Wheat Flour Subjected to Microbial Transglutaminase Enzymatic Treatment in the Presence of Lysine Ethyl Ester for Alimentary Use in the Treatment of Celiac Disease.
试验速览
- 阶段
- 2 期
- 发起方
- 入组人数
- 20
- 试验地点
- 2
- 主要终点
- Pathologic variations in histologic analysis from baseline
研究概览
简要总结
Celiac disease is one of the most common forms of food intolerance (prevalence 1/200). The disease occurs in genetically predisposed individuals after ingestion of foods containing gluten. Celiac patients can suffer from severe malabsorption syndrome, mainly characterized by diarrhea and weight loss. The only therapeutic approach currently recognized is a life-long gluten-free diet.
Specific regions of gluten molecule become recognizable by lymphocytes and activate them, due to changes made by tissue transglutaminase. These changes consist in the conversion of specific residues of glutamine into glutamic acid. The consequence is an increased binding affinity between gluten and histocompatibility molecule (HLA-DQ2), localized on the surface of the "antigen presenting cells" (APC); the exposure of the fragments of modified gluten on the surface of APC is a phenomenon that eventually activates T lymphocytes.
Recent studies on modified gluten confirmed the hypothesis that it is possible to block the presentation of gluten to lymphocytes by means of lysine ethyl ester binding exclusively to those gluten regions responsible for lymphocyte activation.
The enzymatic treatment is performed directly on flour instead of extracted gluten, maintaining the same anti-inflammatory effectiveness.
The procedure uses a food-grade enzyme, the microbial transglutaminase (mTGasi) isolated from Streptoverticillium mobarensis, able to catalyze the formation of intermolecular "cross-links" that modify the functional properties of the products.
Objective of the study is to validate the ability of the enzyme treatment of wheat flour with mTGasi and lysine ethyl ester to block the toxic effect of gluten in celiac patients.
详细描述
Celiac disease is one of the most common forms of food intolerance (prevalence 1/200). The disease occurs in genetically predisposed individuals after ingestion of foods containing wheat gluten and similar proteins found in other common cereals such as barley and rye. Celiac patients can suffer from severe malabsorption syndrome, mainly characterized by diarrhea, weight loss and growth retardation. The only therapeutic approach currently recognized is a life-long gluten-free diet.
Specific regions of gluten molecule become recognizable by lymphocytes and activate them, due to changes made by tissue transglutaminase. These changes consist in the conversion of specific residues of glutamine (Q) into glutamic acid (E). The consequence is an increased binding affinity between gluten and histocompatibility molecule (HLA-DQ2), localized on the surface of the "antigen presenting cells" (APC); the exposure of the fragments of modified gluten on the surface of APC is a phenomenon that eventually activates T lymphocytes.
Recent studies on modified gluten confirmed the hypothesis that it is possible to block the presentation of gluten to lymphocytes by means of lysine ethyl ester binding exclusively to those gluten regions responsible for lymphocyte activation.
Specifically, it is possible to perform the enzymatic treatment directly on flour instead of extracted gluten, maintaining the same anti-inflammatory effectiveness. The final procedure consists in dissolving the flour in water in the presence of appropriate concentrations of enzyme and lysine ethyl ester, maintaining the suspension in constant motion for two hours at room temperature.
The procedure uses a food-grade enzyme, the microbial transglutaminase (mTGasi) isolated from Streptoverticillium mobarensis, already used for the preparation of food. The mTgasi is able to catalyze the formation of intermolecular "cross-links" modifying the functional properties of the products through the aggregation and polymerization of proteins. The peculiar method identified by our laboratory reduces the possibility of cross-links between proteins: consequently, minimal changes involve the gluten structure and, consequently, the visco-elastic properties of the dough.
研究设计
- 研究类型
- Interventional
- 分配方式
- Randomized
- 干预模型
- Parallel
- 主要目的
- Treatment
- 盲法
- Triple (Participant, Care Provider, Investigator)
入排标准
- 年龄范围
- 18 Years 至 65 Years（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •INCLUSION CRITERIA
- •diagnosis of celiac disease according to ESPGHAN criteria: class III according to Marsh-Oberhuber classification, clear response to gluten-free diet, serological antiendomysium and antitransglutaminase antibodies positive results before GFD;
- •HLA DQ2-DQ8 positive results;
- •gluten-free diet from at least one year;
- •negative serology from at least 1 year;
排除标准
- •inflammatory bowel diseases;
- •infectious liver disease;
- •renal impairment.
结局指标
主要结局
Pathologic variations in histologic analysis from baseline
时间窗: After 3 months, or in case of serum anti-tTG IgA/EMA IgA positive results
Proof of duodenal mucosa histological alterations induced by celiac disease, consisting in altered (\<3:1) villous height/crypt depth ratio and increased (\>25) intraepithelial lymphocytes per 100 intestinal epithelial cells, according to Marsh-Oberhuber classification.
Change from baseline in IgA anti-tissue transglutaminase (anti-tTG) antibodies at 30 days
时间窗: After 1 month
Proof of any positivization of specific serological antibodies for celiac disease. Results quantified by an ELISA reader at 450 nm (A450nm) is expressed in U/mL and the antibody level 10 U/mL was used as a cutoff value to identify anti-tTG positive results.
Change from baseline in IgA anti-endomysium antibodies (EMA) at 30 days
时间窗: After 1 month
Test used to confirm serological activation of celiac disease. IgA EMA were searched in sera diluted 1:5 by indirect immunofluorescence analysis on cryostat sections of monkey esophagus. The results are expressed as ''positive/negative''.
Change from baseline in IgA anti-endomysium antibodies (EMA) at 60 days
时间窗: After 2 months
Test used to confirm serological activation of celiac disease. IgA EMA were searched in sera diluted 1:5 by indirect immunofluorescence analysis on cryostat sections of monkey esophagus. The results are expressed as ''positive/negative''.
Change from baseline in IgA anti-tissue transglutaminase (anti-tTG) antibodies at 60 days
时间窗: After 2 months
Proof of any positivization of specific serological antibodies for celiac disease. Results quantified by an ELISA reader at 450 nm (A450nm) is expressed in U/mL and the antibody level 10 U/mL was used as a cutoff value to identify anti-tTG positive results.
Change from baseline in IgA anti-tissue transglutaminase (anti-tTG) antibodies at 90 days
时间窗: After 3 months
Proof of any positivization of specific serological antibodies for celiac disease. Results quantified by an ELISA reader at 450 nm (A450nm) is expressed in U/mL and the antibody level 10 U/mL was used as a cutoff value to identify anti-tTG positive results.
Change from baseline in IgA anti-endomysium antibodies (EMA) at 90 days
时间窗: After 3 months
Test used to confirm serological activation of celiac disease. IgA EMA were searched in sera diluted 1:5 by indirect immunofluorescence analysis on cryostat sections of monkey esophagus. The results are expressed as ''positive/negative''.
次要结局
- Variation of asthenia from baseline at 30 days(After 1 month)
- Presence of diarrhea at baseline(At baseline)
- Presence of asthenia at baseline(At baseline)
- Variation of diarrhea from baseline at 30 days(After 1 month)
- Variation of bloating from baseline at 60 days(After 2 months)
- Variation of abdominal pain from baseline at 90 days(After 3 months)
- Presence of abdominal pain at baseline(At baseline)
- Variation of asthenia from baseline at 90 days(After 3 months)
- Presence of bloating at baseline(At baseline)
- Variation of bloating from baseline at 30 days(After 1 month)
- Variation of diarrhea from baseline at 60 days(After 2 months)
- Variation of asthenia from baseline at 60 days(After 2 months)
- Variation of bloating from baseline at 90 days(After 3 months)
- Variation of abdominal pain from baseline at 30 days(After 1 month)
- Variation of abdominal pain from baseline at 60 days(After 2 months)
- Variation of diarrhea from baseline at 90 days(After 3 months)
研究者
Antonio Picarelli
Medical Doctor
University of Roma La Sapienza