Waddell T E, Lingwood C A, Gyles C L
Department of Veterinary Microbiology and Immunology, Ontario Veterinary College, University of Guelph, Canada.
Infect Immun. 1996 May;64(5):1714-9. doi: 10.1128/iai.64.5.1714-1719.1996.
In pigs with edema disease, verotoxin 2e (VT2e) is produced in the intestine and transported to tissues, but neither the mechanism by which toxin passes through the intestine nor its failure to induce an enterotoxic reaction is understood. Binding of VT2e to pig intestine was examined by enzyme-linked immunosorbent assay involving microvillus membranes (MVM) and crude mucus; thin-layer chromatographic overlay immunoassay with total lipids extracted from MVM; and indirect immunofluorescence of toxin bound to thin sections of jejunum, ileum, and colon. VT2e bound significantly to MVM from pig jejunum and ileum but not to crude mucus. Verotoxin 2e-binding glycolipids, globotetraosylceramide and globotriaosylceramide, were detected by thin-layer chromatographic overlay immunoassay in extracts of MVM from jejunum and ileum. Indirect immunofluorescence showed that VT2e bound to vessels within the submucosa and muscularis mucosa of the jejunum, ileum, and colon and to enterocytes at the lower portion but not at the tips of villi in the jejunum and ileum. Receptors for VT2e are therefore present in the intestine of the pig, but their role in absorption of VT2e is unclear since intraintestinal inoculation of pigs with large quantities of VT2e does not result in edema disease. Previously reported lack of enterotoxicity of verotoxins in pig intestine may be explained by the absence of toxin receptors in the villus absorptive enterocytes.
在患有水肿病的猪中,志贺毒素2e(VT2e)在肠道中产生并运输到组织,但毒素穿过肠道的机制及其未能引发肠毒性反应的原因尚不清楚。通过涉及微绒毛膜(MVM)和粗黏液的酶联免疫吸附测定、用从MVM中提取的总脂质进行的薄层色谱覆盖免疫测定以及对与空肠、回肠和结肠薄片结合的毒素进行间接免疫荧光,研究了VT2e与猪肠道的结合情况。VT2e与猪空肠和回肠的MVM有显著结合,但与粗黏液无结合。通过薄层色谱覆盖免疫测定在空肠和回肠MVM提取物中检测到了与VT2e结合的糖脂、球四糖神经酰胺和球三糖神经酰胺。间接免疫荧光显示,VT2e与空肠、回肠和结肠黏膜下层和黏膜肌层内的血管以及空肠和回肠绒毛下部而非顶端的肠上皮细胞结合。因此,猪肠道中存在VT2e的受体,但它们在VT2e吸收中的作用尚不清楚,因为给猪肠道内接种大量VT2e不会导致水肿病。先前报道的志贺毒素在猪肠道中缺乏肠毒性可能是由于绒毛吸收性肠上皮细胞中缺乏毒素受体。