[共表达HLA-A 0201/K(b)和MAGE-3的B16-HLA-MAGE-3黑色素瘤细胞系的建立及其在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].

作者信息

Song Shu-Xia, Yan Cai-Zhen, Zheng Long, You Hong-Yu, Wang Jun-Xia, Liu Fu-Ying

机构信息

Department of Animal Laboratory, Hebei Medical University, Shijiazhuang 050017, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jul;24(7):679-82.

PMID:18616910

Abstract

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/K(b) and MAGE-3 is established.

METHODS

B16-HLA-MAGE-3 melanoma was obtained by means of cotransfection of HLA-A 0201/K(b) 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/K(b) or MAGE-3 expression in B16-HLA-MAGE-3. The ability of MAGE-3 antigen to be processed and presented in the B16-HLA-MAGE-3 cell line were observed by CTL activity detection and tumor challenge test.

RESULTS

Transcription and protein expression of HLA-A 0201/H-2k(b) and MAGE-3 were demonstrated in B16-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 processed 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.

摘要

目的

在HLA - A2.1转基因小鼠中建立肿瘤模型，以检测MAGE - 3疫苗的疗效，建立一种共表达HLA - A 0201/K(b)和MAGE - 3的细胞系。

方法

通过将HLA - A 0201/K(b)和MAGE - 3共转染至B16黑色素瘤细胞获得B16 - HLA - MAGE - 3黑色素瘤细胞。采用RT - PCR、流式细胞术（FCM）分析和蛋白质印迹法检测B16 - HLA - MAGE - 3中HLA - A 0201/K(b)或MAGE - 3表达的mRNA或蛋白质。通过细胞毒性T淋巴细胞（CTL）活性检测和肿瘤攻击试验观察MAGE - 3抗原在B16 - HLA - MAGE - 3细胞系中被加工和呈递的能力。