Saikali Stéphan, Avril Tony, Collet Brigitte, Hamlat Abderrahmane, Bansard Jean-Yves, Drenou Bernard, Guegan Yvon, Quillien Véronique
Département d'Anatomie et cytologie pathologiques, Hôpital Pontchaillou, Rennes, France.
J Neurooncol. 2007 Jan;81(2):139-48. doi: 10.1007/s11060-006-9220-3. Epub 2006 Sep 27.
In this study, we investigated the mRNA and protein expression of nine tumour antigens in human glioblastoma multiforme with a view to their possible use in dendritic cell-based immunotherapy. Expression of ALK, EGFRvIII, GALT3, gp100, IL-13Ralpha2, MAGE-A3, NA17-A, TRP-2 and tyrosinase were studied by real-time RT-PCR on frozen tissues using a series of 47 tumour samples from patients with glioblastoma. Results were compared with non-neoplastic brain expression or glioblastoma samples with very low levels of expression near the limits of detection for EGFRvIII and MAGE-A3, as these latter two antigens were not detected in non-neoplastic brain. Tumour antigens showing a 5-fold increase in mRNA expression were considered as positive, and only antigens displaying an mRNA over-expression in a significant number of cases were analysed by immunohistochemistry on paraffin-embedded sections. Using real time RT-PCR, we found EGFRvIII, gp100, IL-13Ralpha2 and TRP-2 to be positive in 64, 38, 32 and 21% of cases, respectively. While we observed no over-expression for ALK, GALT3 and tyrosinase, 3 samples out of 47 were positive for MAGE-3 and 1 sample for NA17-A. More than 25% of tumour cells showed strong protein expression in 13, 34, 85 and 96% of GBM samples for gp100, TRP-2, EGFRvIII and IL-13Ralpha2, respectively. Interestingly, protein expression of at least 3 antigens was observed in 38% of cases. These results point out the importance of EGFRvIII, IL-13Ralpha2 and, to a less extent gp100 and TRP-2, for developing an immunotherapy strategy against glioblastoma.
在本研究中，我们调查了9种肿瘤抗原在多形性胶质母细胞瘤中的mRNA和蛋白表达情况，以期其能用于基于树突状细胞的免疫治疗。使用来自47例胶质母细胞瘤患者的冷冻组织，通过实时逆转录聚合酶链反应（RT-PCR）研究了间变性淋巴瘤激酶（ALK）、表皮生长因子受体III型变异体（EGFRvIII）、半乳糖凝集素3（GALT3）、糖蛋白100（gp100）、白细胞介素13受体α2（IL-13Rα2）、黑色素瘤相关抗原A3（MAGE-A3）、NA17-A、酪氨酸酶相关蛋白2（TRP-2）和酪氨酸酶的表达。将结果与非肿瘤性脑组织的表达情况或EGFRvIII和MAGE-A3表达水平极低接近检测限的胶质母细胞瘤样本进行比较，因为在非肿瘤性脑组织中未检测到后两种抗原。mRNA表达增加5倍的肿瘤抗原被视为阳性，仅对在大量病例中显示mRNA过表达的抗原进行石蜡包埋切片的免疫组织化学分析。通过实时RT-PCR，我们发现EGFRvIII、gp100、IL-13Rα2和TRP-2分别在64%、38%、32%和21%的病例中呈阳性。虽然我们未观察到ALK、GALT3和酪氨酸酶的过表达，但47个样本中有3个MAGE-3呈阳性，1个NA17-A呈阳性。在胶质母细胞瘤样本中，分别有13%、34%、85%和96%的样本中超过25%的肿瘤细胞显示gp100、TRP-2、EGFRvIII和IL-13Rα2有强蛋白表达。有趣的是，38%的病例中观察到至少3种抗原的蛋白表达。这些结果指出了EGFRvIII、IL-13Rα2以及在较小程度上gp100和TRP-2对于制定抗胶质母细胞瘤免疫治疗策略的重要性。
