Mosolits Szilvia, Markovic Katja, Fagerberg Jan, Frödin Jan-Erik, Rezvany Mohammad-Reza, Kiaii Shahryar, Mellstedt Håkan, Jeddi-Tehrani Mahmood
Immune and Gene Therapy Laboratory, Department of Oncology (Radiumhemmet), Karolinska Institute, Stockholm, Sweden.
Cancer Immunol Immunother. 2005 Jun;54(6):557-70. doi: 10.1007/s00262-004-0620-y. Epub 2004 Nov 27.
The tumour-associated antigen, Ep-CAM, is over-expressed in colorectal carcinoma (CRC). In the present study, a recombinant Ep-CAM protein or a human anti-idiotypic antibody (anti-Id) mimicking Ep-CAM, either alone or in combination, was used for vaccination of CRC patients (n=9). GM-CSF was given as an adjuvant cytokine. A cellular immune response was assessed by measuring anti-Ep-CAM lymphoproliferation, IFN-gamma production (ELISPOT) and by analysing the TCR BV gene usage within the CD4+ and CD8+ T-cell subsets followed by CDR3 fragment analysis. A proliferative and/or IFN-gamma T-cell response was induced against the Ep-CAM protein in eight out of nine patients, and against Ep-CAM-derived peptides in nine out of nine patients. Analysis of the TCR BV gene usage showed a significantly higher usage of BV12 family in CD4+ T cells of patients both before and after immunisation than in those of healthy control donors (p<0.05). In the CD8+ T-cell subset, a significant (p<0.05) increase in the BV19 usage was noted in patients after immunisation. In individual patients, a number of TCR BV gene families in both CD4+ and CD8+ T cells were over-expressed mainly in post-immunisation samples. Analysis of the CDR3 length polymorphism revealed a higher degree of clonality in post-immunisation samples than in pre-immunisation samples. In vitro stimulation with Ep-CAM protein confirmed the expansion of anti-Ep-CAM T-cell clones. The results indicate that immunisation with the Ep-CAM protein and/or anti-Id entails the induction of an anti-Ep-CAM T-cell response in CRC patients, and suggest that BV19+ CD8+ T cells might be involved in a vaccine-induced immune response.
肿瘤相关抗原Ep-CAM在结直肠癌（CRC）中过度表达。在本研究中，重组Ep-CAM蛋白或模拟Ep-CAM的人抗独特型抗体（抗Id）单独或联合用于CRC患者（n = 9）的疫苗接种。给予GM-CSF作为佐剂细胞因子。通过测量抗Ep-CAM淋巴细胞增殖、IFN-γ产生（ELISPOT）以及分析CD4 +和CD8 + T细胞亚群内的TCR BV基因使用情况，随后进行CDR3片段分析来评估细胞免疫反应。9名患者中有8名对Ep-CAM蛋白诱导了增殖性和/或IFN-γ T细胞反应，9名患者中有9名对Ep-CAM衍生肽诱导了反应。TCR BV基因使用情况分析显示，免疫前后患者CD4 + T细胞中BV12家族的使用显著高于健康对照供体（p<0.05）。在CD8 + T细胞亚群中，免疫后患者的BV19使用显著增加（p<0.05）。在个体患者中，CD4 +和CD8 + T细胞中的一些TCR BV基因家族主要在免疫后样本中过度表达。CDR3长度多态性分析显示，免疫后样本中的克隆性程度高于免疫前样本。用Ep-CAM蛋白进行体外刺激证实了抗Ep-CAM T细胞克隆的扩增。结果表明，用Ep-CAM蛋白和/或抗Id进行免疫可在CRC患者中诱导抗Ep-CAM T细胞反应，并提示BV19 + CD8 + T细胞可能参与疫苗诱导的免疫反应。
