Laboratory of Glycobiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla St. 12, 53-114, Wroclaw, Poland.
Biochem Biophys Res Commun. 2024 Dec 20;739:150975. doi: 10.1016/j.bbrc.2024.150975. Epub 2024 Nov 12.
Shiga toxin types 1 (Stx1) and 2 (Stx2), produced by Shiga toxin-producing Escherichia coli (STEC) and Shigella dysenteriae, are key virulence factors responsible for severe foodborne diseases, such as hemorrhagic colitis and hemolytic uremic syndrome (HUS). The receptors for Stxs are Gb3 and P1 glycotope, which contain the Galα1→4Gal epitope and are synthesized by human α1,4-galactosyltransferase (A4galt). Stx-related infections pose a global public health challenge, owing to the limited therapeutic options due to the restricted use of antibiotics. Therefore, there is an urgent need to develop novel therapeutic strategies. This study proposes an innovative strategy utilizing exosomes derived from CHO-Lec2 cells, which were modified with Functional-Spacer-Lipid (FSL) conjugates bearing the Gb3 carbohydrate epitope (exo-Gb3-FSL). Flow cytometry analysis confirmed the presence of Galα1→4Gal disaccharides on exo-Gb3-FSL constructs, enabling them to bind Stx1. Moreover, using CHO-Lec2 cells evaluated the ability of exo-Gb3-FSL agents to bind Stx1 and protect these cells from Stx1-mediated cytotoxicity. For Stx1-treated CHO-Lec2 cells, increased cell survival was observed when using 25 μM exo-Gb3-FSL constructs, compared to control cells. These findings highlight the potential of exosome-based anti-Stx1 agents as promising alternatives to conventional therapies. This innovative strategy may provide novel directions for studies on Stx1 neutralization, offering a valuable strategy for the treatment of Stx-related diseases.
志贺毒素 1 型（Stx1）和 2 型（Stx2）由产志贺毒素大肠杆菌（STEC）和痢疾志贺菌产生，是导致严重食源性疾病的关键毒力因子，如出血性结肠炎和溶血尿毒综合征（HUS）。Stx 的受体是 Gb3 和 P1 糖基表位，它们含有 Galα1→4Gal 表位，由人类 α1,4-半乳糖基转移酶（A4galt）合成。由于抗生素的使用受限，导致治疗选择有限，Stx 相关感染对全球公共卫生构成挑战。因此，迫切需要开发新的治疗策略。本研究提出了一种利用 CHO-Lec2 细胞衍生的外泌体的创新策略，该外泌体经过修饰，带有携带 Gb3 碳水化合物表位的功能性间隔脂质（FSL）缀合物（exo-Gb3-FSL）。流式细胞术分析证实了 exo-Gb3-FSL 构建体上存在 Galα1→4Gal 二糖，使其能够与 Stx1 结合。此外，使用 CHO-Lec2 细胞评估了 exo-Gb3-FSL 试剂结合 Stx1 的能力，并保护这些细胞免受 Stx1 介导的细胞毒性。对于用 Stx1 处理的 CHO-Lec2 细胞，与对照细胞相比，当使用 25 μM exo-Gb3-FSL 构建体时，观察到细胞存活率增加。这些发现突出了基于外泌体的抗 Stx1 试剂作为传统疗法的有前途替代品的潜力。这种创新策略可能为 Stx1 中和研究提供新的方向，为 Stx 相关疾病的治疗提供有价值的策略。
