目的:通过双重免疫荧光染色法鉴定不同肺组织内是否存在Clara细胞特异性抗原(Clara cell specific antigen,CCA)和肺泡表面活性蛋白C(surfactant protein C,SP-C)共表达的双阳性细胞(double positive cell,DPCs)CCA+SP-C+细胞,为后续开展肺干细胞的相关研究提供实验基础。方法:取成年鼠(小鼠、大鼠)和新生鼠(小鼠、大鼠)的正常肺组织,人的肺鳞癌、腺癌的癌组织和癌旁组织,冰冻切片,进行双重免疫荧光染色,通过激光扫描共聚焦显微镜观察肺组织内是否存在DPCs。每例标本观察时镜下随机选择4~6个视野,计数100~200个肺细胞,得出每百个细胞内DPCs百分率,采用SPSS 11.0统计软件包对数据进行分析。结果:在成年鼠和新生鼠的正常肺组织中均发现了DPCs的存在,在人的肺腺癌组织中也发现了DPCs,而在鳞癌组织中未见DPCs;在人的癌旁组织DPCs数量明显多于相对应的癌组织(P<0.05)。结论:本研究不仅证实了DPCs细胞存在的广泛性,而且在人的肺腺癌组织中发现了DPCs,同时发现癌旁组织内的DPCs明显多于癌组织,为后续进一步分离、纯化DPCs,研究正常肺干细胞和肺癌干细胞的生物学特性提供了实验基础。
Objective： With the development of peptide-based cancer specific immunotherapy, the prediction of CTL epitopes from insulin-like growth factor-binding protein 7 （IGFBP7） is very important for some research about tumor metastasis. Because HLA-A2.1-expressing individuals cover 〉50% in the population of China, we aimed at identifying IGFBPT-encoded peptide presented by HLA-A2.1. Methods： In our study, a HLA-A2.1 restricted CTL epitope was identified by using the following two-step procedure： （a） computer-based epitope prediction from the amino acid sequence of IGFBP7 antigen; （b） Validation with epitope molecular modeling. Results： We obtained four epitopes with high immunogenicity scores by all of the three algorithms, i.e., BIMAS, SYFPEITH1 and IMTECH. Each of the four candidates satisfied the criteria of the HLA-A2.1- restricted CTL epitopes in molecular modeling analysis. Conclusion： The combination of BIMAS, SYFPEITHI and IMTECH method can improve the prediction efficiency and accuracy. Due to this research herein, this four epitopes have potential value for further studied, also have potential application in peptide-mediated immunotherapy. These epitopes may be useful in the design of therapeutic peptide vaccine for lung carcinoma and as immunotherapeutic strategies against lung carcinoma after identified by immunology experiment.
