de Bueger M, Verreck F, Blokland E, Drijfhout J W, Amons R, Koning F, Goulmy E
Department of Immunohaematology, University Hospital Leiden, The Netherlands.
Eur J Immunol. 1993 Mar;23(3):614-8. doi: 10.1002/eji.1830230305.
Purified HLA-A2.1 molecules obtained by affinity chromatography of 6 x 10(10) Epstein Barr virus (EBV)-transformed B lymphocytes were used in an attempt to isolate the human HLA-A2.1-restricted minor histocompatibility (H) peptides H-Y and HA-2. Fraction 18 of the high-performance liquid chromatography (HPLC)-separated HLA-A2.1 peptide pool was found to contain the natural HA-2 peptide. An HA-2-specific, HLA-A2.1-restricted cytotoxic T lymphocyte clone lysed HLA-A2.1+ HA-2- EBV-transformed B lymphocyte cell lines reproducibly and in a concentration-dependent fashion in the presence of fraction 18, but not in the presence of other HPLC fractions. By contrast, H-Y sensitizing activity was not found in any fraction. Amino acid sequencing of peptide fraction 18 revealed a mixture of peptides with maximal length of nine amino acids, in which the presence of Leu at positions 2 and 9 was dominant. Surprisingly, the HA-2 peptide could not be mimicked by any of the peptide mixtures synthesized according to the amino acid sequences found in fraction 18. Our failure to obtain the actual amino acid sequence of the human minor H peptide HA-2 from a peptide pool with the established pattern for binding to HLA-A2.1 may indicate that this CTL defined minor H peptide does not represent an abundant HLA-A2.1 binding peptide.
通过对6×10¹⁰个爱泼斯坦-巴尔病毒（EBV）转化的B淋巴细胞进行亲和层析获得的纯化HLA - A2.1分子，被用于尝试分离人类HLA - A2.1限制性次要组织相容性（H）肽H - Y和HA - 2。发现高效液相色谱（HPLC）分离的HLA - A2.1肽库的第18部分含有天然HA - 2肽。一个HA - 2特异性、HLA - A2.1限制性细胞毒性T淋巴细胞克隆，在第18部分存在的情况下，可重复性地以浓度依赖方式裂解HLA - A2.1⁺HA - 2⁻EBV转化的B淋巴细胞系，但在其他HPLC部分存在时则不能。相比之下，在任何部分都未发现H - Y致敏活性。肽第18部分的氨基酸测序揭示了最大长度为九个氨基酸的肽混合物，其中第2位和第9位存在亮氨酸占主导。令人惊讶的是，根据第18部分中发现的氨基酸序列合成的任何肽混合物都无法模拟HA - 2肽。我们未能从具有与HLA - A2.1结合既定模式的肽库中获得人类次要H肽HA - 2的实际氨基酸序列，这可能表明这种由细胞毒性T淋巴细胞定义的次要H肽并不代表一种丰富的HLA - A2.1结合肽。
