HLA-A＊0201／Kb及MAGE-3基因共转染B16细胞的制备及其在MAGE-3肿瘤疫苗评价中的作用  

Establishment of B16-HLA-MAGE-3 melanoma cell line coexpressing HLA-A*0201/K^b and MAGE-3 and its role in tumor vaccine evaluation

作　　者：宋淑霞[1] 闫彩珍[2] 郑龙[1] 尤红煜[1] 王俊霞[1] 刘福英[1] 

机构地区：[1]河北医科大学实验动物学部,河北石家庄050017 [2]河北医科大学药理学教研室,河北石家庄050017

出　　处：《细胞与分子免疫学杂志》2008年第7期679-682,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：河北省卫生厅资助项目(04107);河北省科技攻关资助项目(06276102D-30,052761990)

摘　　要：目的:为了用HLA-A2·1转基因鼠制备荷瘤模型以评价肿瘤疫苗的免疫效果,制备共表达HLA-A*0201/Kb嵌合基因和MAGE-3的B16黑色素瘤细胞。方法:采用基因共转染的方法,将HLA-A*0201/Kb和MAGE-3转染到B16黑色素瘤细胞(B16-HLA-MAGE-3),利用RT-PCR、流式细胞术(FCM)和Westernblot检测了HLA-A*0201/Kb和MAGE-3在B16黑色素瘤中的表达;通过测定CTL活性和体内抑瘤实验证实肿瘤抗原MAGE-3可以在B16-HLA-MAGE-3中得到有效加工处理和提呈。结果:RT-PCR、FCM和Westernblot检测到HLA-A*0201/Kb和MAGE-3在B16-HLA-MAGE-3黑色素瘤中的表达;LDH的方法观察到MAGE-3疫苗免疫小鼠脾细胞对B16-HLA-MAGE-3细胞的特异性CTL反应,抑瘤实验也证实,B16-HLA-MAGE-3细胞在免疫小鼠体内生长明显受到抑制。结论:MAGE-3基因在B16-HLA-MAGE-3黑色素瘤细胞中得到表达,并被有效的加工处理、提呈给特异性CD8+T细胞;B16-HLA-MAGE-3细胞可以用来制备荷瘤模型,且该方法适合利用转基因小鼠用于人表位疫苗的体内评价。AIM： To establish a tumor model in HLA- A2.1 transgenic mice to examine the efficacy of MAGE-3 vaccine, a cell line coexpressing HLA-A ＊ 0201/Kb and MAGE-3 is established. METHODS： B16-HLA-MAGE-3 melanoma was obtained by means of cotransfection of HLA- A ＊ 0201/Kb and MAGE-3 to B16 melanoma. RT-PCR, FCM analysis and Western blot were used to detect the mRNA or protein of HLA-A＊ 0201/Kb or MAGE-3 expression in B16- HLA-MAGE-3. The ability of MAGE-3 antigen to be processed and presented in the Bt6-HLA-MAGE-3 cell line were observed by CTL activity detection and tumor chal- lenge test. RESULTS： Transcription and protein expression of HLA-A ＊0201/H-2kb and MAGE-3 were demonstrated in Bt6-HLA-MAGE-3 cells. CTL activity of splenocytes in immunized mice against B16-HLA-MAGE-3 was detected and the growth of B16-HLA-MAGE-3 in immunized mice was also inhibited. CONCLUSION： MAGE-3 antigen is able to be pro- cessed and presented efficiently by B16-HLA-MAGE-3 melanoma cells and this cell can be employed to test HLA-A2 restricted epitope immunogenicity in the A2-transgenic mice.

关 键 词：HLA-A2.1转基因鼠 疫苗评价 肿瘤动物模型 基因共转染 MAGE-3 PIRES 

分 类 号：R392.11[医药卫生—免疫学]