Anjuère F, Horvath C, Cerottini J C, Luescher I F
Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne, Epalinges, Switzerland.
Eur J Immunol. 1995 Jun;25(6):1535-40. doi: 10.1002/eji.1830250610.
The identification of endogenously produced antigenic peptides presented by MHC class I molecules has opened the way to peptide-based strategies for CTL induction in vivo. Here we demonstrate that the induction in vivo of CTL directed against naturally processed antigens can be triggered by injection of syngeneic cells expressing covalent major histocompatibility complex class I-peptide complexes. In the model system used, the induction of HLA-Cw3 specific cytotoxic T lymphocytes (CTL) in mice by cell surface-associated, covalent H-2Kd (Kd)-Cw3 peptide complexes was investigated. The Kd-restricted Cw3 peptide 170-179 (RYLKNGKETL), which mimics the major natural epitope recognized by Cw3-specific CTL in H-2d mice, was converted to a photoreactive derivative by replacing Arg-170 with N-beta-(4-azidosalicyloyl)-L-2,3-diaminopropionic acid. This peptide derivative was equivalent to the parental Cw3 peptide in terms of binding to Kd molecules and recognition by Cw3-specific CTL clones and could be cross-linked efficiently and selectively to Kd molecules on the surface of Con A-stimulated spleen cells from H-2d mice. Photocross-linking prevented the rapid dissociation of Kd-peptide derivative complexes that takes place under physiological conditions. Cultures of spleen cells or peritoneal exudate cells from mice inoculated i.p. with peptide-pulsed and photocross-linked cells developed a strong CTL response following antigenic stimulation in vitro. The cultured cells efficiently lysed not only target cells sensitized with the Cw3 170-179 peptide but also target cells transfected with the Cw3 gene. Moreover, their TCR preferentially expressed V beta 10 and J alpha pHDS58 segments as well as conserved junctional sequences, as has been observed previously in Cw3-specific CTL responses. In contrast, no Cw3-specific CTL response could be obtained in cultures derived from mice injected with Con A-stimulated spleen cells pulsed with the peptide derivative without photocross-linking.
MHC I类分子呈递的内源性产生的抗原肽的鉴定为体内诱导CTL的基于肽的策略开辟了道路。在此，我们证明通过注射表达共价主要组织相容性复合体I类 - 肽复合物的同基因细胞，可以在体内触发针对天然加工抗原的CTL诱导。在所使用的模型系统中，研究了细胞表面相关的共价H - 2Kd（Kd） - Cw3肽复合物在小鼠中诱导HLA - Cw3特异性细胞毒性T淋巴细胞（CTL）的情况。Kd限制性Cw3肽170 - 179（RYLKNGKETL）模拟了H - 2d小鼠中Cw3特异性CTL识别的主要天然表位，通过用N - β - （4 - 叠氮水杨酰基） - L - 2,3 - 二氨基丙酸取代Arg - 170将其转化为光反应性衍生物。就与Kd分子的结合以及被Cw3特异性CTL克隆识别而言，该肽衍生物与亲本Cw3肽相当，并且可以有效且选择性地与来自H - 2d小鼠的Con A刺激的脾细胞表面的Kd分子交联。光交联阻止了在生理条件下发生的Kd - 肽衍生物复合物的快速解离。经腹腔接种肽脉冲和光交联细胞的小鼠的脾细胞或腹腔渗出细胞培养物在体外抗原刺激后产生了强烈的CTL反应。培养的细胞不仅有效地裂解了用Cw3 170 - 179肽致敏的靶细胞，还裂解了用Cw3基因转染的靶细胞。此外，它们的TCR优先表达Vβ10和JαpHDS58片段以及保守的连接序列，如先前在Cw3特异性CTL反应中所观察到的。相比之下，在用未进行光交联的肽衍生物脉冲的Con A刺激的脾细胞注射的小鼠来源的培养物中，未获得Cw3特异性CTL反应。
