Dewar David H, Amato Massimo, Ellis H Julia, Pollock Emma L, Gonzalez-Cinca Nuria, Wieser Herbert, Ciclitira Paul J
Rayne Institute (KCL Division of Nutrition), St Thomas' Hospital, Lambeth Palace Road, London, UK.
Eur J Gastroenterol Hepatol. 2006 May;18(5):483-91. doi: 10.1097/00042737-200605000-00005.
The ability of the gliadin fraction of wheat gluten to exacerbate coeliac disease is well documented. We investigated the possible toxicity of high molecular weight glutenin subunits (HMW-GS) in coeliac disease in vitro using gluten-sensitive T cells, and in vivo with challenge studies in patient volunteers.
A mixture of four HMW-GS was chemically separated from wheat flour and checked for purity by HPLC, SDS-PAGE and ELISA. T-cell lines, grown up from small intestinal biopsies from coeliac patients (n=17), were tested for their reactivity to HMW-GS. Adults with coeliac disease and who were on a gluten-free diet (n=3) underwent in-vivo challenges with HMW-GS. Duodenal biopsies, taken prior to the challenge and at intervals up to 6 h afterwards, were assessed for morphology, intra-epithelial lymphocyte count, and interleukin 15 (IL-15) expression, by immunohistochemistry.
T-cell lines from 11 of 17 patients were stimulated by HMW-GS. There was a significant change in small intestinal morphology 4 h after commencing infusions with HMW-GS in all three subjects. For example villus height to crypt depth ratios were reduced in the three patients from 3.0+/-0.5 to 1.29+/-0.2, 2.53+/-0.7 to 0.81+/-0.6 and 3.0+/-0.7 to 1.85+/-0.3, P<0.0001 in all cases. There was increased expression of IL-15 in the small intestine from 2 h after the HMW-GS challenges.
Mixed HMW-GS stimulate T-cell lines from some coeliac patients and exacerbate coeliac disease in vivo, inducing expression, within 2 h, of IL-15. This suggests an innate immune response to these proteins.
小麦麸质中的麦醇溶蛋白成分会加剧乳糜泻,这一点已有充分记录。我们使用对麸质敏感的T细胞,在体外研究了高分子量麦谷蛋白亚基(HMW - GS)在乳糜泻中的潜在毒性,并在体内对患者志愿者进行了激发试验研究。
从小麦粉中化学分离出四种HMW - GS的混合物,并通过高效液相色谱法(HPLC)、十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)和酶联免疫吸附测定(ELISA)检测其纯度。对从乳糜泻患者(n = 17)的小肠活检样本中培养出的T细胞系,检测其对HMW - GS的反应性。患有乳糜泻且正在接受无麸质饮食的成年人(n = 3)接受了HMW - GS的体内激发试验。在激发试验前以及之后每隔一段时间直至6小时采集十二指肠活检样本,通过免疫组织化学评估其形态、上皮内淋巴细胞计数和白细胞介素15(IL - 15)表达。
17例患者中有11例的T细胞系受到HMW - GS刺激。在所有三名受试者中,开始输注HMW - GS 4小时后,小肠形态有显著变化。例如,三名患者的绒毛高度与隐窝深度之比从3.0±0.5降至1.29±0.2、从2.53±0.7降至0.81±0.6以及从3.0±0.7降至1.85±0.3,所有情况下P<0.0001。HMW - GS激发试验后2小时起,小肠中IL - 15的表达增加。
混合的HMW - GS刺激了一些乳糜泻患者的T细胞系,并在体内加剧了乳糜泻,在2小时内诱导IL - 15表达。这表明对这些蛋白质存在先天性免疫反应。