11:10-12:00, Rm 103

ImmunologyO-18-14IL-18 is an important target for development of immune modulator inpulmonary diseasesDo-Hyun Kim¹ , ², Min-Jong Kang³, Je-min Choi¹ , ² , *¹Department of Life Science, Hanyang University, ²Hanyang Biomedical Research Institute, Seoul133-791, Korea and ³Section of Pulmonary and Critical Care Medicine, Yale University School ofMedicine, CT 06520, USAInterleukin-18 (IL-18), originally identified as IFN-γinducing factor and it was known toplay important roles in various autoimmune and allergic diseases such as COPD, RA,and asthma, etc. However the importance of IL-18 as a target of immune modulator, isnot much appreciated. Here we investigated that IL-18 induces Th1, 2 and 17inflammation in the lung including alveolar destruction and airway inflammation in lungspecific IL-18 TG mice. To develop IL-18 antagonizing immune modulator, we havegenerated novel CPP conjugated IL-18BP chimeric protein (CPP-mIL-18BPc). IL-18BP isa naturally occurring IL-18 antagonizing protein and CPP-mIL-18BPc efficiently deliveredinto cells time and dose dependent manner. Treatment of CPP-mIL-18BPc inhibitedmacrophage activation potentially suggesting that IL-18 targeting immune modulatorwould be a attractive novel therapeutic approach to regulate immune responses. Theseresults demonstrate that IL-18 plays an important roles in lung destruction and pulmonaryinflammation and CPP-mIL-18BPc can be a novel immune modulatory protein in variousinflammatory diseases. [Supported by grants from KRF 2010-0025389]O-18-17Anti-inflammatory effects of SHRC in DSS-induced colitis modelHye - Eun Choi¹ , ², Seung jun Lee¹ , ², Hyo Jin An¹and Kyung-Tae Lee¹ , ² , *¹Department of Pharmaceutical Biochemistry, College of Pharmacy, Kyung Hee University 1 Hoegidong,Dongdaemun-gu, Seoul 130-701, Korea, ²Department of Life and NanopharmaceuticalScience, College of Pharmacy, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, KoreaIn this study, we studied that alleviatory effect of SHRC (saponin- high fraction of Rubuscoreanus Miquel) in DSS-induced colitis model and the molecular mechanisms of SHRCfor its anti-inflammatory properties in LPS-induced RAW 264.7 macrophage cells. In DSScolitis model, the DAI score and detection of MPO accumulation revealed SHRCsignificantly inhibited loss of body weight, suppression of diarrhea and bleeding, andinfiltration of macrophage. Moreover, H&E staining also indicated SHRC suppressed thethickness of muscle layer, edema, mucosal damage by DSS. In addition, SHRCdecreased the nitric oxide (NO) production in DSS-induced colitis model. SHRCattenuated the DSS-induced pro-inflammatory cytokines. SHRC suppressed the LPSinducedNO and prostaglandin E2 (PGE2) production and pro- inflammatory cytokines.We determined that SHRC inhibited the transcriptional activity of nuclear factor-kappa B(NF-κB) and its translocation to the nucleus though the suppression of inhibitory kappa Bα(IκBα) phosphorylation in LPS-activated RAW 264.7 cells. Taken together, these resultssuggest that the anti-inflammatory effect of SHRC is achieved through the inhibition ofiNOS, COX-2, pro-inflammatory cytokines expression via suppression of NF-κB activationin vivo and in vitro.O-18-15Farnesoic O-methytransferase-like protein of Tenebrio molitor larvae isspecifically recruited on Mycobacterium marinum cellsYong-In So¹, Lili Zheng¹, Ye-Rang Heo¹, Seong-Min Kang¹, Yeo-Jin Won¹, Dong-Jun Jung¹, Yong-Hun Jo¹, Kenji Kurokawa¹, Ki-Hye Kim², Eun-kyeong Jo²andBok-Leul Lee¹¹The national Research Laboratory of Defense Proteins, College of Pharmacy, Pusan NationalUniversity, Kumjeong Ku, Busan 609-735, Korea, ²Department of Microbiology, College ofMedicine, Chungnam National University, 6 Munhwa-dong, Jungku, Daejeon 301-747, KoreaMycobacterium tuberculosis is a major intracellular pathogen of humans, with anestimated one-third of the global population being latently infected, 2-3million deathsannually from tuberculosis, and an increasing numbers of multidrug-resistant strains totuberculosis. Mycobacterium marinum, a close relative of Mycobacterium tuberculosis isbeing used to study mycobacterial pathogenesis. Here, we found that antimicrobialpeptides (AMPs) were induced in Tenebrio molitor by injection of Mycobacteriummarinum into Tenebrio larvae. We also observed that a 19 kDa protein originated fromTenebrio hemocytes specifically recruited on the Mycobacterium marinum cell surface.When we obtained cDNA of 19 kDa protein, it showed high homology with that offarnesoid O-methyltransferase-like protein (OMT) of Tribolium cataneum. When dsRNAof Tenebrio OMT was injected into T. molitor larvae, OMT gene expression was knockdown.Now we are determining the biological function of OMT protein duringMycobacterium marinum infection.O-18-18Anti-wall teichoic acid antibodies in human serum specifically activatehuman classical complement pathwayDong-Jun Jung, Jang-Hyun An, In-Kyung Seo, Min-Su Kim, Kyoung-Hwa Ryu,Yong-In So, Yong-Hun Jo, Kenji Kurokawa and Bok Luel LeeFrom the National Research Laboratory of Defense Proteins, College of Pharmacy, Pusan NationalUniversity, Busan 609-735, KoreaStaphylococcus aureus are covered with several cell wall components, such aspeptidoglycan (PGN), lipoteichoic acid (LTA), and wall teichoic acid (WTA). Thesemolecules are known to function as ligand molecules during activation of innate immuneresponses. But, their exact biological roles are not clearly determined. We recentlydemonstrated that WTA of S. aureus is a functional ligand of mannose-binding lectin(MBL), a serum mediator of lectin complement pathway However, the molecular functionof S. aureus WTA during human complement activation is not clearly demonstrated. Toexamine the function of WTA, we purified anti-WTA antibodies and anti-LTA antibodiesfrom human intravenous immunoglobulin (IVIG) and then checked the abilities bymeasuring C4 and C3 deposition. Anti-WTA antibodies specifically deposited C4 on S.aureus cell surface, but not anti-LTA antibodies, suggesting that anti-WTA activateclassical complement pathway. To further confirm whether anti-LTA antibodies canactivate alternative complement pathway, we examined the C3 deposition using C2deficient and factor B-deficient serums. These results demonstrated that the cell wallcomponents of S. aureus function as critical ligand molecules during human complementactivation.O-18-16Comparison of immunological properties about monomeric dap-typepeptidoglycan between tenebrio molitor and trbolium castaneum by rnainterference experimentJun-Ho Chae, Bing Zhang, Eun-Han Park, Jang-Hyun An, Dong-Jun Jung, Yong-InSo, Yong Hun Jo, Kenji Kurokawa and Bok Luel LeeFrom the National Research Laboratory of Defense Proteins, College of Pharmacy, Pusan NationalUniversity, Busan 609-735, KoreaBacteria peptidoglycan(PGN), a bacterial cell wall component, is known to be recognizedby peptidoglycan recognition protein(PGRP), which are acting as pattern recognitionmolecules during insect innate immunity. Different types of PGNs are known to berecognized by different PGRPs during insect innate immune responses and induce theproduction of antimicrobial peptides. Previously, our laboratory reported the recognitionmechanism and its signaling pathway of polymeric DAP-type PGN in Tenebrio molitorlarvae. However, the molecular recognition mechanism of monomeric DAP-type PGN isstill unclear. Recently, our group found a novel 17 kDa protein from Tenebrio hemolymphthat showed high sequence homology with that of Drosophila PGRP-LE. To furtherunderstand the molecular recognition mechanism of monomeric DAP-type PGN, we havetried to RNA interference experiment of Tenebrio newPGRP homolog gene(Tc-PGRP1,2)using the red flour beetle, Tribolium castaneum. we checked significant knockdown oftarget genes by injecting dsRNA and then proceeded to inject different types of PGNs. Asa result, we observed that induction of several kind of AMPs significantly decreased onlyin monomeric-DAP PGN injected Tribolium larvae but not in Lys-type PGN injected larvaeafter knockdown of PGRP.O-18-19Immune response in tenebrio molitor larvae by invasion ofacholeplasma laidlawiiSeong-Min Kang, Yeo-Jin Won, Ho-Jung Eo, Yong-In So, Dong-Jun Jung, YongHun Jo, kenji kurokawa, Bok-Luel LeeFrom the National Research Laboratory of Defense Proteins, College of Pharmacy, Pusan NationalUniversity, Busan 609-735, KoreaPeptidoglycan, a major cell wall component of bacteria, is known to trigger strong innateimmune responses in the insects and human. However, Mycoplasma species belongingto a class of Mollicutes is lack of peptidoglycan in their cell wall. However, Mycoplasmaspecies, such as Mycoplasma pneumoniae and Mycolpasma genitalium, are majorpathogen causing pneumonia in human and animals. Here, we tried to purify whichmolecule(s) of Mycoplasma species is involved in Mycoplasma-mediated pathogenesis.When we injected Acholeplasma laidlawii, which is culturable strain in the laboaratory,into coleopteran insect Tenebrio larvae larvae, antibacterial activity was induced againstEscherichia coli and Staphylococus aureus, but peptidoglyca-mediated melanin was notsynthesized. To purify the molecule showing antibacterial activities to homogeneity, weperformed several column chromatographies. Finally we obtained three moleculeseliciting antibacterial activities by C18 reverse phase HPLC. Now we are determining itsstrctures using NMR and Mass analysis.306 Korean Society for Biochemistry and Molecular Biology