ABSTRACT
Wheat-dependent exercise-induced anaphylaxis (WDEIA) is one of the most severe forms of wheat allergy. It occurs in patients when they exercise after ingesting wheat-containing foods. Nowadays, the only possible alternative for WDEIA patients is to avoid such foods. This study investigated the potential of six RNA of interference (RNAi) wheat lines with low-prolamin content as alternatives for WDEIA patients. For that purpose, a high performance-liquid chromatography (HPLC) analysis was performed to evaluate differences in gluten protein fractions among these lines. Next, western blots were conducted to measure the immunoglobulin E (IgE) reactivity to wheat proteins in sera from five WDEIA patients. Additionally, monoclonal antibodies (moAb) recognition sites and the IgE binding sites were searched in all peptides identified by LC-MS/MS after protein digestion. The results showed a 61.4%-81.2% reduction in the gliadin content of the RNAi lines, accompanied by an increase in their high-molecular weight (HMW) glutenin content compared to the wild type bread wheat line (WT). In all cases, the reduction in gliadin content correlated with a decrease in IgE reactivity observed in the sera of WDEIA patients, highlighting the E82 and H320 lines. These two RNAi lines exhibited a ≤90% reduction in IgE reactivity. This reduction could be attributed to an absence of IgE binding sites associated with α- and ω5-gliadins, which were present in the WT. Overall, these lines offer a potential alternative for foodstuff for individuals with WDEIA.
ABSTRACT
INTRODUCCIÓN: la enfermedad celiaca (EC) presenta un determinado patrón de infiltrado linfocitario en la mucosa duodenal. Gracias a la citometría de flujo como herramienta complementaria al diagnóstico de la EC, se pueden cuantificar y caracterizar los linfocitos intraepiteliales (LIE) a través de lo que comúnmente denominamos linfograma. Con este estudio, pretendemos describir nuestra experiencia con la técnica en el diagnóstico del paciente celiaco adulto. MÉTODOS: se han analizado retrospectivamente los linfogramas realizados en nuestro centro entre 2009 y 2017, que fueron en total 157. Catorce de ellos tenían un diagnóstico previo de EC y seguían una dieta sin gluten, 21 tuvieron un diagnóstico nuevo de EC y el resto fueron considerados no celiacos. Se ha estudiado la asociación de los valores del linfograma (LIE totales, linfocitos CD3- y linfocitos TcRγδ) con el diagnóstico de EC, el cumplimiento de la dieta sin gluten (DSG), el tiempo desde el diagnóstico y el título de inmunoglobulina A antitransglutaminasa tisular. RESULTADOS: el valor de área bajo la curva ROC de los linfocitos TcRγδ para el diagnóstico de EC varía entre 0,86 y 0,86. El porcentaje de linfocitos TcRγδ en pacientes celiacos en DSG es menor 12 (8,5) vs. 20,5 (8,7), p = 0,0153, aunque permanece elevado frente a los no celiacos 12 (8,5) vs. 6,7 (6), p = 0,135. El tiempo desde el diagnóstico y el título de IgA antitransglutaminasa tisular (anti-Tgt) se correlacionan con los valores del linfograma en el paciente celiaco. La infección por Helicobacter pylori y el tratamiento con antagonistas de receptores de angiotensina 2 (ARA2) se asocia a diferencias en el linfograma. CONCLUSIONES: el linfograma duodenal es una herramienta complementaria fiable en el diagnóstico del celiaco adulto. Sin embargo, el cumplimiento y la duración de la DSG, así como otros factores, pueden condicionar su capacidad diagnóstica
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Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Celiac Disease/diagnosis , Flow Cytometry , Lymphocytes/pathology , Helicobacter pylori , Helicobacter Infections , Retrospective Studies , ROC CurveABSTRACT
INTRODUCTION: celiac disease (CD) patients have a specific pattern of lymphocytic infiltrate in the duodenal mucosa. Flow cytometry is a complementary tool for the diagnosis of CD, which allows the quantification and characterization of intraepithelial lymphocytes (IELs) by what is commonly called a lymphogram. Here we describe our experience with this technique in the diagnosis of CD in adult patients. METHODS: lymphograms from 157 patients performed in our center between 2009 and 2017 were retrospectively analyzed. Fourteen patients had a previous diagnosis of CD and followed a gluten-free diet (GFD), 21 had a new diagnosis of CD and the remaining were considered as non-celiac. The association of the lymphogram results (total IELs, CD3- lymphocytes and TcRγδ lymphocytes) with the CD diagnosis, compliance with the GFD, time since diagnosis and IgA anti-TG2 titer were determined. RESULTS: the area under the ROC curve of TcRγδ lymphocytes for CD patients varied between 0.86 and 0.86. The percentage of TcRγδ lymphocytes in GFD-treated patients was lower; 12 (8.5) vs 20.5 (8.7), p = 0.0153. However, it remained high compared to non-CD; 12 (8.5) vs 6.7 (6), p = 0.135. The time since diagnosis and IgA anti-TG2 titer correlated with the lymphogram results. Helicobacter pylori infection and treatment with angiotensin receptor antagonist 2 (ARA2) were associated with differences in the lymphogram results in patients without CD. CONCLUSIONS: the duodenal lymphogram is a reliable complementary tool in adults for the diagnosis of CD. However, compliance and duration of the GFD and other factors may condition its diagnostic capacity.
