Salvatore Accomando

Hsp60 and Hsp10 increase in colon mucosa of Crohn’s disease and ulcerative colitis

The purpose of this work was to determine in colon mucosa of Crohn’s disease (CD) and ulcerative colitis (UC) in relapse: a) the levels of the chaperonins Hsp60 and Hsp10; b) the quantity of inflammatory cells; and c) if the levels of chaperonins parallel those of inflammation cells. Twenty cases of CD and UC and twenty normal controls (NC) were studied using immunohistochemistry, Western blotting and immunofluorescence. Immunohistochemically, Hsp60 and Hsp10 were increased in both inflammatory bowel diseases (IBD) compared to NC. These results were confirmed by Western blotting. Hsp60 and Hsp10 occurred in the cytoplasm of epithelial cells in CD and UC but not in NC. Hsp60 and Hsp10 co-localised to epithelial cells of mucosal glands but not always in connective tissue cells of lamina propria, where only Hsp60 or, less often, Hsp10 was found. Cells typical of inflammation were significantly more abundant in CD and UC than in NC. Since chaperonins are key factors in the activation of the immune system leading to inflammation, we propose that they play a central role in the pathogenesis of the two diseases, which, consequently, ought to be studied as chaperonopathies.

Are food intolerances and allergies increasing in immigrant children coming from developing countries

There are not available data concerning the occurrence, the clinical features and the environmental risk factors for food intolerances and allergies in immigrant children. The aim of the study was to evaluate rates, distribution, clinical features and environmental risk factors for food intolerances and allergies in immigrant children. Hospital records of 4130 patients with celiac disease (CD), cow milk protein intolerance (CMPI) and food allergies (FA) diagnosed in 24 Italian Centres from 1999 to 2001 were retrospectively reviewed, comparing immigrant patients with Italian ones. 78/4130 (1.9%) patients were immigrant: 36/1917 (1.9%) had CD, 24/1370 (1.75%) CMPI and 18/843 (2.1%) FA. They were evenly distributed across Italy and their native areas were: East Europe (23/78), Northern Africa (23/78), Southern Asia (14/78), Saharan and Sub‐Saharan Africa (9/78), Southern America (4/78), Far East (3/7), Middle East (2/78). Despite differences in their origin, the clinical features of immigrant children were similar to the ones of Italian patients and among each ethnic group. The majority of them were born in Italy (57/78) or have been residing in Italy since several years (19/78). All of them had lost dietary habits of the native countries and had acquired those of the Italian childhood population. Food intolerances and allergies are present also in children coming from developing countries, and paediatricians will need to have a full awareness of them because the number of immigrant children in Italy is quickly increasing. The clinical features of food intolerances and allergies appear the same in each ethnic group, despite differences in races. Sharing of dietary habits with the Italian childhood population seems to be an important environmental risk factor.

Changes in immunohistochemical levels and subcellular localization after therapy and correlation and colocalization with CD68 suggest a pathogenetic role of Hsp60 in ulcerative colitis.

In an earlier work, the role of heat shock protein (Hsp60) in the pathogenesis of ulcerative colitis (UC) was suggested by its significant increase in the pathological mucosa parallel with an increase in inflammatory cells. More data in this direction are reported in this work. We analyzed by immunohistochemistry biopsies of colon tissue from 2 groups of patients with UC and treated with either 5-aminosalicylic acid (5-ASA) alone or in combination with a probiotic. We looked for inflammatory markers and Hsp60. Both the treatments were effective in reducing symptoms but the group treated with both 5-ASA and probiotics showed better clinical results. Amelioration of symptoms was associated with reduction of both inflammation and Hsp60, a reduction that was most marked in the group treated with 5-ASA and probiotics. The levels of Hsp60 positively correlated with those of CD68-positive cells, and double immunofluorescence showed a high index of colocalization of the chaperonin and CD68 in lamina propria. Immunoelectron microscopy showed that Hsp60—classically a mitochondrial protein—was abundantly also present in cytosol in biopsies taken at the time of diagnosis, but not after the treatment. Our data suggest that Hsp60 is an active player in pathogenesis of UC and it can be hypothesized that the chaperonin is responsible, at least in part, for initiation and maintenance of disease.

Infliximab administration effective in the treatment of refractory Kawasaki Disease

Dear Sirs, Kawasaki Disease (KD) is a common vasculitis during the pediatric age. It involves little and medium caliber arteries. A risk of poor outcome is linked to the development of coronary artery aneurysms with sudden thrombo-embolic evolution (1). KD can present either in a classical (fulfilling all the American Reumatism Association (ARA) criteria) or an atypical or an incomplete form. Nowadays, the mainstay therapy for KD consists in high doses of IV administration of Immunoglobulins. Refractory cases, which do not respond to this kind of therapy, are demonstrated (2). Infliximab is a chimerical monoclonal antibody (IgG1) against Tumor Necrosis Factor (TNF) alpha. It belongs to biologic drugs, and it is properly used in the management of IBD, for adult patients in particular. We present a typical case of KD occurred in a 11 months old male child, who did not react both to IVIg administration and methyl prednisolone pulse therapy. A.A. was admitted after 4 days of continuous fever resistant to antibiotic therapy. Cutaneous rash, cheilitis, and conjunctivitis appeared after the fifth day of fever, and neck lymphadenopathy was also present. Phlogistic indexes were high: CrP (12 mg/dl), platelets (650,000/mm). Leucocitosis (WBC 22,000/mm) was present to total blood cell count. The first ultrasound cardiac evaluation showed no specific signs of coronaritis. Ultrasound abdominal evaluation revealed spread gall bladder and a target image of the bowel. A 2 gr/Kg intravenous Immunoglobulins administration was started, together with administration of salicylic acid at 80 mg/kg. The patient did not respond to therapy and fever persisted. A second dose of Ig was administered at the same amount. Ultrasound cardiac evaluation, performed after the first week of disease, showed two light coronary aneurisms. Patient s general conditions got worse. Hepatic serum transaminases also increased (three times normal values). So,methyl prednisolone iv pulse at 30 mg/ Kg was performed for three consecutive days. Fever persisted, CrP (18 mg/dl), platelets (1,400,000/mm), and white blood cells (45,000/ mm) raised. A second cardiac evaluation showed the patient was worsening. As a rescue therapy, Infliximab was started at 5 mg/Kg (3). The total infusion took 6 h and no adverse reaction occurred. The clinical response to Infliximab was excellent. Fever sharply decreased. Twenty-four h after the infusion, CrP (8 mg/dl), white blood cells (21,000), and hepatic serum transaminases (one time and half normal values) strongly decreased. Rash, cheilitis, and conjunctivitis disappeared. Ten days after the infusion, clinical remission persisted and finger desquamation was present to both hands and feet. CrP (0.4 mg/dl), white blood cells, and hepatic serum transaminases reached normal values. The patient was discharged after last cardiac ultrasound evaluation, which expressed a stopped disease, and he entered a strict surveillance follow-up program, ongoing the assumption of salicylic acid at 5 mg/ Kg as antiplatelets aggregation drug. Our report describes the use of Infliximab in the youngest patient affected by refractory KD as long as we know. KD is a reactive inflammatory condition, probably in genetic susceptible individuals (4), and TNF alpha and other pro-inflammatory cytokines may play an active role in its pathogenesis. In particular, patients with refractory KD, who are at significant risk for thromboembolic events because of their higher TNF alpha levels, may also benefit by Infliximab therapy, thanks to the new properties of the biologic drug recently reported by Danese et al. (5) and by Di Sabatino et al. (6). Indeed, Infliximab acts reducing the molecular expression of intestinal endothelial molecules vascular cell Pediatr Allergy Immunol 2010: 21: 1091–1092

A propensity score-matched comparison of infliximab and adalimumab in TNF-α inhibitors naïve and non-naïve patients with Crohn’s disease: real-life data from the Sicilian Network for Inflammatory Bowel Disease (SN-IBD)

Background and Aims There is an unmet need to better understand the effectiveness of different biologics in inflammatory bowel diseases. We aimed at performing a multicentre, real-life comparison of the effectiveness of infliximab [IFX] and adalimumab [ADA] in Crohns disease [CD]. Methods Data of consecutive patients with CD treated with IFX and ADA from January 2013 to May 2017 were extracted from the cohort of the Sicilian Network for Inflammatory Bowel Disease. We used propensity score-matching accounting for the main baseline characteristics in TNF-α inhibitor-naïve and non-naïve patients. Results A total of 632 patients [735 total treatments] were included. Among naïve patients, a clinical benefit [the sum of steroid-free remission plus clinical response] was achieved in 81.8% patients treated with ADA and in 77.6% patients treated with IFX (adjusted odds ratio [OR]: 1.23, 95% CI 0.63-2-44, p = 0.547] at 12 weeks; after 1 year, a clinical benefit was achieved in 69.2% of patients treated with ADA and in 64.5% patients treated with IFX [adjusted OR: 1.10, 95% CI 0.61-1.96, p = 0.766]. Among non-naïve patients, a clinical benefit was achieved in 61.7% of patients treated with ADA and in 68.1% of patients treated with IFX [adjusted OR: 0.72, 95% CI 0.21-2.44, p = 0.600] at 12 weeks; after 1 year, a clinical benefit was achieved in 48.9% of patients treated with ADA and in 40.4% patients treated with IFX [adjusted OR: 1.23, 95% CI 0.54-2.86, p = 0.620]. Conclusions In this propensity score-matched comparison of ADA and IFX in CD, both drugs showed high rates of clinical benefit, without significant differences between them.

PO-0994 Ltri In Paediatrics: Analysis Of An Annual Survey

Background and aims Lower tract respiratory infections (LTRI) are very frequent in paediatric population. We collected epidemiological, etiological and clinical data and correlated them to some variables. Methods We performed an observational study of all children with a LTRI, admitted to our Paediatric Unit from February 2013 to January 2014. The population was grouped in three classes of ages (0–2yr, 3–6yr, >6yr). All patients underwent to a questionnaire focused on exposition to protective and risk factors for respiratory diseases. Results 83 children were included. We grouped them according to discharge diagnosis and analysed the distribution for ages, sex and season of onset. At the admission 18 patients had respiratory distress; O2 therapy was necessary for 11 of them, infusion therapy for 40, endovenous antibiotic therapy for 34. Swabs were positive in 1 case for Parainfluenza viruses and in 2 for S. Aureus; sierological tests were positive in 5 cases for Mycoplasma Pneumoniae, in 2 for Chlamydia Pneumoniae, in 1 for ParvovirusB19, in 1 for Coxsackievirus. In 28 patients (33%) exposition to passive smoke was observed, in 17 (20%) to aeroallergens and in 33 (40%) a personal or familiar story of atopy. Conclusions Our experience showed a higher prevalence of LTRI in males (63%), in winter (35%) and spring (33%), without a difference between preschool and school age children.

Increased percentages of calprotectin and TNF-Α double-positive monocytes in the acute phase of Kawasaki disease

Background The acute phase of KD is characterized by a deficiency of suppressor T cells, marked activation of the immune system and increased secretion of cytokines by immune effector cells. Moreover, it has been shown that myeloid-related protein (MRP-8 and MRP-14) and S100proteins, the major calcium-binding proteins secreted by activated neutrophils and monocytes, contribute to cause inflammation in acute lesions of KD, and indeed one of the more common hematological alteration in KD is the increase of peripheral blood monocytes. Calprotectin, one of the major calcium-binding proteins, can lead to direct and indirect effects that result not only in inflammation but also in modification of microvascular wall in acute vasculitis syndromes.

PA83 DOUBLE BLIND PLACEBO CONTROLLED FOOD CHALLENGE USEFUL TO DISCONFIRM OVER ESTIMATED DIAGNOSIS OF CMPA IN CHILDREN

Methods: 14 patients, (12 months-12 yrs) previously diagnosed as having CMPA, underwent our diagnostic algorithm in order to confirm or to exclude diagnosis. Diagnostic algorithm includes: total blood cell count, serum IgE assay, RAST, betalactotest, Prick by prick with fresh milk, chemical examination and eosinophilic cell count of the stools. DBPCFC was performed with extensively hydrolyzed formula (as placebo) VS a lactose-free, cow milk derived formula.

OC.01.3 ROLE OF NAKED CUTICLE HOMOLOG 1 GENE ON CHR 16Q12 IN INFLAMMATORY BOWEL DISEASE

In both main forms of Inflammatory bowel disease (IBD), ulcerative colitis (UC) and Crohns disease (CD), intestinal commensal microbiota are thought to trigger the disease in genetically susceptible individuals. Approximately one third of patients with CD have a mutation in the NOD2 gene that encodes an intracellular bacterial pattern-recognition receptor located within Paneth-cells. The maturation of Paneth cells is regulated via the canonical Wnt cascade. The Nkd1 (naked cuticle homologue 1), gene, a Wnt antagonist, is located near NOD2 at 16q12, and it has been found weakly associated with CD in WTCCC dataset (rs8047222 P<1x10-5). Aims: To investigate rs4785433, rs730055, and rs8047222 SNPs of NKD1 and to search for likely correlation with clinical phenotypes in both adult and pediatric onset IBD patients. Methods: 1185 CD patients (670 male) with a mean age at diagnosis of 27 yrs (range 1-80) (334 < 19 yrs at diagnosis), 1385 UC patients (803 male) with a mean age at diagnosis of 31 yrs (range 1-83) (246 < 19 yrs at diagnosis), and 645 healthy controls (HC), were genotyped for NKD1 gene by using the TaqMan allelic discriminationmethod. Genotype frequencies were comparedwith respect to age at diagnosis, disease localization and behaviour (Montreal classification), need of resective surgery, response to medical therapy (steroids, immunosuppressive drugs), and NOD2 status. Results: A significant allele and genotype association of the rs8047222 with UC (P=0.01, OR=0.83, CI=0.73-0.96; P=0.02, OR=0.79; CI=0.65-0.96) was shown, especially in adult-onset cohort (P<0.002). After stratifying the cohort on the basis of NOD2 genotypes (NOD2-=no major variants; NOD2+=at last one variant), a significant genotype association in NOD2patients (P=0.004; OR= 0.74; CI=0.61-0.91) was also demonstrated. No significant difference for either allele and genotype frequencies with rs8047222 was found for CD compared with controls, however, a significant genotype association in NOD2patients (P=0.03, OR=0.77, CI=0.62-0.97) was found. At genotype-phenotype evaluation, a reduced freq uency of distal (E1) vs extensive (E3) colitis in UC (47% vs 78%; P=0.006; OR=0.25, CI=0.09-0.70), and colon (56%) vs ileum localization in CD(65%)(P=0.02, OR=0.69, CI=0.49-0.95) was demonstrated. No significant difference for either allele and genotype frequencies for rs4785433, rs730055 SNPs for IBD was found. Conclusions: The rs8047222 SNP on NKD1 gene has a significant protective role in both UC and CD, possibly influenced by NOD2 status, with interaction with disease localization. Nkd1 might be responsible for the residual linkage at 16q12 locus in NOD2-negative CD patients.