Gavin M A, Dere B, Grandea A G, Hogquist K A, Bevan M J
Department of Immunology, University of Washington, Seattle 98195.
Eur J Immunol. 1994 Sep;24(9):2124-33. doi: 10.1002/eji.1830240929.
We describe a novel method for screening large libraries of random peptides for T cell antigens. Two libraries were constructed, containing fixed amino acids representing the major histocompatibility complex (MHC) class I anchor residues for H-2Kb-restricted octamers and H-2Db-restricted nonamers. Peptides from the Kb-restricted library (KbL: SXIXFXXL) and the Db-restricted library (DbL: XXXXNXXXIM) specifically stabilize empty Kb and Db molecules, respectively. The libraries contain peptides that mimic several H-2b-restricted cytotoxic T lymphocyte epitopes, and 21 mimotopes for a Db-restricted H-Y epitope were isolated. A degenerate synthetic peptide of limited complexity containing the identified H-Y sequence motif was found to be similar to the natural H-Y epitope by reverse-phase high performance liquid chromatography analysis. This peptide is also capable of immunizing female mice against male splenocytes. Several applications for MHC-restricted peptide libraries are discussed.
我们描述了一种筛选用于T细胞抗原的大型随机肽文库的新方法。构建了两个文库，其中包含代表H-2Kb限制的八聚体和H-2Db限制的九聚体的主要组织相容性复合体（MHC）I类锚定残基的固定氨基酸。来自Kb限制文库（KbL：SXIXFXXL）和Db限制文库（DbL：XXXXNXXXIM）的肽分别特异性稳定空的Kb和Db分子。这些文库包含模拟几种H-2b限制的细胞毒性T淋巴细胞表位的肽，并且分离出了21个针对Db限制的H-Y表位的模拟表位。通过反相高效液相色谱分析发现，含有已鉴定的H-Y序列基序的有限复杂性的简并合成肽与天然H-Y表位相似。该肽也能够免疫雌性小鼠抵抗雄性脾细胞。讨论了MHC限制肽文库的几种应用。
