Basic study of tumor-vaccine for metastatic epithelial cancer.

利用ke4细胞毒性T细胞（CTL）系，通过基因表达克隆方法鉴定了一个lck基因。KE4-CTL系先前从鳞状细胞癌的肿瘤浸润淋巴细胞（til）中建立。Lck蛋白（p56k/c）是一种src家族酪氨酸激酶，对T细胞的发育和功能至关重要，在转移性结肠癌中异常表达。I型转录本用于癌细胞系，与先前报道的结果一致，而II型转录本用于正常细胞和组织。此外，所有的p56^<lck>食管癌细胞系都是从有不同转移的原发性食管癌患者中建立的，包括克隆了lck基因作为编码肿瘤排斥抗原基因的KE4肿瘤细胞。P56 ^<lck>似乎通过启动非锚定增殖促进上皮细胞的恶性转化。我们鉴定了3个肽段（Lck208-216， Lck486-494和Lck488-497），它们被KE4-CTL识别。这些Lck肽增强了结肠癌和其他有转移的上皮癌患者外周血单核细胞中CTh的活性，而没有转移的患者则没有。识别Lck肽的CTL前体在转移性癌症患者的新鲜制备的pbmc中被发现，并且它们的频率在肽刺激下显着增加。因此，Lck肽可用于转移性癌症患者的特异性免疫治疗。

A lck gene was identified by the gene-expression cloning method, in which a KE4-cytotoxic T cell (CTL) line was used. The KE4-CTL line was previously established from tumor-infiltrating lymphocytes (TILs) of squamous cell carcinoma. The Lck protein (p56k/c), a src family tyrosine kinase which is essential for T cell development and function, is aberrantly expressed in metastatic colon cancers. The type I transcript was used in the cancer cell lines, in agreement with the results reported previously, whereas the type II transcript was used in normal cells and tissues.Further, all p56^<lck> esophageal cancer cell lines were established from the primary esophageal tumors patients who had distinct metastases, including the KE4 tumor cells from which the lck gene was cloned as a gene encoding tumor-rejection antigen. p56^<lck> seems to facilitate the malignant transformation of epithelial cells through initiation of anchorage-independent proliferation. We identified three peptides (Lck208-216, Lck486-494, and Lck488-497), which were recognized by KE4-CTL. These Lck peptides augmented CTh activity in peripheral blood mononuclear cells of colon and other epithelial cancer patients with metastasis, but not those without metastasis. CTL precursors recognizing the Lck peptide were identified in freshly prepared PBMCs of metastatic cancer patients, and their frequency was significantly augmented by the stimulation with the peptide. Thus, Lck peptides could be useful for specific immunotherapy toward metastatic cancer patients.