Celiac disease-associated Neisseria flavescens decreases mitochondrial respiration in CaCo-2 epithelial cells: Impact of Lactobacillus paracasei CBA L74 on bacterial-induced cellular imbalance.

We previously identified a Neisseria flavescens strain in the duodenum of celiac disease (CD) patients that induced immune inflammation in ex vivo duodenal mucosal explants and in CaCo-2 cells. We also found that vesicular trafficking was delayed after the CD-immunogenic P31-43 gliadin peptide-entered CaCo-2 cells and that Lactobacillus paracasei CBA L74 (L. paracasei-CBA) supernatant reduced peptide entry. In this study, we evaluated if metabolism and trafficking was altered in CD-N. flavescens-infected CaCo-2 cells and if any alteration could be mitigated by pretreating cells with L. paracasei-CBA supernatant, despite the presence of P31-43. We measured CaCo-2 bioenergetics by an extracellular flux analyser, N. flavescens and P31-43 intracellular trafficking by immunofluorescence, cellular stress by TBARS assay, and ATP by bioluminescence. We found that CD-N. flavescens colocalised more than control N. flavescens with early endocytic vesicles and more escaped autophagy thereby surviving longer in infected cells. P31-43 increased colocalisation of N. flavescens with early vesicles. Mitochondrial respiration was lower (P < .05) in CD-N. flavescens-infected cells versus not-treated CaCo-2 cells, whereas pretreatment with L. paracasei-CBA reduced CD-N. flavescens viability and improved cell bioenergetics and trafficking. In conclusion, CD-N. flavescens induces metabolic imbalance in CaCo-2 cells, and the L. paracasei-CBA probiotic could be used to correct CD-associated dysbiosis.

Effect of pH control during rice fermentation in preventing a gliadin P31-43 entrance in epithelial cells.

Coeliac disease is an increasingly recognised pathology, induced by the ingestion of gluten in genetically predisposed patients. Undigested gliadin peptide can induce adaptive and innate immune response that unleash the typical intestinal mucosal alterations. A growing attention is paid to alternative therapeutic approaches to the gluten-free diet: one of these approaches is the use of probiotics and/or postbiotics. We performed lactic fermentation of rice flour with and without pH control, using Lactobacillus paracasei CBA L74 as fermenting strain. We evaluated bacterial growth, lactic acid production during fermentation and gliadin peptide P31-43 entrance in CaCo-2 cells with and without pH control. When pH control was applied no differences were observed in terms of bacterial growth; on the contrary, lactic acid production was greater, as expected. Both samples could inhibit the P31-43 entrance in CaCo-2 cells but the effect was significantly greater for samples obtained when the pH control was applied.

Supplementation of Reduced Gluten Barley Diet with Oral Prolyl Endopeptidase Effectively Abrogates Enteropathy-Associated Changes in Gluten-Sensitive Macaques.

Celiac disease (CD) is an autoimmune disorder that affects approximately three million people in the United States. Furthermore, non-celiac gluten sensitivity (NCGS) affects an estimated additional 6% of the population, e.g., 20 million in the U.S. The only effective treatment of CD and NCGS requires complete removal of gluten sources from the diet. While required adherence to a gluten-free diet (GFD) is extremely difficult to accomplish, efforts to develop additional supportive treatments are needed. To facilitate these efforts, we developed a gluten-sensitive (GS) rhesus macaque model to study the effects of novel therapies. Recently reported results from phase one of this project suggest that partial improvement-but not remission-of gluten-induced disease can be accomplished by 100-fold reduction of dietary gluten, i.e., 200 ppm-by replacement of conventional dietary sources of gluten with a mutant, reduced gluten (RG) barley (lys3a)-derived source. The main focus of this (phase two) study was to determine if the inflammatory effects of the residual gluten in lys3a mutant barley grain could be further reduced by oral supplementation with a prolylendopeptidase (PE). Results reveal that PE supplementation of RG barley diet induces more complete immunological, histopathological and clinical remission than RG barley diet alone. The combined effects of RG barley diet and PE supplementation resulted in a further decrease of inflammatory mediators IFN-γ and TNF secretion by peripheral lymphocytes, as well as decreased plasma anti-gliadin and anti-intestinal tissue transglutaminase (TG2) antibodies, diminished active caspase production in small intestinal mucosa, and eliminated clinical diarrhea-all comparable with a gluten-free diet induced remission. In summary, the beneficial results of a combined RG barley and PE administration in GS macaques may warrant the investigation of similar synergistic approaches.

Oat raw materials and bakery products - amino acid composition and celiac immunoreactivity.

BACKGROUND: The aim of this study was to compare the biochemical and immunochemical properties of avenins in some special oat raw materials and additionally the possibility of using them as a raw material for the gluten-free bakery products. METHODS: The compared oat raw materials were - oat flakes, commercial oat flours (including gluten-free oat flour) and residual oat flour, which is by-product of ß-glucan preparation. Biochemical characteristic included amino acid compositions and SDS-PAGE profiles of extracted avenins. The immunochemical reactivity with polyclonal anti-gluten and monoclonal anti-gliadin antibodies was evaluated qualitatively and quantitatively by immunoblotting and ELISA methods. Additionally, experimental bakery products made of examined raw materials were assessed according to their suitability for the celiac patients' diet. RESULTS: The highest protein content was measured in the ß-glucan preparation "Betaven" and gluten-free oat flour. Proteins of all materials are rich in glutamic and aspartic acid, leucine and arginine. Proportions of amino acids in avenins extracted from most of oat raw materials are similar, excluding gluten-free oat flour, which has a very low avenin content and proportions of individual amino acids are different. The SDS-PAGE protein pattern consisted of proteins with molecular weight of about 25-35 kDa. Polyclonal anti-gluten anti-body recognized all protein fractions of molecular weight higher than 20 kDa. Quantitative ELISA analysis shows that the majority of samples has a gliadin-like protein content within the range of 80-260 mg/kg, excluding gluten-free flours and corresponding bakery products. Altogether, ß-glucan preparation has extremely high level of gliadin-like proteins. CONCLUSIONS: In the examined oat raw materials and foods the contents of immunoreactive amino acid sequences exceeded the limit of 20 mg/kg (considered as gluten-free) except for gluten-free flours (oat and  the prepared mixture) and the bakery products based on gluten-free flours. Unfortunately, the rest of oat raw materials and products cannot be considered gluten-free.

Is there any relationship between unrecognized Celiac disease and unexplained infertile couples?

BACKGROUND/AIMS: Celiac Disease (CD) is a chronic autoimmune disease characterized by small intestinal malabsorbtion and diarrhea, triggered by the ingestion of food products containing gluten. There are studies reporting that some nutritional deficiencies and some factors related to immunity may cause a decrease in fertility as well as some problems in sperm parameters. The prevalence of CD in unexplained infertility (UEI) couples is not as high as that mentioned in some reports. There is no accurate knowledge about the prevalence of CD in a UEI couple. MATERIALS AND METHODS: A total of 68 couples with UEI who were admitted at Türk Diyanet Vakfi 29 Mayis Hospital Center of in vitro fertilization (IVF) between January and June 2014 were included in this prospective pilot study. The diagnosis of UEI was made with basic infertility tests. A history of CD was questioned in the initial evaluation. Anti-gliadin, anti-endomysial, and tissue transglutaminase antibodies as well as total IgA were tested. Gastroscopy was performed in patients with positive serologic tests. Histopathological CD diagnosis was made according to Marsh criteria. RESULTS: The mean age of the study population was 33.40±4.59 years. Out of the 65 couples who were included into the study group, one of the five couples was positive for the autoantibodies (7.69%). Out of these 65 couples, none of them had autoantibody positivity at the same time in both partners. Anti-gliadin antibodies were found to be positive for two females out of five couples and in three male partners of the same group. Out of these five couples, only one male partner had all the antibodies as positive (1.5%). In the histopathological examination of patients with positive autoantibodies, only the patient in whom all autoantibodies were positive had findings compatible with Marsh IIIa gluten enteropathy. Only one couple had a diagnosis of CD (1.5%). CONCLUSION: In many studies, CD was shown to affect the reproductive system of women. CD may also cause a decrease in fertility in men by affecting sperm motility and androgen levels. Our study is based on a limited sample size. Our data should be confirmed in a larger cohort of subjects. These results suggest that investigation of both couples with a diagnosis of UEI may be more beneficial in clarifying the etiology.

Prevalence of celiac disease autoimmunity in children with type 1 diabetes: regional variations across the Øresund strait between Denmark and southernmost Sweden.

OBJECTIVES: The aim was to determine the prevalence of celiac disease autoimmunity in children with type 1 diabetes (T1D) diagnosed in Denmark and Sweden. METHODS: A total of 662 Swedish children with T1D were matched with 1080 Danish children with T1D and 309 healthy children from Sweden and 283 from Denmark served as controls. Sera were analyzed for the presence of IgA and IgG (IgAG) autoantibodies against deamidated gliadin peptide (DGP) and tissue transglutaminase (tTG) with enzyme-linked immunosorbent assay (ELISA) and IgG-tTG separately in a radioligand binding assay (RBA). Human leukocyte antigen (HLA)-DQB1 and DQA1 genotyping were determined in the T1D cohorts. RESULTS: In the Swedish T1D cohort, 17.2% (114/662) were IgAG-DGP/tTG positive compared with 11.7% (126/1080) in the Danish T1D cohort (p = 0.001) and with 9.4% (29/309) Swedish (p = 0.001) and 5.7% (16/283) Danish (p = 0.003) controls. In the Swedish T1D cohort, both levels of IgAG-DGP/tTG and IgG-tTG were higher compared with the levels in the Danish T1D (p < 0.001). In the control group, 2.8% of the Danish children were positive for both IgAG-DGP/tTG and IgG-tTG, compared to 0.3% of the Swedish. Presence of HLA-DQ2 was equally distributed among 89 children with T1D positive for both IgAG-DGP/tTG and IgG-tTG. CONCLUSION: The discrepancy in levels of IgAG-DGP/tTG and IgG-tTG between Swedish and Danish T1D cohorts was independent of HLA and suggests that regional variations in comorbidity of celiac disease in T1D is caused by difference in exposure to environmental factors.

Diagnostic utility of modified gliadin peptide antibody assays in New Zealand children.

OBJECTIVE: The aim of the present study was to evaluate a panel of different antibody assays, including second-generation antigliadin kits, in a local paediatric population thought to be at risk for coeliac disease (CD). METHODS: Seventy-nine children, who tested positive for immunoglobulin A (IgA) antibodies to tissue transglutaminase (TG), underwent duodenal biopsy. At endoscopy, serum was collected from all of the patients, and 9 different coeliac antibody assays were performed, both as isolated assays and in combination. These included immunoglobulin A (IgA) anti-tissue transglutaminase (TGA), and IgA plus IgG anti-deamidated gliadin peptide (DGPAG). A diagnosis of CD was made if the biopsies showed Marsh grade 3 lesions. RESULTS: Twenty-four of 79 children had CD confirmed histologically. Only 39 of 79 were positive for Inova TGA, and 35 of 79 were positive for Inova DGPAG. Twenty-four of 39 who were TGA positive and 24 of 35 who were DGPAG positive had confirmed CD on biopsy. There was good correlation between TGA and DGPAG-positive predictive values. None of the modified gliadin tests produced false-negative results, and neither did the TGA. CONCLUSIONS: The Inova DGPAG and TGA assays have similar use in predicting CD in a selected paediatric population; however, in children who are positive for TGA when screened for CD, more than half have negative TGA serology when repeat testing is done at the time of biopsy. Those with persistent TGA positivity have only a 61.5% probability of having histologic CD, compared with 68.6% of those children positive for DGPAG.

Are transglutaminase 2 inhibitors able to reduce gliadin-induced toxicity related to celiac disease? A proof-of-concept study.

PURPOSE: Celiac disease is an autoimmune-mediated enteropathy characterized by adaptive and innate immune responses to dietary gluten in wheat, rye and barley in genetically susceptible individuals. Gluten-derived gliadin peptides are deamidated by transglutaminase 2 (TG2), leading to an immune response in the small-intestinal mucosa. TG2 inhibitors have therefore been suggested as putative drugs for celiac disease. In this proof-of-concept study we investigated whether two TG2 inhibitors, cell-impermeable R281 and cell-permeable R283, can prevent the toxic effects of gliadin in vitro and ex vivo. METHODS: Intestinal epithelial Caco-2 cells were treated with peptic-tryptic-digested gliadin (PT-gliadin) with or without TG2 inhibitors and thereafter direct toxic effects (transepithelial resistance, cytoskeletal rearrangement, junction protein expression and phoshorylation of extracellular-signal-regulated kinase 1/2) were determined. In an organ culture of celiac-patient-derived small-intestinal biopsies we measured secretion of TG2-autoantibodies into the culture medium and the densities of CD25- and interleukin (IL) 15-positive cells, forkhead box P3 (FOXP3)-positive regulatory T cells (Tregs) and Ki-67-positive proliferating crypt cells. RESULTS: Both TG2 inhibitors evinced protective effects against gliadin-induced detrimental effects in Caco-2 cells but the cell-impermeable R281 seemed slightly more potent. In addition, TG2 inhibitor R281 modified the gluten-induced increase in CD25- and IL15-positive cells, Tregs and crypt cell proliferation, but had no effect on antibody secretion in celiac-patient-derived biopsies. CONCLUSIONS: Our results suggest that TG2 inhibitors are able to reduce certain gliadin-induced effects related to responses in vitro and ex vivo.

In-vitro and in-vivo binding activity of chicken egg yolk immunoglobulin Y (IgY) against gliadin in food matrix.

Chicken egg yolk immunoglobulin Y (IgY) is a promising alternative for the prevention of enteric gliadin absorption, the predisposing factor of celiac disease (CD). IgY antibody was produced from the egg yolk of Single Comb White Leghorn chickens during the immunization period for the development of an oral immunotherapeutic agent. Here, we report the potential use of spray dried IgY antibody formulation using sugar protectants (mannitol, sorbitol, or microcrystalline cellulose powder (MCCP)). The long-term stability of the spray dried egg yolk powder formulated with 37.5% mannitol (EYP-M) preserved IgY antibody activity at 99.9%, which was significantly higher than that with other protectants (p < 0.05). In a dissolution test, the EYP-M shows 82.4% IgY activity after 2 h in simulated gastric fluid (SGF). A competitive ELISA at 50% inhibition (IC(50)) shows that 1.6 mg/mL EYP-M bound to 7.6 mg/mL and 10.5 mg/mL gliadin in SGF without and with food matrix conditions, respectively, whereas in simulated intestinal fluid, the formulation bound to 10 mg/mL gliadin, regardless of a food matrix. In-vivo study: BALB/c mice fed with EYP-M and gliadin at a ratio of 1:5 (w/w) demonstrated that gliadin absorption in the gastrointestinal tract was minimal at <1%. Thus, EYP-M containing IgY antibody may be used in CD patients to eliminate the effects of ingested toxic gliadin.

Efficacy of budesonide therapy in the early phase of treatment of adult coeliac disease patients with malabsorption: an in vivo/in vitro pilot study.

1. Budesonide is a glucocorticosteroid with a local anti-inflammatory effect. Coeliac disease is an immune-mediated disease caused by gluten ingestion in intolerant patients. The aim of the present study was to investigate the efficacy of budesonide in malabsorptive coeliac patients and its effect in an in vitro gliadin challenge. 2. Twenty coeliac patients with malabsorption were enrolled in the present study and were randomly assigned to one of two 4 week treatments: (i) a gluten-free diet alone; or (ii) a gluten-free diet plus 6 mg budesonide daily. At the end of 4 weeks treatment, all patients underwent clinical evaluation, laboratory tests and self-evaluation of well-being using a visual analogue scale. Intestinal biopsies from five coeliac patients (selected randomly) and four non-coeliac disease controls who underwent upper endoscopy for intestinal bleeding were challenged with gliadin (0.5 mg/mL) and budesonide (10-30 microg/mL) for 3 and 24 h. Biopsies were tested by immunohistochemistry and immunofluorescence for known markers of inflammation. 3. Treatment of patients with 6 mg budesonide daily for 4 weeks resulted in increased bodyweight, a decreased number of evacuations and decreased stool weight compared with patients on a gluten-free diet alone for 4 weeks. Well-being scores were higher in patients treated with both a gluten-free diet and budesonide compared with those receiving a gluten-free diet alone. 4. In vitro studies showed that budesonide reduced epithelial tyrosine phosphorylation and expression of histocompatibility leucocyte antigen complex DR (HLA-DR) elicited by gliadin-derived peptides. In addition, the expression of cyclo-oxygenase (COX)-2 and intercellular adhesion molecule (ICAM)-1 in the lamina propria was reduced in patients treated with both gliadin and budesonide compared with patients treated with gliadin alone. Budesonide alone decreased HLA-DR in crypt enterocytes, as well as ICAM-1 and COX-2 expression in the lamina propria of biopsy specimen of coeliac patients. Budesonide had no effect in control samples. 5. In conclusion, the results of the present study indicate that budesonide shows efficacy in the treatment of symptoms in adult coeliac patients with overt malabsorption. The mechanism underlying the effects of budesonide in reducing symptoms was elucidated by in vitro studies involving a gliadin challenge.

Live probiotic Bifidobacterium lactis bacteria inhibit the toxic effects induced by wheat gliadin in epithelial cell culture.

Wheat gliadin induces severe intestinal symptoms and small-bowel mucosal damage in coeliac disease patients. At present, the only effective treatment for the disease is a strict life-long gluten-free diet. In this study we investigated whether probiotics Lactobacillus fermentum or Bifidobacterium lactis can inhibit the toxic effects of gliadin in intestinal cell culture conditions. The ability of live probiotics to inhibit peptic-tryptic digested gliadin-induced damage to human colon cells Caco-2 was evaluated by measuring epithelial permeability by transepithelial resistance, actin cytoskeleton arrangements by the extent of membrane ruffling and expression of tight junctional protein ZO-1. B. lactis inhibited the gliadin-induced increase dose-dependently in epithelial permeability, higher concentrations completely abolishing the gliadin-induced decrease in transepithelial resistance. The same bacterial strain also inhibited the formation of membrane ruffles in Caco-2 cells induced by gliadin administration. Furthermore, it also protected the tight junctions of Caco-2 cells against the effects of gliadin, as evinced by the pattern of ZO-1 expression. We conclude thus that live B. lactis bacteria can counteract directly the harmful effects exerted by coeliac-toxic gliadin and would clearly warrant further studies of its potential as a novel dietary supplement in the treatment of coeliac disease.

Estudio de la enfermedad celíaca en la población pediátrica de Cantabria y sus familiares de primer grado / Study of celiac disease in the pediatric population of Cantabria (Spain) and first-degree relatives

Introducción: La enfermedad celíaca (EC) es una entidad mediada por fenómenos autoinmunes, que se presenta en sujetos susceptibles genéticamente. El 90% de los pacientes presenta el heterodímero HLA-DQ2, y el 10% restante suele presentar el HLA-DQ8. Objetivo: Estudiar las características de la EC en la población pediátrica de Cantabria y en sus familiares de primer grado, fundamentalmente en los aspectos relacionados con el haplotipo, la serología y sus formas de presentación clí nica. Pacientes y métodos: Estudio de 86 pacientes celíacos y 215 familiares de primer grado. Se recogieron datos clínicos, analíticos, inmunológicos, histológicos y de tipificación genómica. Resultados: El 95% de los caso se iniciaron con clínica clásica y el 5% eran formas monosintomáticas. Un 95% presentaba positividad a anticuerpos antigliadina (AAG) y antitransglutaminasa (AATG), y eran negativos el 5% (todos con déficit de IgA). Genotípicamente, un 71% eran portadores del DQ2 (incluidos los homocigotos y los heterocigotos), y un 9,5% del DQ8. Un 22% no presentaba heterodímero de riesgo. En el estudio familiar se hallaron 6 familiares con EC (3 AAG positivos y 4 AATG positivos). Del total, el 49% de los familiares portaba el DQ2, un 15% el DQ8, y un 40% no presentaba el heterodímero de riesgo. Conclusiones: El HLA más prevalente en nuestra comunidad fue el DQ2 (71%), claramente menor que lo publicado en nuestro medio. La prevalencia de EC en familiares de primer grado fue similar al resto de España (2,8%). Nuestros datos apoyan la necesidad del estudio sistemático en familiares de primer grado de pacientes celíacos

Background: Celiac disease (CD) is an autoimmune disease that affects genetically predisposed individuals. The HLA-DQ2 heterodimer is present in nearly 90% of patients while HLA-DQ8 is found in the remaining 10%. Aim: To study the characteristics of CD in pediatric patients in Cantabria and their first-degree relatives, with special emphasis on factors related to haplotype, serology, and forms of clinical presentation. Patients and methods: Eighty-six patients with CD and 215 first-degree relatives were HLA genotyped. Clinical, laboratory, immunologic, and histological data were obtained from all patients. Results: Clinical presentation was classical in 95% of the patients and mono-symptomatic in the remaining 5%. Anti-gliadin antibodies (AGA) and anti-transglutaminase antibodies (ATGA) were positive in 95% of the patients and negative in 5% (all with IgA deficiency). DQ2 was found in 71% of the patients (homozygotes or heterozygotes) and DQ8 was found in 9.5%. No heterodimers of risk were found in 22%. CD was found in six relatives (three were positive for AGA and four were positive for ATGA). Forty-nine percent of the relatives carried the DQ2 heterodimer and 15% the DQ8 heterodimer; no heterodimers of risk were found in 40%. Conclusions: The most prevalent HLA found in patients with CD in the autonomous region of Cantabria was DQ2 (71%). This prevalence is clearly lower than that reported in other Spanish regions. The prevalence of CD among first-degree relatives was similar to that found in other studies performed in Spain (2.8%). Our data support the need for systematic study of the first-degree relatives of patients with CD

Enfermedad celíaca / Celiac disease

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Antagonists and non-toxic variants of the dominant wheat gliadin T cell epitope in coeliac disease.

BACKGROUND: Coeliac disease (CD) is due to an inappropriate T cell mediated response to specific gluten peptides. Measured by interferon gamma (IFN-gamma) ELISPOT, about half of the gliadin specific T cells induced with in vivo wheat gluten exposure in HLA-DQ2+ CD are specific for an alpha/beta-gliadin peptide (p57-73 QE65; QLQPFPQPELPYPQPQS) that includes two overlapping T cell epitopes (PFPQPELPY and PQPELPYPQ). AIM: To define minimally substituted variants of p57-73 QE65 universally devoid of IFN-gamma stimulatory capacity but capable of antagonising IFN-gamma secretion from polyclonal T cells specific for p57-73 QE65. METHODS: Peripheral blood mononuclear cells collected from 75 HLA-DQ2+ CD patients after in vivo gluten challenge were used in overnight ELISPOT assays to screen 218 single or double substituted variants of p57-73 QE65 for cytokine stimulatory and antagonist activity. RESULTS: The region p60-71 (PFPQPELPYPQP) and especially p64-67 (PELP) was sensitive to substitution. Twelve substitutions in p64-67 stimulated no IFN-gamma ELISPOT response. Among 131 partial agonists identified, 45 produced statistically significant inhibition of IFN-gamma ELISPOT responses when cocultured in fivefold excess with p57-73 QE65 (n = 10). Four substituted variants of p57-73 QE65 were inactive by IFN-gamma ELISPOT but consistently antagonised IFN-gamma ELISPOT responses to p57-73 QE65, and also retained interleukin 10 stimulatory capacity similar to p57-73 QE65. CONCLUSIONS: Altered peptide ligands of p57-73 QE65, identified using polyclonal T cells from multiple HLA-DQ2+ CD donors, have properties in vitro that suggest that a single substitution to certain alpha/beta-gliadins could abolish their capacity to stimulate IFN-gamma from CD4 T cells and also have anti-inflammatory or protective effects in HLA-DQ2+ CD.

Sensibilidad y especificidad de los anticuerpos antigliadina y antiendomisio realizados en el Laboratorio de Inmunología del Centro Hospitalario Pereira Rossell / The sensibility and specificity of tests using antigliadin and antiendomisin antibodies in the Immunology Laboratory of the Pereira Rossell Hospital Centre

La enfermedad celíaca se presenta con una gran heterogeneidad: formas sintomáticas, silentes y latentes. La determinación de anticuerpos antigliadina y antiendomisio es útil para definir a qué pacientes realizar biopsia de intestino delgado y para el seguimiento de los celíacos ya conocidos. Estas técnicas fueron incorporadas en 1994 en el Centro Hospitalario Pereira Rossell. Objetivo: Conocer la sensibilidad y especificidad de los anticuerpos antigliadina (IgG) y antiendomisio (IgA) para el diagnóstico de enfermedad celíaca, determinados por técnica de inmunofluorescencia en la Sección de Inmunología del Laboratorio Central del Centro Hospitalario Pereira Rossell y aplicados a población hospitalaria. Se evaluaron los resultados de la serología tomando como patrón de oro la anatomía patológica de la biopsia de intestino delgado. Para el diagnóstico de enfermedad celíaca se siguieron los criterios de 1990 de la Sociedad Europea de Gastroenterología Pediátrica y Nutrición. Se evaluó la serología de 65 niños que recibían gluten al momento de la extracción de sangre: 50 celíacos y 15 no celíacos (con biopsia normal). La sensibilidad y la especificidad fueron de 94 por ciento y 80 por ciento para los anticuerpos antigliadina y de 94 por ciento y 93 por ciento para los antiendomisio respectivamente. Estos resultados son similares a los comunicados por los mejores laboratorios a nivel internacional.

Significado y valor de los marcadores serológicos de la enfermedad celíaca / Significance and value of serological markers in celiac disease

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Anti-inflammatory effects of alpha-melanocyte-stimulating hormone in celiac intestinal mucosa.

OBJECTIVES: The peptide alpha-melanocyte-stimulating hormone (alpha-MSH) possesses potent anti-inflammatory activities and has been previously implicated in the endogenous control of inflammatory reactions. The aim of the present research was to determine whether alpha-MSH and its receptors participate in a localized anti-inflammatory response in the duodenal mucosa of celiac patients. METHODS: Three series of experiments were performed, using duodenal biopsy pairs from 53 adult celiac patients and 14 normal subjects, in order to determine: (1). mucosal immunoreactivity for alpha-MSH and melanocortin receptors (MCRs), and gene expression of alpha-MSH precursor pro-opiomelanocortin and MCRs; (2). alpha-MSH and inflammatory cytokine production by duodenal specimens in vitro, and the influence of synthetic alpha-MSH on such cytokine production, and (3). the influence of stimulation with gliadin (the subfraction of gluten that is toxic to patients with celiac disease) on alpha-MSH and cytokine production in vitro and the effect of alpha-MSH on gliadin-stimulated cytokine production. RESULTS: Elements of a localized anti-inflammatory influence based on alpha-MSH and its receptors were found: duodenal mucosa showed immunostaining for alpha-MSH and two of its receptor subtypes, MC1R and MC5R. alpha-MSH and MC1R immunoreactivity was more intense in specimens from celiac patients. Release of interleukin 6 from gliadin-stimulated duodenal mucosa was inhibited by synthetic alpha-MSH in vitro. CONCLUSIONS: Presence of alpha-MSH and its receptors in celiac mucosa suggests the presence of a local reaction to control the inflammatory response elicited by gliadin. In selected cases of refractory celiac disease, treatment with exogenous peptides might be considered.

Crecimiento en la enfermedad celiaca / Effects of celiac disease on growth

Introducción. La mala absorción crónica, como ocurre en la enfermedad celiaca (EC), conduce a una disminución del aporte de nutrientes y/o a un incremento de su pérdida que afecta al proceso de crecimiento. Uno de estos nutrientes que se encuentra en límites bajos y que está implicado en esta situación es el cinc, el cual se admite que interviene a través de la regulación de la síntesis de IGF-I. Material y métodos. Se han estudiado los niveles de Zn, Fe, IGF-I e IGFBP-3 en un grupo de 26 niños con celiaquía sometidos a dieta sin gluten durante un periodo que oscilaba entre 1 y 36 meses. Resultados. La velocidad de crecimiento -(VC)- en los niños sin gluten durante 1=3 meses fue de -2,21 ñ 0,54 DE y ascendió hasta 1,38 ñ 1,43 DE en los que llevaban entre 12 y 24 meses sin gluten, para bajar hasta 0,92 ñ 1,8 DE entre los 24 y 36 meses. Encontramos diferencias significativas entre un grupo control y los celiacos exentos de gluten durante 1-24 meses para IGF-I (p <0,01) y Zn (p <0,05).En el grupo control se estableció una correlación significativa (p <0,01) entre IGF-I e IGFBP-3 y entre estos dos parámetros y la edad ósea; correlación que se pierde en los celiacos. Por último, se halló una correlación positiva entre los niveles de Zn en los celiacos en función del tiempo sin gluten con un nivel de significación de p <0,05.Conclusión. Estos datos concuerdan con la idea de que este nutriente está relacionado con la respuesta del crecimiento en situaciones de mala absorción mediante su papel en la regulación de IGF-I (AU)

Estudio de concordancia entre anticuerpos IgA antitransglutaminasa tisular recombinante humana y anticuerpos IgA antiendomisio / Study of the agreement between Iga human recombinant tissular transglutaminase antibodies and IgA endomysial antibodies

Introducción: La enfermedad celíaca (EC) o enteropatía sensible al gluten es un trastorno intestinal de, etiología multifactorial. En su patogenia se encuentran implicados ge-nes HLA y no-HLA, el gluten dietético (trigo, avena, cebada y centeno) y posiblemente factores medioambientales adicionales. La EC se caracteriza por la presencia de atrofia vellositaria en el intestino delgado, demos-trada mediante biopsia. La prevalencia es del 0,1 por ciento. Suele producir sintomatología clí-nica entre el segundo y el tercer año de vida. La clínica clásica incluye diarrea crónica, distensión abdominal y escaso desarrollo pondostatural, sin embargo, puede aparecer mas tardíamente con una clínica más larvada, como anemia, fatiga, pérdida de peso, diarrea, estreñimiento e incluso sintomatología neurológica. El tratamiento consiste en la exclusión del gluten de la dieta. La persistencia de la exposición al gluten dietético se asocia con un riesgo elevado de anemia, infertilidad, osteoporosis y linfoma intestinal, así como aumento de la mortalidad.Para el diagnóstico de EC se utilizaron los criterios de la Sociedad Europea de Gastroenterología y Nutrición (ESPGAN). El estudio de diversos anticuerpos es de gran ayuda para el diagnóstico y seguimiento de la EC: a) los anticuerpos antigliadina de clase IgA (AGA-IgA) detectados mediante técnica ELISA presentan una sensibilidad (S) del 85 por ciento y una especifidad (E) del 82 por ciento y los de clase IgG (AGA IgG) presentan una S del 69 por ciento, y una E del 73 por ciento, y b) los anticuerpos antiendomisio de clase lg A (EMA-IgA) medidos mediante técnica de inmunofluorescencia indirecta (IIF) presentan una S del 93 por ciento y una E del 99 por ciento; y antirreticulina de tipo IgA (ARA-IgA), presentan una S del 92 por ciento y una E del 97 por ciento. Recientemente, Dieterich et al, utilizando un método de inmunoprecipitación, descubrieron un anticuerpo frente a la transglutaminasa tisular (TGt) de cobaya de tipo IgA (anti-TGt-IgA), presente en los pacientes con EC, con una S del 95 por ciento y una E del 94 por ciento. En este estudio hemos testado un nuevo anticuerpo frente a TGt recombinante humana de clase IgA (anti T GtR-IgA), medido mediante técnica de ELISA, y lo hemos comparado con la detección de anti-TGt-IgA (método utilizado actualmente) y con la detección de EMA-IgA (técnica de referencia).Materiales y métodos: Se estudiaron 133 sueros de pacientes con sospecha clínica de EC, utilizándose técnicas de IFI para detectar EMA-IgA y de ELISA para medir anti-TGt-IgA, poniéndose en marcha una nueva técnica de ELISA que detecta anti-TGtR-IgA. Resultados: Primero se compararon los anticuerpos anti-TGt-IgA en uso, respecto a los EMA-IgA, y se observó una S del 96,2010 y una E del 91,5 por ciento; valor predictivo positivo (VPP) del 74,2 por ciento y valor predictivo negativo (VPN) del 98,9 por ciento (X2= 85,56; p 4,001; alfa <0,001). Posteriormente se analizaron los mismos sueros comparando la positividad para anti-TGtR-IgA respecto a anti-TGt-IgA. Estos datos arrojan a su vez los siguientes valores: S= 74,2 por ciento; E= 98,9 por ciento; VPP= 96,2 por ciento; VPN= 91,5 por ciento (X2= 85,56; p 4,001; alfa <0,001). Por último, se comparó la positividad para anti-TGtR-IgA: respecto a EMA IgA, y se encontró una concordancia total entre ambas determinaciones: S= 100 por ciento; E= 100 por ciento; VPP= 100 por ciento; VPN= 100'0 (x2= 133; p 4,001; alfa 4,001).Conclusiones: La detección de anticuer-pos anti-TGtR-IgA presenta una sensibilidad y especifidad superiores a la detección de anti-TGt-IgA. Hemos observado una identidad total entre anti-TGtR-IgA y EMA-IgA, por tanto, la detección de anti-TGtR-IgA constituye una técnica sensible y específica que podría sustituir a la detección mediante IFI de EMA-IgA en el diagnóstico de EC. Como la detección de anti-TGtR-IgA tiene las ventajas de ser un ensayo cuantitativo, de rápida realización, no sometido a la varia-bilidad del observador y fácilmente reproducible, podría ser una herramienta muy útil en el diagnóstico y seguimiento de la EC (AU)

Relación de los anticuerpos IgA anti-transglutaminasa tisular con la situación morfológica de la mucosa intestinal en niños con enfermedad celíaca / Relation of anti-tissue transglutaminase IgA antibodies with the morphology of the intestinal mucosa in children with celiac disease

Objetivo: Se ha investigado si la presencia de anticuerpos IgA antitransglutaminasa tisular es un indicador de la existencia de lesión de la mucosa intestinal, tanto en pacientes celíacos, en sus distintas fases evolutivas, como en controles de diverso tipo.Métodos: En 260 muestras de 214 pacientes, elegidos aleatoriamente, se han determinado, por método ELISA, anticuerpos IgA antitransglutaminasa tisular utilizando transglutaminasa de hígado de cobaya, así como anticuerpos IgA e IgG anti-gliadina. En un subgrupo de 109 muestras se determinaron, además, anticuerpos IgA antitransglutaminasa con transglutaminasa humana recombinante. Los resultados de todos ellos se han comparado con los de la biopsia intestinal tomada en los mismos días.Resultados: Con transglutaminasa de cobaya se ha obtenido una sensibilidad de 0,92 y una especificidad de 0,97, con relación a la biopsia intestinal, con una validez total máxima de 0,90 que corresponde a un coeficiente de probabilidad positivo de 34. El empleo de transglutaminasa humana recombinante no mejora estos resultados, pero se obtienen menos valores dudosos. Los anticuerpos antigliadina, tanto de clase IgA como de clase IgG sólo han mostrado una sensibilidad de 0,86 y 0,42 y una especificidad de 0,77 y 0,76, respectivamente.Conclusiones: La determinación de anticuerpos IgA anti-transglutaminasa tiene gran capacidad diagnóstica, reflejando el estado de la mucosa intestinal y mejorando sensiblemente la de otros marcadores séricos. No obstante, su discriminación no es absoluta, por lo que la BI debe seguir siendo el patrón-oro de referencia para el diagnóstico definitivo de la enfermedad celíaca (AU)