Abstract no.: P3.5 DIFFERENTIAL EXPRESSION OF HUMAN BETA DEFENSIN- 2 AND -3 IN GASTRIC MUCOSA OF HELICOBACTER PYLORI-INFECTED INDIVIDUALS B. Bauer,* T. Wex,   D. Kuester,   T. Meyer* and P. Malfertheiner   *Max Planck Institute for Infection Biology, Berlin, Germany;   Otto-von-Guericke University Magdeburg, Magdeburg, Germany Background: Antimicrobial peptides are keyplayers of initial innate immune responses to human pathogens. Two major representatives, the human beta de- fensin 2 and 3 (hBD2, hBD3) are both known to be induced by Helicobacter pylori. Previously, it was demonstrated in vitro that H. pylori actively abrogates hBD3 expression during prolonged infections. Here we comprehensively assessed hBD2 and hBD3 expression ex vivo in the gastric mucosa of healthy individuals. Materials and Methods: Twenty volunteers (H. pylori positive and H. pylori negative: n = 10) were enrolled. H. pylori positive subjects underwent eradication therapy and repeated the protocol. Expression of both defensins were assessed by quantitative RT-PCR and ELISA, and correlated with histopathological degree of gastritis. Results: hBD2 and hBD3 were found to be ubiquitously expressed in all three groups. In general, hBD2 levels were elevated in relation to H. pylori infection (up to 40-fold). This upregulation correlated with degree of gastritis in corpus and antrum. In contrast, hBD3 mRNA amounts were significantly decreased, while corresponding protein levels remained unchanged. Eradication therapy led to normalization of mucosal hBD2 expression, while hBD3 expression demonstrated high interindividual variations among individuals. Conclusions: Both defensins are ubiquitously but differentially expressed in gastric mucosa in relation to H. pylori infection. Ex vivo data support previous in vitro findings that H. pylori infection is associated with reduced hBD3 expression in chronic active gastritis. Abstract no.: P3.6 BLOOD AND LYMPHATIC MICROVESSELS DENSITY IN GASTRIC MUCOSA OF DYSPEPTIC PATIENTS A. G. McNicholl, *,  M. E. Fernandez-Contreras, *,  P. M. Linares, *,  A. C. Marin, *,  C. Lopez-Elzaurdia,* M. Chaparro *,  and J. P. Gisbert *,  *La Princesa University Hospital, Madrid, Spain;   CIBERehd, Spain Introduction: Published data on the role of H. pylori in angiogenesis are con- tradictory, while its role in lymphangiogenesis has not been investigated. Aims: To investigate the density of lymphatic and blood vessels, together with H. pylori infection, in biopsies of gastric body and antrum of dyspeptic patients. Methods: Biopsies from patients subjected to gastroscopy according to clinical criteria were studied by immunohistochemistry. Exclusion criteria were: previous eradication therapy, ulcer disease, regular non-steroidal anti-inflammatory use, and proton pump inhibitor or antibiotic treatment 15 and 30 days prior to recruitment respectively. Microvessel density was determined with CD34 and D2.40 monoclonal antibodies (DAKO, Glostrup, Denmark), which are markers of blood and lymphatic vasculature respectively. Quantification was performed by direct counting of microvessels in four fields at 40· magnification. H. pylori infection was assessed by 13 C-urea breath test and/or histopathological diagnosis. Results: Twenty-three biopsies (13 antral) from 13 patients were studied. Their median age was 62 years, 46% males, 73% gastritis and/or atrophy and 24% H. pylori positive. Higher blood microvessels densitiy was associated to H. pylori infection (p = .03) (Table 1). Microvessels count was slightly elevated, without statistical signification, among biopsies with histological diagnosis of gastritis. Further associations with inflammatory activity or location were not found. Lymphangiogenesis was not related with any of the studied variables. Conclusions: (1) H. pylori infection was associated with increased gastric angiogenesis (2) Lymphangiogenesis was not related with the studied clinico- pathological features. Abstract no.: P3.7 COX-2 INHIBITION WITH NUTRACEUTICALS: A NEW THERAPEUTIC APPROACH AGAINST HELICOBACTER PYLORI INFECTION? A. M. Santos,* M. Oleastro,   T. Lopes,* T. Pereira, à E. Seixas, § P. Chaves, à J. Machado § and A. S. Guerreiro* *CEDOC/Faculdade de Cie ˆ ncias Me ´ dicas, Univ. Nova de Lisboa, Lisboa, Portugal;   Instituto nacional de Sau ´ de Dr. Ricardo Jorge, Lisboa, Portugal; à Instituto Portugue ˆs de Oncologia – Lisboa, Lisboa, Portugal; § Instituto Gulbenkian de Cie ˆ ncia, Oeiras, Portugal Accumulated evidence in humans and animals shows that H. pylori up-regulate the expression of cyclooxygenase (COX)-2 both at mRNA and protein levels which might be the one of the mechanisms leading to several gastric diseases. Aim: To study the expression of COX-2 on mice gastric mucosa during long-term treatment with two nutraceuticals: curcumin and synbiotic 2000 Ò on H. pylori experimental chronic infection. Materials and Methods: We infected 45 C57BL/6 mice with SS1 – H. pylori strain. After infection confirmation by 13 C-urea breath test mice where then treated with either PBS, curcumin (10 mg/mouse) or Synbiotic 2000 Ò (50 mg/ mouse), three times per week. Five mice from each treatment group were euthanized at week 6, 18 and 27. Gastric samples were removed for COX-2 immunohistochemistry analysis. Results: All the 45 mice were Hp positive by 13 C-urea breath test and immu- nohistochemistry. In the PBS group the production of COX-2 was significantly up-regulated at week 6 (area of positive immunostaining 393–544 · 10 3 pixels), 18 (area of positive immunostaining 242–614 · 10 3 pixels) and 27 week (area of positive immunostaining 129–175 · 10 3 pixels). The treatment with either curcumin or synbiotic significantly decreased the expression of COX-2 at all time points. Conclusions: These results suggest the therapeutic usefulness of both nutra- ceuticals on COX-2 inhibition during chronic experimental mice H. pylori infec- tion. The supplementation of diet in humans with curcumin or Synbiotic 2000 Ò may be a novel therapeutic approach against gastric inflammation induced by Hp infection. Abstract no.: P3.8 PRODUCTION OF ANTI-H. PYLORI IMMUNOGLOBULIN Y (IGY) IN THE CHICKEN EGG YOLK P. Saniee,* F. Siavoshi,* G. Nikbakht Broujeni,   M. khormali   and A. Sarafnejad à *Department of Microbiology, Faculty of Sciences, University of Tehran, Tehran, Iran;   Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran; à School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Introduction: Immunizing chickens with certain antigens and collecting the related antibodies have been proposed as a useful method for production of edible immunoglobulins (Igs). Oral IgY could have applications in the control of gastric infections such as those caused by Escherichia coli and Salmonella enterica. In this study immunization of chicken and production of anti-H. pylori IgY in egg yolk was studied Methods: A suspension of one heat-killed and PCR-confirmed H. pylori isolate mixed with freund’s adjuvant was intramuscularly injected to two 5 months-old chickens, once a week for three consecutive weeks. Ten days after the last immunization, eggs were collected and total antibody was purified from egg yolk. The presence of anti-H. pylori IgY was assessed using dot blotting method on Polyvinylidene fluoride membrane. PCR-confirmed E. coli and Salmonella entrica were used to eliminate the possibility of cross-reaction. Results: Specific binding of extracted antibodies to H. pylori-specific antigens was observed as colored spots on the membrane, indicating the presence of anti-H. pylori ploycolonal antibodies in the chicken eggs. Colorimetric reaction of anti- bodies with E. coli and S. entrica-specific antigens were not observed. Discussion: Immunization of chickens for the production of antibodies has several advantages such as no need for blood sampling from animals and pro- duction of a large amount of antibodies in egg yolk. Furthermore, egg yolk containing anti-H. pylori antibody administered orally could provide a novel and effective approach to prevent H. pylori infection. 98 ª 2012 Blackwell Publishing Ltd, Helicobacter 17 (Suppl. 1): 65–120 Abstracts