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首页> 外文期刊>BMC Biotechnology >Heterologous expression of human costimulatory molecule B7-2 and construction of B7-2 immobilized polyhydroxyalkanoate nanoparticles for use as an immune activation agent
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Heterologous expression of human costimulatory molecule B7-2 and construction of B7-2 immobilized polyhydroxyalkanoate nanoparticles for use as an immune activation agent

机译：人类共刺激分子B7-2的异源表达和B7-2固定化聚羟基链烷酸酯纳米颗粒的构建，以用作免疫激活剂

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Background Costimulation of T cells via costimulatory molecules such as B7 is important for eliciting cell-mediated antitumor immunity. Presenting costimulation molecules by immobilizing recombinant B7 on the surface of nanovectors is a novel strategy for complementary therapy. Polyhydroxyalkanoates (PHAs) are a family of biodegradable, non-toxic, biocompatible polyesters, which can be used as a nonspecific immobilizing matrix for protein presentation. Recombinant protein fusion with PHA granule binding protein phasin (PhaP) can be easily immobilized on the surface of PHA nanoparticles through hydrophobic interactions between PhaP and PHA, and therefore provides a low-cost protein presenting strategy. Results In this study, the extracellular domain of the B7-2 molecule (also named as CD86) was fused with PhaP at its N-terminal and heterogeneously expressed in recombinant Escherichia coli strain BL21 (DE3). The purified B7-2-PhaP protein was immobilized on the surface of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx)-based nanoparticles. Loading of 240 μg (3.2 pMol) of B7-2-PhaP protein per mg nanoparticles was achieved. Immobilized B7-2-PhaP on PHBHHx nanoparticles induced T cell activation and proliferation in vitro . Conclusions A PHA nanoparticle-based B7-2 costimulation molecule-presenting system was constructed. The PHA-based B7 presenting nanosystem provided costimulation signals to induce T cell activation and expansion in vitro . The B7-2-PhaP immobilized PHA nanosystem is a novel strategy for costimulation molecule presentation and may be used for costimulatory molecule complementary therapy.
机译：背景技术通过共刺激分子（例如B7）对T细胞进行共刺激对于引发细胞介导的抗肿瘤免疫至关重要。通过将重组B7固定在纳米载体的表面上来提供共刺激分子是互补疗法的一种新策略。聚羟基链烷酸酯（PHA）是一类可生物降解的，无毒的，生物相容性的聚酯，可以用作蛋白质呈递的非特异性固定基质。通过PhaP和PHA之间的疏水相互作用，可以很容易地将重组蛋白与PHA颗粒结合蛋白phasin（PhaP）融合，从而将其固定在PHA纳米颗粒的表面上，从而提供了一种低成本的蛋白呈递策略。结果在这项研究中，B7-2分子的胞外域（也称为CD86）在其N端与PhaP融合，并在重组大肠杆菌BL21（DE3）菌株中异源表达。纯化的B7-2-PhaP蛋白固定在基于聚（3-羟基丁酸酯-co-3-羟基己酸酯）（PHBHHx）的纳米颗粒的表面上。每mg纳米颗粒负载240μg（3.2 pMol）的B7-2-PhaP蛋白。固定在PHBHHx纳米粒子上的B7-2-PhaP诱导体外T细胞活化和增殖。结论构建了基于PHA纳米粒子的B7-2共刺激分子呈递系统。基于PHA的B7呈递纳米系统提供了共刺激信号来诱导T细胞的体外活化和扩增。 B7-2-PhaP固定化的PHA纳米系统是一种共刺激分子呈递的新策略，可用于共刺激分子互补疗法。

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