167 Advanced Glycation End Products Are Immunodominant Targets of Multi-Specific Natural Antibodies Miho Chikazawa1, Hiroaki Miyashita1, Takahiro Shibata1, Yoshichika Kawai1, and Koji Uchida1 1Nagoya University, Japan Advanced glycation end products (AGEs) are a heterogeneous, complex group of compounds that are formed when reducing sugars, such as dehydroascorbic acid (DHA), react with amino acids in proteins and other macromolecules. The presence and accumulation of AGEs in many different cell types affect cellular structure and function. We now show that the AGEs are an immunodominant target of natural antibodies that recognize multiple antigens, including double-stranded DNA and apoptotic cells. Prominent IgM titers to the DHA-derived AGEs were detected in the sera of normal mice and were markedly accelerated by the immunization with the AGEs. In addition, a progressive increase in the IgM response against the AGEs was also observed in SLE-prone mice. Strikingly, a subset of the anti-AGEs IgM monoclonal antibodies were specifically bound to apoptotic cells in vitro and tissue cells in vivo. These findings suggest that natural antibodies may ubiquitously recognize AGEs as one of their targets and contribute to the protection against multiple damage-associated molecules.

168 Glutathione Peroxidase-1 and -2 Protect Mice from Salmonella-Induced Colitis Robert Steven Esworthy1, Byung-Wook Kim1, James H Doroshow2, Thomas L Leto3, and Fong-Fong Chu4 1Beckman Res Inst of City of Hope, 2National Cancer Institute, 3National Institute of Allergy and Infectious Diseases, 4Beckman Research Institute of City of Hope Salmonella enterica Serotype Typhimurium (S. Ty) is a major source of food borne gastroenteritis or food-poisoning. Much is known about S. Ty, which are Gram-negative, facultative intracellular bacteria. A large region of the S. Ty genome encodes proteins that evade host defense systems, cause invasion of host cells and promote intracellular bacterial trafficking and replication. Little is known about host defense mechanisms. Recent studies have shown that the inflammatory bowel disease (IBD) susceptibility genes, NOD1 and NOD2, play important anti-bacterial roles in eukaryotes. NOD1 and NOD2 are two intracellular bacteria-sensor proteins, which activate autophagy to kill the invading S. Ty. Since autophagy is induced by reactive oxygen species (ROS) and antioxidant enzymes and antioxidants can decrease ROS levels and autophagy, we tested whether mice deficient in GPX1 or GPX2, two intracellular GPXs, are more resistant to S. Ty infection. We inoculated 2x107 S. Ty to wild-type (WT) C57BL/6J (B6) and 129S1Svlm/J (129) mice as well as 129 Gpx1-KO and Gpx2-KO mice, which had been pretreated with metronidazole for four days to allow bacterial colonization. Instead, we found both Gpx1-KO and Gpx2-KO mice are highly susceptible to S. Ty colonization and colitis, while WT B6 and 129 mice are resistant to S. Ty infection analyzed four days after oral inoculation of S. Ty. We also found a Gdac1 (Gpx-deficiency-associated colitis 1) locus, which modulates the severity of spontaneous colitis in Gpx1 and Gpx2-double knockout (Gpx1/2-DKO) mice, impacts S. Ty-induced colitis in the Gpx1-KO and Gpx2-KO mice. Since our results suggest that both GPX-1 and

GPX-2 protect mice from S. Ty-induced colitis, whether GPX deficiency affects S. Ty infection or autophagy will be investigated further. The Gdac1 locus was identified from studying B6 and 129 Gpx1/2-DKO mice, which have mild and severe colitis respectively. The impact of Gdac1 locus on S. Ty-induced colitis in non-DKO mice substantiates its role in innate immunity.

169 Inflammation and Histamine Intolerance in Children with Crohn’s Disease: Influence of Polyphenolic Extract, Pycnogenol Zdenka Durackova1, Marian Kolacek1, Monika Dvorakova1, Zuzana Paduchova1, Jana Muchova1, Iveta Cierna2, Dagmar Szekyova1, Ingrid Zitnanova1, and Laszlo Kovacs1 1Comenius University, Slovakia 2Childrens University Hospital, Slovakia Crohn’s disease (CD) is an autoimmune inflammatory bowel disease. The pathophysiology of CD is still quite unknown. Pycnogenol (Pyc), standardized extract from the bark of Pinus pinaster, is a mixture of polyphenolic compounds composed of catechin, epicatechin, taxifolin and procyanidins. Pyc acts as an antioxidant in vitro and biomodulator of redox state in many diseases. The aim of our study was to examine influence of Pyc on oxidative stress, inflammation and histamine intolerance in CD children. Fifteen children (aver. age 16.3 years) with CD (7 males (M), 8 females (F)) and 15 healthy children (aver. age 14.3 years, 6 M and 9 F) were included in the study. CD children were administered with Pyc (2 mg/kg) for 10 weeks. Children were examined before Pyc administration, 5 and 10 weeks after the start and 2 weeks after intervention. The clinical state was monitored by PCDAI (Pediatric Crohn´s Disease Activity Index), inflammatory activity by CRP and calprotectin (CP) levels. Also some parameters of oxidative stress (OS) and the activity of diamine oxidase (DAO) characterizing histamine intolerance were determined. Patients with CD were in remission state, their PCDAI index during 10 weeks of Pyc administration was not significantly changed (3.93 – 4.85). CD patients in the contrary to controls had increased level of CRP (12.63 vs. 1.2 mg/L) and calprotectin (976.16 vs. 21.28 μg/g stool). Pyc administration did not influence inflammatory parameters. The activity of DAO in CD patients at the beginning was not different from controls. Pyc administration after 5 weeks increased DAO activity. Acknowledgment: This work was partly supported by grant of Ministry of Health of SR 2007/16-UK-01 and Mind & Health, civil association. Pycnogenol was kindly provided by Horphag Res. Ltd, Geneva, Switzerland.

170 Defining the Effect of Novel Slow-Release H2S Donors on Pro-Inflammatory Mediators and Enzymes in Human Joint Cells B Fox1, T Holland1, A Perry2, PG Winyard1, ME Wood2, and M Whiteman1 1University of Exeter Medical School, UK 2University of Exeter, UK Elevated levels of the gaseous mediator hydrogen sulfide (H2S)

SFRBM 2012 S75

doi:10.1016/j.freeradbiomed.2012.10.223

doi:10.1016/j.freeradbiomed.2012.10.224

doi:10.1016/j.freeradbiomed.2012.10.225