Takaki T, Hiraki A, Uenaka A, Gomi S, Itoh K, Udono H, Shibuya A, Tsuji T, Sekiguchi S, Nakayama E
Department of Internal Medicine, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700, Japan.
Int J Oncol. 1998 May;12(5):1103-9. doi: 10.3892/ijo.12.5.1103.
Cytotoxic T lymphocytes (CTL) specific for HLA-A2-binding MAGE-3 peptide (FLWGPRALV) were generated by repetitive stimulation of PBMC with the peptide in the presence of EBV-transformed B blasts and IL-2. Using these CTL, we investigated the expression of the HLA-A2-binding MAGE-3 peptide on lung cancer cell lines. Of 14 cell lines investigated, 1-87, PC-9, OU-LC-KI, 11-18 and LK87 were derived from HLA-A2 positive patients. But cytofluorometry analysis showed that 1-87, PC-9 and OU-LC-KI, but not 11-18 or LK87 expressed the HLA-A2 antigen. All five cell lines expressed MAGE-3 gene mRNA. Twelve of thirteen CTL lines from two HLA-A2 positive donors showed no cytotoxicity against any of the 14 lung cancer cell lines. CTL line TI-1 showed cytotoxicity against 1-87 but not against any of the other cell lines. Treatment of 1-87 with IFN-gamma greatly augmented the cytotoxicity of TI-1 and induced it in the other 12 CTL lines, confirming the expression of the peptide on 1-87. No cytotoxicity was induced by IFN-gamma treatment of PC-9 or OU-LC-KI. However, PC-9 and OU-LC-KI pulsed with the peptide were killed efficiently by all of the CTL lines, suggesting no expression of the peptide on those cells. A low level of cytotoxicity was induced on 11-18 but not LK87 by IFN-gamma treatment, although expression of the HLA-A2 antigen was not observed by cytofluorometry. These findings showed that expression of the HLA-A2-binding MAGE-3 peptide recognized by CTL was variable on lung cancer cell lines.
通过在EB病毒转化的B淋巴细胞和白细胞介素-2存在的情况下，用HLA - A2结合的MAGE - 3肽（FLWGPRALV）反复刺激外周血单核细胞（PBMC），产生了对该肽具有特异性的细胞毒性T淋巴细胞（CTL）。利用这些CTL，我们研究了HLA - A2结合的MAGE - 3肽在肺癌细胞系上的表达情况。在所研究的14个细胞系中，1 - 87、PC - 9、OU - LC - KI、11 - 18和LK87来源于HLA - A2阳性患者。但细胞荧光分析显示，1 - 87、PC - 9和OU - LC - KI表达HLA - A2抗原，而11 - 18和LK87不表达。所有这五个细胞系均表达MAGE - 3基因mRNA。来自两名HLA - A2阳性供体的13个CTL系中的12个对14个肺癌细胞系中的任何一个均无细胞毒性。CTL系TI - 1对1 - 87具有细胞毒性，但对其他任何细胞系均无细胞毒性。用γ干扰素处理1 - 晚期宫颈癌的治疗方案87可大大增强TI - 1的细胞毒性，并在其他12个CTL系中诱导产生细胞毒性，证实了该肽在1 - 87上的表达。用γ干扰素处理PC - 9或OU - LC - KI未诱导出细胞毒性。然而，用该肽脉冲处理的PC - 9和OU - LC - KI被所有CTL系有效杀伤，表明这些细胞上不表达该肽。用γ干扰素处理后，11 - 18诱导出低水平的细胞毒性，但LK87未被诱导，尽管通过细胞荧光分析未观察到HLA - A2抗原的表达。这些发现表明，CTL识别的HLA - A2结合的MAGE - 3肽在肺癌细胞系上的表达是可变的。
