Laboratorio de Fisiopatogenia, Departamento de Fisiología, Instituto de Fisiología y Biofísica Bernardo Houssay (IFIBIO Houssay-CONICET), Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, 1121 Buenos Aires, Argentina.
Laboratorio de Patogénesis e Inmunología de Procesos Infecciosos, Instituto de Medicina Experimental (IMEX-CONICET), Academia Nacional de Medicina, Pacheco de Melo 3081, 1425 Buenos Aires, Argentina.
Vaccine. 2018 Mar 20;36(13):1728-1735. doi: 10.1016/j.vaccine.2018.02.060. Epub 2018 Feb 23.
E. coli O157:H7 is a foodborne pathogen responsible for bloody diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS). The objective of the present work was to evaluate the ability of colostral IgG obtained from Stx2-immunized cows to prevent against E. coli O157:H7 infection and Stx2 cytotoxicity. Hyperimmune colostrum (HC) was obtained from cows intramuscularly immunized with inactivated Stx2 or vehicle for controls. Colostral IgG was purified by affinity chromatography. Specific IgG antibodies against Stx2 and bovine lactoferrin (bLF) levels in HC and the corresponding IgG (HC-IgG/bLF) were determined by ELISA. The protective effects of HC-IgG/bLF against Stx2 cytotoxicity and adhesion of E. coli O157:H7 and its Stx2-negative mutant were analyzed in HCT-8 cells. HC-IgG/bLF prevention against E. coli O157:H7 was studied in human colon and rat colon loops. Protection against a lethal dose of E. coli O157:H7 was evaluated in a weaned mice model. HC-IgG/bLF showed high anti-Stx2 titers and high bLF levels that were able to neutralize the cytotoxic effects of Stx2 in vitro and in vivo. Furthermore, HC-IgG/bLF avoided the inhibition of water absorption induced by E. coli O157:H7 in human colon and also the pathogenicity of E. coli O157:H7 and E. coli O157:H7Δstx2 in rat colon loops. Finally, HC-IgG/bLF prevented in a 100% the lethality caused by E. coli O157:H7 in a weaned mice model. Our study suggests that HC-IgG/bLF have protective effects against E. coli O157:H7 infection. These beneficial effects may be due to specific anti-Stx2 neutralizing antibodies in combination with high bLF levels. These results allow us to consider HC-IgG/bLF as a nutraceutical tool which could be used in combination with balanced supportive diets to prevent HUS. However further studies are required before recommendations can be made for therapeutic and clinical applications.
产志贺毒素 2 型大肠杆菌(E. coli O157:H7)是一种食源性致病菌,可引起血性腹泻、出血性结肠炎和溶血尿毒综合征(HUS)。本研究旨在评估从 Stx2 免疫奶牛获得的初乳 IgG 预防 E. coli O157:H7 感染和 Stx2 细胞毒性的能力。通过肌肉内免疫灭活 Stx2 或对照载体获得高免初乳(HC)。通过亲和层析纯化初乳 IgG。通过 ELISA 测定 HC 中针对 Stx2 和牛乳铁蛋白(bLF)的特异性 IgG 抗体水平及其相应的 IgG(HC-IgG/bLF)。分析了 HC-IgG/bLF 对 HCT-8 细胞中 E. coli O157:H7 及其 Stx2 阴性突变体的粘附和 Stx2 细胞毒性的保护作用。在人结肠和大鼠结肠环中研究了 HC-IgG/bLF 对 E. coli O157:H7 的预防作用。在断奶小鼠模型中评估了 HC-IgG/bLF 对致死剂量 E. coli O157:H7 的保护作用。HC-IgG/bLF 显示出高抗 Stx2 效价和高 bLF 水平,能够中和 Stx2 的体外和体内细胞毒性作用。此外,HC-IgG/bLF 避免了 E. coli O157:H7 诱导的人结肠水分吸收抑制,也避免了 E. coli O157:H7 和 E. coli O157:H7Δstx2 在大鼠结肠环中的致病性。最后,HC-IgG/bLF 在断奶小鼠模型中 100%预防了 E. coli O157:H7 引起的致死性。本研究表明,HC-IgG/bLF 对 E. coli O157:H7 感染具有保护作用。这些有益作用可能是由于特异性抗 Stx2 中和抗体与高 bLF 水平的结合。这些结果使我们认为 HC-IgG/bLF 是一种营养保健品,可与均衡的支持性饮食联合使用,以预防 HUS。然而,在提出治疗和临床应用建议之前,还需要进一步的研究。