A constructed HLA-A2-restricted pMAGE-AI(278-286) tetramer detects specific cytotoxic T lymphocytes in tumour tissues in situ

作者：Mongkhoune, Sodaly; Xie, Yu-An; Wang, Yu-Qing; Chen, Yue; Zhou, Nuo; Peng, Yi; He, Jian; Yu, Xia; Zhou, Su-Fang; Luo, Guo-Rong; Zhao, Yong-Xiang; Lu, Xiao-Ling^*

来源：Journal of International Medical Research, 2013, 41(6): 1811-1824.

DOI：10.1177/0300060513496187

• Tumour-specific cytotoxic T lymphocytes
• HLA-A2-pMAGE-A1(278-286) tetramer
• in situ tetramer staining

Objective To construct a human leucocyte antigen (HLA)-A2-restricted peptide 278-286 of melanoma-associated antigen family A, 1 (pMAGE-A1(278-286)) tetramer to analyse the distribution of cytotoxic T lymphocytes (CTLs) in tumour tissue and tumour-adjacent normal tissue.
Methods A HLA-A2-pMAGE-A1(278-286) tetramer was constructed. The distribution of pMAGE-A1(278-286)-specific CTLs was investigated in tumour tissues and tumour-adjacent normal tissues from patients with hepatocellular carcinoma using in situ HLA-A2-pMAGE-A1(278-286) tetramer staining.
Results Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis indicated that HLA-A2 heavy and light chain proteins were successfully obtained. The successful construction of the HLA-A2-pMAGE-A1(278-286) monomer was confirmed with Western blot analysis using W6/32 antibody. Flow cytometry confirmed the specific binding of HLA-A2-pMAGE-A1(278-286) tetramer to pMAGE-A1(278-286)-specific CTLs. In situ HLA-A2-pMAGE-A1(278-286) tetramer staining demonstrated that the number of pMAGE-A1(278-286)-specific CTLs in tumour tissues was significantly higher than in tumour-adjacent normal tissues.
Conclusions The HLA-A2-pMAGE-A1(278-286) tetramer was useful for the detection of pMAGE-A1(278-286)-specific CTLs in both tumour tissues and tumour-adjacent normal tissues. In situ tetramer staining is a powerful tool for investigating the distribution of pMAGE-A1(278-286)-specific CTLs in the tumour microenvironment.

• 出版日期2013-12
• 单位广西医科大学