因此，我们使用自体载蛋白的外周血单个核细胞作为抗原提呈细胞。从9名同时表达HLA-DRbeta1*0401和HLA-DRbeta1*0701的患者中的4名患者中，我们培养了5个gp100反应性CD4(+) T细胞群，这些细胞群分泌T， I型细胞因子以响应外源性负载蛋白，以及内源性表达gp100和MHC 11类分子的靶细胞，包括转染细胞和黑色素瘤细胞。5个培养中有4个特异性识别出已知的HLA-DRbeta1*0401和HLA-DRbeta1*0701限制性表位，分别为gp100:44-59和gp100:170-190。第五次培养，以及由此衍生的30个T细胞克隆，在HLA-DRbeta1*0701的背景下特异性识别新的肽gp100:420-435。这些结果表明，重组肿瘤相关蛋白可能在临床上适用于主动疫苗接种策略或过继细胞免疫疗法中产生CD4(+) T辅助细胞。
CD4(+) T helper cells may play a critical role in the induct on and maintenance of a therapeutic immune response to cancer. To evaluate the efficacy with which a recombinant tumor-associated protein can induce antigen-reactive CD4(+) T cells, we stimulated peripheral blood lymphocytes from patients with melanoma in vitro with the purified melanoma antigen gp100 produced in Escherichia coli. In preliminary experiments, we observed that peripheral blood mononuclear cells could process and present known HLA-DRbeta1*0401 and HLA-DRbeta1*0701 restricted epitopes to gp100-reactive CD4(+) T cell I ties after being loaded exogenously with protein. Therefore, we used autologous protein-loaded peripheral blood mononuclear cells as antigen presenting cells. From four of nine patients who expressed both HLA-DRbeta1*0401 and HLA-DRbeta1*0701, we raised five gp100-reactive CD4(+) T cell populations that secreted T, I type cytokines in response to exogenously loaded protein as well as target cells that endogenously expressed gp100 and MHC class 11 molecules, including transfectants and melanoma cells. Four of the five cultures specifically recognized the known HLA-DRbeta1*0401 and HLA-DRbeta1*0701 restricted epitopes gp100:44-59 and gp100:170-190, respectively. The fifth culture, and 30 T cell clones derived from it, specifically recognized anew peptide, gp100:420-435, in the context of HLA-DRbeta1*0701. These results Suggest that recombinant tumor-associated proteins may be clinically applicable for the generation of CD4(+) T helper cells in active vaccination strategies or adoptive cellular immunotherapies.