Schmitt Anita, Barth Thomas F E, Beyer Eva, Borchert Franziska, Rojewski Markus, Chen Jinfei, Guillaume Philippe, Gronau Silke, Greiner Jochen, Möller Peter, Riechelmann Herbert, Schmitt Michael
Department of Internal Medicine III, University of Ulm, Ulm, Germany.
Int J Oncol. 2009 Mar;34(3):629-39. doi: 10.3892/ijo_00000188.
Despite advances in surgery, radio- and chemotherapy, therapeutic approaches for patients with head and neck squamous carcinoma (HNSCC) need to be improved. Immunotherapies eliciting tumor specific immune responses might constitute novel treatment options. We therefore investigated the expression and immunogenicity of two tumor-associated antigens (TAA) the receptor for hyaluronic acid mediated motility (RHAMM) and carboanhydrase IX (G250/CAIX) in HNSCC patients. Twenty-two HNSCC samples were examined for the expression of RHAMM and G250 by Western blotting and immunohistochemistry, 14/22 samples were tested for HLA-A2 expression by flow cytometry. For 8/22 samples single tumor-cell suspensions were generated, and mixed lymphocyte peptide cultures (MLPC) were performed to evaluate the frequencies of cytotoxic T cells specifically recognizing RHAMM and G250 using Tetramer staining/multi-color flow cytometry and enzyme linked immunosorbent spot (ELISPOT) assays. RHAMM and G250 were expressed in 73 and 80% of the HNSCC samples at the protein level. A co-expression of both TAAs could be detected in 60% of the patients. In 4/8 HLA-A2+ patients, 0.06-0.13% of CD8+ effector T cells recognized Tetramers for RHAMM or G250 and secreted IFNgamma and granzyme B in ELISPOT assays. RHAMM and G250 are expressed at high frequency and high protein level in HNSCCs and are recognized by cytotoxic CD8+ effector T cells. Therefore both TAAs constitute interesting targets for T cell based immunotherapies for HNSCC.
尽管在手术、放疗和化疗方面取得了进展，但头颈部鳞状细胞癌（HNSCC）患者的治疗方法仍需改进。引发肿瘤特异性免疫反应的免疫疗法可能构成新的治疗选择。因此，我们研究了两种肿瘤相关抗原（TAA）——透明质酸介导的运动受体（RHAMM）和碳酸酐酶IX（G250/CAIX）在HNSCC患者中的表达和免疫原性。通过蛋白质印迹法和免疫组织化学检查了22例HNSCC样本中RHAMM和G250的表达，通过流式细胞术检测了14/22例样本中的HLA-A2表达。对于8/22例样本，制备了单个肿瘤细胞悬液，并进行了混合淋巴细胞肽培养（MLPC），以使用四聚体染色/多色流式细胞术和酶联免疫吸附斑点（ELISPOT）测定法评估特异性识别RHAMM和G250的细胞毒性T细胞的频率。RHAMM和G250在73%和80%的HNSCC样本中呈蛋白质水平表达。在60%的患者中可检测到两种TAA的共表达。在4/8例HLA-A2+患者中，0.06-0.13%的CD8+效应T细胞在ELISPOT测定中识别RHAMM或G250的四聚体并分泌IFNγ和颗粒酶B。RHAMM和G250在HNSCC中高频且高蛋白水平表达，并被细胞毒性CD8+效应T细胞识别。因此，这两种TAA均构成基于T细胞的HNSCC免疫疗法的有趣靶点。
