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首页> 外文期刊>Infection and immunity >Cell-invasive activity of epitope-tagged adenylate cyclase of Bordetella pertussis allows in vitro presentation of a foreign epitope to CD8+ cytotoxic T cells.
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Cell-invasive activity of epitope-tagged adenylate cyclase of Bordetella pertussis allows in vitro presentation of a foreign epitope to CD8+ cytotoxic T cells.

机译：百日咳博德特氏菌的表位标记的腺苷酸环化酶的细胞侵袭活性允许将外源表位体外呈递给CD8 +细胞毒性T细胞。

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摘要

The adenylate cyclase (AC) toxin (CyaA) of Bordetella pertussis has an invasive catalytic domain (AC domain) which penetrates the cytoplasmic membrane of a variety of eukaryotic cells and intoxicates them by unregulated synthesis of cyclic AMP. Previous work led to identification of five permissive sites in the AC domain at which heterologous peptides are accommodated without affecting its enzymatic properties. We have constructed a set of CyaA toxins tagged at these permissive sites by insertion of a CD8+ T-cell epitope, RPQASGVYMGNLTAQ, from the nucleoprotein of lymphocytic choriomeningitis virus. Introduction of the epitope at any of the five sites did not affect the capacity of the toxin to deliver its AC domain into target cells. Moreover, the toxin with the inserted epitope was shown to sensitize target cells for lysis by epitope-specific CD8+ cytotoxic T lymphocytes in vitro, showing that the tagged AC was processed for presentation of the lymphocytic choriomeningitis virus epitope in association with the major histocompatibility complex class I molecules. This finding indicates that by virtue of delivery of foreign epitopes into the antigen-presenting cells, purpose-designed recombinant CyaAs may be useful for induction of specific major histocompatibility complex class I-restricted cell-mediated immunity also in vivo.
机译：百日咳博德特氏菌的腺苷酸环化酶（AC）毒素（CyaA）具有侵入性催化结构域（AC结构域），该结构域穿透各种真核细胞的细胞质膜并通过环状AMP的无节制合成而使其中毒。先前的工作导致在AC域中五个容性位点的鉴定，其中容纳了异源肽而不影响其酶学性质。我们通过从淋巴细胞性脉络膜脑膜炎病毒的核蛋白中插入CD8 + T细胞表位RPQASGVYMGNLTAQ，构建了在这些允许位点标记的一组CyaA毒素。在五个位点中的任何一个位点引入表位都不会影响毒素将其AC结构域递送至靶细胞的能力。此外，已证明具有插入的表位的毒素在体外可通过表位特异性CD8 +细胞毒性T淋巴细胞敏化靶细胞，表明标记的AC已被加工用于呈递淋巴细胞性脉络膜脑膜炎病毒表位以及主要的组织相容性复合体。我分子。该发现表明，由于将外来抗原决定簇递送至抗原呈递细胞中，目的设计的重组CyaAs也可用于在体内也诱导特定的主要组织相容性复合物I类限制的细胞介导的免疫。

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