University Cell Immunotherapy Center, Masaryk University, Brno, Czech Republic.
Med Sci Monit. 2009 Dec;15(12):CR638-43.
Renal cell carcinomas have developed various strategies to escape immune cell recognition, including down-regulation or loss of classic HLA class I antigens (A, B, C) and aberrant expression of non-classic HLA class I antigens (G, E). In this study both classic and non-classic HLA class I antigens were tested in tumor specimens and established primary cell cultures derived from renal cell carcinoma patients.
MATERIAL/METHODS: HLA class I antigens were evaluated by immunohistochemical staining and the intensity of cytoplasmic staining was measured semiquantitatively. Renal tumor tissue obtained from nephrectomy was used for the explant culture. MTT assay was performed to test the chemoresistance of primary cell line cultures to common cytostatics.
HLA-G and HLA-E were found in 62% and 100% of the analyzed tumor samples, respectively. Markedly higher levels of the non-classic HLA-G and -E antigens compared with the classic HLA-A, -B, and -C antigens were observed. The cells of the control renal tissues were HLA-A, -B, -C, and -E positive and HLA-G negative. Cell line cultures were successfully established in 85% of the renal cell carcinoma specimens. No or minimal changes in classic HLA-A, B, and C antigen staining were observed during cultivation of the primary cell line cultures. No correlation between HLA class I antigen expression and chemoresistance, histopathological stage, or nuclear grade was found.
These findings suggest that primary cell line cultures derived from surgical specimens of renal cell carcinomas are a feasible model for immunotherapy research through their high cultivation potential.
肾细胞癌已开发出各种策略来逃避免疫细胞的识别，包括下调或丧失经典 HLA Ⅰ类抗原（A、B、C）和异常表达非经典 HLA Ⅰ类抗原（G、E）。在这项研究中，对肿瘤标本和源自肾细胞癌患者的建立的原代细胞培养物进行了经典和非经典 HLA Ⅰ类抗原检测。
材料/方法：通过免疫组织化学染色评估 HLA Ⅰ类抗原，并用半定量方法测量细胞质染色的强度。从肾切除术获得的肾肿瘤组织用于外植体培养。MTT 测定用于测试原代细胞系培养物对常见细胞毒药物的化学抗性。
在分析的肿瘤样本中，分别发现 HLA-G 和 HLA-E 为 62%和 100%。与经典 HLA-A、B 和 C 抗原相比，观察到非经典 HLA-G 和 -E 抗原的水平明显更高。对照肾组织的细胞均为 HLA-A、B、C 和 -E 阳性，HLA-G 阴性。在 85%的肾细胞癌标本中成功建立了细胞系培养物。在原代细胞系培养过程中，经典 HLA-A、B 和 C 抗原染色没有或仅有微小变化。未发现 HLA Ⅰ类抗原表达与化学抗性、组织病理学分期或核级之间存在相关性。
这些发现表明，源自肾细胞癌手术标本的原代细胞系培养物通过其高培养潜力成为免疫治疗研究的可行模型。
