EOSINOPHIL DERIVED MAJOR BASIC PROTEIN IMPACT ON INTESTINAL BARRIER FUNCTION
Glenn T Furuta, Steven J Ackerman, Edward E Nieuwenhuis, Richard Blumberg, Jamie Lee, Gerald J Gleich, Sean P Colgan, Combined Program of Pediatric Gastroenterology and Nutrition, Harvard Medical School, Boston, MA; Biochemistry, University of Illinois at Chicago, Chicago, IL; Gastroenterology, Brigham and Women's Hospital, Boston, MA; Gastroenterology, Brigham and Women's Hospital, Boston, MA; Biochemistry and Molecular Biology, Mayo Clinic, Scottsdale, AZ; Dermatology, University of Utah, Salt Lake City, MA; Anesthesia, Brigham and Women's Hospital, Boston, MA
Eosinophils lie juxtaposed to the epithelium in diseases associated with altered intestinal barrier function. We previously showed that co-culture of an eosinophil cell line with intestinal epithelia elicits a fall in barrier function.
To identify eosinophil mediator(s) which alter intestinal barrier function.
Eosinophil myelocytes (AML14.3D10) were co-cultured with T84 epithelial cells in a non-contact Transwell model. Supernatants were harvested and assessed for bioactive products in add back experiments and western blots. The impact of the eosinophil granule proteins, major basic protein (MBP) and eosinophil derived neurotoxin (EDN), on intestinal barrier was assessed by measurements of transepithelial resistance (TER) and gene specific mRNA from T84 cells. Oxazolone colitis, a hapten mediated Th2 type colitis, was induced in MBP null and eosinophil peroxidase (EPO) null mice (129/SvJ background) and wild type (WT) mice. Intestinal permeability was assessed with an orally gavaged 4kda FITC labeled probe.
MBP and EDN were identified in supernatants from eosinophil-epithelial co-cultures by Western blot analysis. Exposure of the basolateral surface of T84 cells to MBP (10-6M) led to a time and concentration dependent fall in TER (84% decline from baseline) that was maximal at 24 hrs. In contrast, TER did not change when exposed to EDN (10-6M). Occludin expression declined in T84 cells exposed to MBP (10-6M) as determined by RT-PCR. Compared to EPO null and WT mice, MBP null mice were significantly protected from weight loss following induction of colitis at day 1 [percent (%) weight loss from baseline; 7.0+/−1.1 and 6.4+/−0.8 vs. 2.2+/−0.8, EPO null and WT vs. MBP null respectively p<0.01]. MBP null mice were significantly protected from colonic shortening at day 7 compared to WT mice [colonic length (cm) at day 7; 9.6+/−0.8 vs. 7.5+/−0.9 cm MBP null vs. WT respectively p<0.05].
MBP induces alterations in intestinal barrier dysfunction and downregulates the junctional molecule occludin. MBP may be pathological mediator for inflammation observed in a T cell dependent model of colitis.
EOSINOPHILIA OF THE COLON AND ABDOMINAL PAIN IN CHILDREN
Mayssa Zayat, Mohamad S Miqdady, Carlos H Lifschitz, Department of Pediatrics, Texas Children's Hospital, Houston, TX
Chronic and recurrent abdominal pain is responsible for 2–4 % of pediatric office visits. In the majority of the cases the etiology of the pain is not clear and is thought to be a functional bowel disorder. Eosinophilia of the colon has been associated in infancy with dietary protein intolerance but it has not been investigated as much in older children. We performed a retrospective chart review of 16 patients who presented between April 1999 and October 2000 and had abdominal pain and eosinophilic colitis.
All patients had more than 20 eosinophils/ HPF. The age range was 7 to 17 yr. Six (37.5 %) were females. The duration of symptoms prior to colonoscopy ranged between 2 weeks to 2 years. The main differences with patients with irritable bowel syndrome is that those with eosinophilic colitis recalled fairly precisely when their symptoms had begun, pain occurred both during the week as well as weekends, vacation or even while playing and was not limited to the periumbilical area. None of the patients had experienced fever or joint pain, CBC was normal without peripheral eosinophilia, and erythrocyte sedimentation rate was normal in the 7 of the 16 patients in which it was obtained. Seven (44%) patients underwent an upper endoscopy as well, and mild or no increase in the number of eosinophils in the esophagus or stomach was found. Empirical treatment with a short course of low dose corticosteroids resulted in resolution of symptoms in five patients, two patients stopped treatment because of side effects, and one patient developed Crohn disease. Four patients did not improve. Four patients could not be contacted for follow up.
The association of eosinophilia of the colon and abdominal pain in children warrants further investigation.
EOSINOPHILIA AND GEOHELMINTHS IN CHILDREN UNDER TWELVE YEARS
Carlos A Velasco, Doris P Lopez, Zoraida Tarazona, Leonor Sanabria, Carmen H Sanabria, Isabel C Sarmiento, Eliana Rodriguez, Gastroenterologia y Nutricion Pediatrica, Universidad del Valle, Cali, Valle, Colombia; Facultad de Salud, Universidad Industrial de Santander, Bucaramanga, Santander, Colombia
The prevalence of intestinal parasitism in Bucaramanga, Colombia is of 86%, being described eosinophilia (eosinophiles > 500/mm3) mainly for geohelminths (GH) as A. lumbricoides, S. stercoralis, hookworms and T. trichiura.
To determine the association between eosinophilia (E) and GH in children younger than 12 years.
Patient and methods:
A prospective descriptive study, transversal observational carried out in children under 12 years hospitalized during a period of a year in the HURGV of Bucaramanga, Colombia. 3 ml of blood were obtained for the absolute recount of eosinophiles and 3 stool analysis for the search of GH in patients without allergic antecedents, medications causing E, antibiotic against parasites and without systemic illnesses. Data like age, gender, weight and height were obtained. t student test, and X2, were used being a p < 0.05 significant.
130 children were included, with ages between 1 month and 12 years (41±36 months, 70 masculine), 87% from Bucaramanga and 51% with malnutrition. 32 children presented GH: 10 with ascaris, 7 with hookworms, 5 with strongyloides, 4 with whipworm infection and 6 with more than one GH; 35 children evidenced E: 16 mild (between 500 and 1000 eosinophiles/mm3), 7 moderate (between 1000 and 1500 eosinophiles/mm3) and 12 severe (> 1500 eosinophiles/mm3). When analyzing the 32 children with GH a significant association with E was observed ( p = 0.015, 95% CI, OR = 4.74), however, when this association was isolated for each GH, no association was observed ( p = 0.1719 ), neither when it was correlated for E grades ( p > 0.05).
In our population, we found a positive association between GH and E; however, this association was not identified when it was analysed for each GH, being the prevalence for GH of 25%.
PANCREATIC ENZYME SUPPLEMENTS IMPROVE SEVERE ECZEMA IN CHILDREN WITH MULTIPLE FOOD ALLERGIES
Sanford Singer, Jonathan Meddings, Julie Powell, Anne Desroches, Ernest Seidman, Divisions of GI & Nutrition, Dermatology and Allergy, Sainte Justine Hospital, University of Montreal, Montreal, QC, Canada; Division of Gastroenterology, Dept. of Medicine, University of Calgary, Calgary, AB, Canada
A basis of the allergic response to a food is uptake of allergen(s) across a hyperpermeable intestinal mucosa. We hypothesized that accelerated hydrolysis of dietary allergens in vivo using pancreatic enzyme supplements would decrease allergen uptake, resulting in improvement of severe eczema.
Pediatric patients (n=10) with severe eczema and multiple food allergies resistant to conventional therapies (dietary, topical) were recruited for study. Baseline atopic dermatitis severity was measured using the SCORAD index. Intestinal permeability was analyzed by HPLC measurement of sucrose, lactulose and mannitol excretion in urine after a test dose. Patients then received pancreatic enzymes (Creon) without other changes in therapies for 6 weeks. SCORAD and permeability measures were then repeated.
Pancreatic enzyme supplements significantly improved severe eczema in 80% of the patients studied. Although small bowel permeability, as measured by urinary LAC/MAN testing was slightly greater in children with allergies compared to controls, the difference was not statistically different, likely due to the small numbers of subjects. Further placebo controlled trials are warranted.
Supported by a research grant from Solvay Pharma.
HYPEREOSINOPHILIA: A SIGN OF MILK ALLERGY IN THE PREMATURE INFANT?
Seung Dae Park, James F Markowitz, Anupama Chawla, Fredric Daum, Pediatrics, Schneider Children's Hospital at North Shore, Manhasset, NY
While eosinophilia (absolute eosinophil count (AEC) >700/mm2) is common in premature infants, marked elevations (AEC>1400/mm2) are unusual. It has been suggested that eosinophilia is associated with RBC transfusions, TPN, bronchopulmonary dysplasia, bacterial infection, necrotizing enterocolitis, and use of erythropoietin. Frequently, however eosinophilia is ignored as a non-specific finding. We have recently identified a series of premature babies with feeding intolerance and marked hypereosinophilia in whom introduction of amino acid-based formula promptly resulted in improvement of clinical symptoms and reduction in AEC.
Four premature infants were evaluated at the NICU between July, 2001 and April, 2002. Charts were reviewed retrospectively. These infants were referred for signs and symptoms including abdominal distension, apnea ± bradycardia with feeding, vomiting, hematochezia, diarrhea, or edema with hypoalbuminemia. All had markedly elevated AEC (See Table) which had previously been discounted as a non-specific finding by the attending neonatologists. Three of these four infants were already on casein hydrolysate formula as treatment for abdominal distension, apnea ± bradycardia with feeding, or hematochezia. Formula was switched to casein hydrolysate formula for the one infant who was on cow milk protein-based formula, and to amino acid-based formula for those already receiving casein hydrolysate. Hypereosinophilia and clinical symptoms improved by 2 – 17 days after formula was switched or withheld (See Table). The infant (Pt. 4) whose formula was switched to casein hydrolysate initially improved, but ultimately developed recurrent hypereosinophilia which resolved within 8 days following subsequent introduction of amino acid-based formula (See Table).
Marked hypereosinophilia in premature infants with feeding problems should not be ignored as a non-specific finding. Milk protein or casein hydrolysate allergy can cause marked increase in AEC in premature infants. The use of an amino acid-based formula rapidly results in clinical improvement and reduction of AEC.