Storkus W J, Zeh H J, Maeurer M J, Salter R D, Lotze M T
Department of Surgery, University of Pittsburgh School of Medicine, PA 15261.
J Immunol. 1993 Oct 1;151(7):3719-27.
Using a newly described pH 3.3 acid elution technique, peptides were extracted by denaturation of class I molecules on the surface of human melanomas. HPLC fractionation of this material revealed six T cell epitopes (termed P1-P6) recognized by HLA-A2-restricted, melanoma-specific tumor infiltrating lymphocyte (TIL) lines. Three of these fractions (P1, P2, and P4) appeared to represent shared/immunodominant melanoma Ag recognized in the context of HLA-A2 because they were expressed by 4/4 HLA-A2+ melanoma cell lines and were each recognized by all four oligoclonal HLA-A2-restricted TIL lines examined. Interestingly, P1 and P2 (but not P3-P6) could also be recognized by these same TIL when presented by the HLA-Aw69 class I molecule, which is closely related to HLA-A2. P3, P5, and P6 displayed more restricted expression and were differentially recognized by the four oligoclonal TIL lines. These results suggest that synthetic peptide derived from P1, P2, and P4 sequences (when deduced) may form the basis of effective prophylactic or therapeutic melanoma vaccines by stimulating CD8+ CTL in HLA-A2+ individuals. This approach of identifying T cell epitopes presented by class I molecules should prove generally applicable to the study of other tumors recognized by class I-restricted CTL.
采用一种新描述的pH 3.3酸洗脱技术，通过对人黑色素瘤表面的I类分子进行变性来提取肽段。对该材料进行高效液相色谱分级分离，揭示了6个T细胞表位（称为P1 - P6），它们被HLA - A2限制性、黑色素瘤特异性肿瘤浸润淋巴细胞（TIL）系所识别。其中三个组分（P1、P2和P4）似乎代表了在HLA - A2背景下被识别的共享/免疫显性黑色素瘤抗原，因为它们在4/4的HLA - A2 +黑色素瘤细胞系中表达，并且在所有检测的四个寡克隆HLA - A2限制性TIL系中均被识别。有趣的是，当由与HLA - A2密切相关的HLA - Aw69 I类分子呈递时，这些相同的TIL也能识别P1和P2（但不是P3 - P6）。P3、P5和P6表现出更受限的表达，并且被四个寡克隆TIL系差异识别。这些结果表明，源自P1、P2和P4序列（推导时）的合成肽可能通过刺激HLA - A2 +个体中的CD8 + CTL，构成有效的预防性或治疗性黑色素瘤疫苗的基础。这种识别I类分子呈递的T细胞表位的方法应该证明普遍适用于研究其他被I类限制性CTL识别的肿瘤。
