In vitro generation of human cytotoxic T lymphocytes specific for peptides derived from prostate-specific antigen
In vitro generation of human cytotoxic T lymphocytes specific for peptides derived from prostate-specific antigen
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DOI：
10.1093/jnci/89.4.293
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发表时间：
1997-02-19
影响因子：
10.3
通讯作者：
Tsang, KY
中科院分区：
文献类型：
--
作者：
Correale, P;Walmsley, K;Tsang, KY
工作背景：蛋白质抗原作为与主要组织相容性复合物的I类分子结合的小肽（约9-10个氨基酸长）呈递给细胞毒性T淋巴细胞。抗原决定簇）可用于抗癌疫苗的开发。对一个肽表位的细胞毒性T细胞应答的产生目的：我们的目的是鉴定能够引发特异性细胞毒性T细胞应答的新型PSA肽。方法：基于以下标准鉴定候选肽：1）它们含有用于结合HLA-A2的共有氨基酸基序，HLA-A2是最常见类型的I类分子; 2）它们与PSA相关的激肽释放酶蛋白缺乏强同源性;和3）它们能够稳定人T2（转运缺失突变体）细胞表面上的HLA-A2 I类分子，所述细胞在抗原呈递方面有缺陷，能够杀伤（即，裂解）通过将外周血单核细胞与肽和白细胞介素2一起孵育，从三个不同的男性（两个无疾病个体和一个患有前列腺癌的患者）产生已经用特异性PSA肽脉冲的T2靶细胞。通过从已经用[In-111]羟基喹啉标记的靶细胞释放放射性来监测特异性细胞裂解。结果如下：鉴定了两种能够引发细胞毒性T细胞应答的新型PSA肽;这些肽被命名为PSA-1（氨基酸141-150）和PSA-3（氨基酸154-163），响应于这些肽产生了四种不同的细胞毒性T细胞系-三种针对PSA-3，一种针对PSA-1，通过添加针对I类分子的单克隆抗体，T细胞系对肽脉冲的T2细胞的特异性裂解被阻断。T细胞系还能够裂解PSA阳性、HLA-Aa阳性LNCaP细胞LNCaP细胞裂解的特异性由以下显示：1）添加的人K562不能（慢性骨髓性白血病）细胞抑制它的能力，2）添加的抗HLA-A2抗体阻断它的能力，和3）T细胞系不能诱导PSA阴性的大量溶解，HLA-A2阳性的人类癌细胞，意义：我们的研究形成了一个合理的基础上使用PSA肽或重组载体编码PSA的抗癌疫苗免疫治疗方案的发展与前列腺癌患者。
Background: Protein antigens are presented to cytotoxic T lymphocytes as small peptides (approximately 9-10 amino acids long) bound to class I molecules of the major histocompatibility complex, The identification of tumor-associated antigens and specific peptide epitopes (i.e., antigenic determinants) may be useful in the development of anticancer vaccines, The generation of a cytotoxic T-cell response to one peptide epitope (amino acids 146-154) of human prostate-specific antigen (PSA) has been reported, Purpose: Our aim was to identify novel PSA peptides capable of eliciting specific cytotoxic T-cell responses, Methods: Candidate peptides were identified on the basis of the following criteria: 1) they contained consensus amino acid motifs for binding to HLA-A2, which is the most common type of class I molecule; 2) they lacked strong homology with PSA-related kallikrein proteins; and 3) they were capable of stabilizing HLA-A2 class I molecules on the surface of human T2 (transport deletion mutant) cells, which are defective in antigen presentation, T-cell lines capable of killing (i.e., lysing) T2 target cells that had been pulsed with specific PSA peptides were generated from three different males (two disease-free individuals and one patient with prostate cancer) by incubating peripheral blood mononuclear cells with the peptides and interleukin 2, Specific cell lysis was monitored by the release of radioactivity from target cells that had been labeled with [In-111]oxyquinoline. Results: Two novel PSA peptides capable of eliciting cytotoxic T-cell responses were identified; these peptides were designated PSA-1 (amino acids 141-150) and PSA-3 (amino acids 154-163), Four different cytotoxic T-cell lines were generated in response to these peptides-three against PSA-3 and one against PSA-1, Specific lysis of peptide-pulsed T2 cells by the T-cell lines was blocked by the addition of a monoclonal antibody directed against class I molecules, The T-cell lines were also capable of lysing PSA-positive, HLA-Aa-positive LNCaP cells (human prostate carcinoma cells), The specificity of LNCaP cell lysis was shown by the following: 1) the inability of added human K562 (chronic myelogenous leukemia) cells to inhibit it, 2) the ability of added anti-HLA-A2 antibodies to block it, and 3) the inability of the T-cell lines to induce substantial lysis of PSA-negative, HLA-A2-positive human cancer cells, Implications: Our studies form a rational basis for the use of PSA peptides or recombinant vectors encoding PSA in the development of anticancer vaccine immunotherapy protocols for patients with prostate cancer.