Schrama D, Andersen M H, Terheyden P, Schrøder L, Pedersen L O, thor Straten P, Becker J C
Department of Dermatology, Julius Maximilians-University, Würzburg, Germany.
Cancer Res. 2001 Jan 15;61(2):493-6.
Ex vivo ELISPOT analysis of peripheral blood lymphocytes obtained from stage IV melanoma patients demonstrated reactivity against peptides derived from MART-1 and gp100. However, the number of reactive T cells was < 1% that of total lymphocytes as detected by flow cytometry using tetrameric MHC/peptide complexes. Despite this low frequency, we were able to directly isolate these populations ex vivo by means of magnetic beads coated with MHC/peptide complexes and to subject these cells to T-cell receptor clonotype mapping. This analysis revealed that the MART-1/A0201- and gp100/A0201-reactive T-cell populations are composed of oligoclonal T cells that engage several T-cell receptor beta chain families. Longitudinal studies using this approach may result in a better correlation between T-cell reactivity and the course of neoplastic disease.
对IV期黑色素瘤患者外周血淋巴细胞进行的体外ELISPOT分析表明，其对源自MART-1和gp100的肽具有反应性。然而，通过使用四聚体MHC/肽复合物的流式细胞术检测，反应性T细胞的数量不到总淋巴细胞数量的1%。尽管频率较低，但我们能够通过包被有MHC/肽复合物的磁珠在体外直接分离这些细胞群，并对这些细胞进行T细胞受体克隆型图谱分析。该分析表明，MART-1/A0201反应性和gp100/A0201反应性T细胞群由参与多个T细胞受体β链家族的寡克隆T细胞组成。使用这种方法进行的纵向研究可能会使T细胞反应性与肿瘤疾病进程之间具有更好的相关性。
