PEPTIDE BINDING TO THE MOST FREQUENT HLA-A CLASS-I ALLELES MEASURED BY QUANTITATIVE MOLECULAR-BINDING ASSAYS
PEPTIDE BINDING TO THE MOST FREQUENT HLA-A CLASS-I ALLELES MEASURED BY QUANTITATIVE MOLECULAR-BINDING ASSAYS
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DOI：
10.1016/0161-5890(94)90019-1
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发表时间：
1994-08-01
影响因子：
3.6
通讯作者：
KUBO, RT
中科院分区：
文献类型：
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作者：
SETTE, A;SIDNEY, J;KUBO, RT
描述了针对几种常见的人类 HLA-A 等位基因（A1、A2.1、A3、A11 和 A24）的定量测定，以测量确定的合成抗原肽和纯化的 MHC I 类分子的结合。在适当的条件下，放射性标记的肽与纯化的 MHC I 类分子的结合非常有效、高度特异性，并且似乎依赖于由关键锚定残基位置定义的肽的特定序列基序。测定的建立和优化表明，从 EBV 转化的 B 细胞系来源中分离出的 MHC I 类分子的相对较高比例能够结合外源添加的肽。对所有等位基因的斯卡查德分析产生 5-10% 的占用值。肽长度对结合亲和力的直接影响反映了严格的肽大小要求。肽-MHC I 类相互作用在整体亲和力和动力学行为方面与肽-MHC II 类相互作用表现出显着的相似性。肽-MHC I 类结合测定的免疫学相关性还可以通过测量一组先前描述的 HLA 限制性肽与其 HLA 限制性元件的亲和力来证明。在 91% (10/11) 的情况下，肽的结合亲和力为 50 nM 或更低，而在其余 9% (1/11) 的情况下，肽的结合亲和力在 50 至 500 nM 范围内。因此，这些数据首次定量估计了 HLA-A 结合亲和力的水平与人类多种免疫显性 CTL 表位相关。
Quantitative assays to measure the binding of defined synthetic antigenic peptides and purified MHC class I molecules are described for several common human HLA-A alleles (A1, A2.1, A3, A11 and A24). Under appropriate conditions, the binding of radiolabeled peptides to purified MHC class I molecules is very effective, highly specific, and appears to be dependent on the specific sequence motif of the peptide as defined by critical anchor residue positions. Establishment and optimization of the assay reveals that a relatively high fraction of the MHC class I molecules isolated from EBV transformed B cell line sources is capable of binding exogenously added peptide. Scatchard analysis for all alleles yields 5-10% occupancy values. There is a stringent peptide size requirement that is reflected by the direct influence of peptide length on the binding affinity. The peptide-MHC class I interactions demonstrate remarkable similarity to peptide-MHC class II interactions, both in overall affinity and kinetic behavior. The immunological relevance of the peptide-MHC class I binding assay is also demonstrated by measuring the affinity of a panel of previously described HLA restricted peptides for their HLA restriction element. In 91% (10/11) of the cases, the peptides bound with affinities of 50 nM or less, and in the remaining 9% (1/11) of the cases, in the 50 to 500 nM range. Thus, these data provide the first quantitative estimate of what level of HLA-A binding affinity is associated with a diverse panel of immunodominant CTL epitopes in man.