Münz C, Obst R, Osen W, Stevanović S, Rammensee H G
Department of Immunology, Institute for Cell Biology, University of Tübingen, Germany.
J Immunol. 1999 Jan 1;162(1):25-34.
PBL from HLA-A2- or HLA-A3- donors were stimulated with synthetic peptide libraries fitting HLA-A2 or HLA-A3 motifs and presented on HLA-A2- or HLA-A3-expressing TAP- cells. Peptide library-specific allorestricted CTL were found to constitute up to half the alloreactive CTL response and occurred at twofold lower frequency than autologous peptide library-specific CTL. This indicates that positive selection by one particular MHC class I molecule is not absolutely essential for the generation of CTL restricted to the same molecule. However, positive selection increases their frequency. The CTL obtained differed greatly both with respect to peptide dependency and peptide specificity. Determination of the peptide avidity for one representative CTL clone, 10F4, proved that the method described here allows the stimulation of high avidity cytotoxic T cells. This approach involving in vitro stimulation of T cells restricted toward a MHC molecule that was not present during their negative selection might therefore offer the possibility of isolating CTL against self and foreign peptides with varying avidities. Such T cells might indeed be useful for tumor immunotherapy.
来自HLA - A2或HLA - A3供体的外周血淋巴细胞（PBL）用符合HLA - A2或HLA - A3基序的合成肽库进行刺激，并呈递给表达HLA - A2或HLA - A3的TAP缺陷细胞。发现肽库特异性同种异体限制性细胞毒性T淋巴细胞（CTL）构成了高达一半的同种异体反应性CTL应答，且其出现频率比自体肽库特异性CTL低两倍。这表明由一种特定的I类主要组织相容性复合体（MHC）分子进行的阳性选择对于产生受同一分子限制的CTL并非绝对必要。然而，阳性选择会增加它们的频率。所获得的CTL在肽依赖性和肽特异性方面差异很大。对一个代表性的CTL克隆10F4的肽亲和力测定证明，此处描述的方法能够刺激高亲和力的细胞毒性T细胞。因此，这种涉及体外刺激在阴性选择期间不存在的针对MHC分子的T细胞的方法，可能提供分离针对具有不同亲和力的自身和外源肽的CTL的可能性。这样的T细胞可能确实对肿瘤免疫治疗有用。
