TY - JOUR
AB - PP-086-09 Enhanced phosphorylation of STAT-3 Serine 727 and propagation of dynamic alkaline phosphatase during calcification in culture of HUVEC J. Mendoza J. Mendoza National University of Mexico, Mexico City, Mexico J. Caceres J. Caceres National University of Mexico, Mexico City, Mexico Abstract Vessel calcification involves an active cellular mineralization process. The calcification of smooth muscle cells depends on molecules supplied by the endothelium. However, the pathways and cellular events by which calcification occur in vascular endothelial cells have not been sufficiently studied. We hypothesized that calcifying conditions in vitro would compose specific signals in endothelial cells. Methods and Results: We performed experiments that evaluated the effect of osteogenic medium (OM) on alkaline phosphatase activity, signaling pathways and cell death. We found that OM emits a survival signal at the beginning of cell culture and then causes massive cell death via autophagy. As calcification progresses initial intracellular vesicular calcification leads to the complete calcification of large cellular compartments, whereas unstimulated HUVEC do not calcify. The extent of calcium deposition was measured using Alizarin Red S staining. We observed that it is a process potently controlled by propelled invasive alkaline phosphatase (ALP) produced in stimulated cells. To better understand the signaling networks involved, we analyzed the HUVEC- and HUVEC-treated cell lysates using the Kinexus™ KPSS 1.3 phospho-site broad coverage pathway screen. Our studies revealed that STAT3, JNK, B23 (NPM), and Rb mediate survival and calcification in HUVEC. Conclusion: These data suggest that under calcifying conditions, vascular calcification is mainly a STAT3/ALP/autophagy-mediated process. This work was supported by grant PAPIIT IN221309 PP-086-10 Catalitic antibodies in autoimmunity. Development of catalytic vaccines A. G. Gabibov A. G. Gabibov Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation Abstract Discovery of catalytic antibodies (abzymes) was a revolutionary event that created new junctions between chemistry, biochemistry, immunology and pathology. Creation of abzymes is based on the intrinsic properties of immunoglobulin superfamily to produce complementary “molecular imprints”. These “catalytic imprints” could be made from the stable chemical analogs of transition state (TSA) of the enzyme reaction. This approach was successfully developed by Richard Lerner group. The alternative way to create abzymes was proposed by author. This allowed us to generate abzymes with acetylcholinesterase and protease activities. We used also autoimmune mice models to create epitope-specific abzymes.This may give rise to biocatalysts with new functions, previously unknown for common enzymes, which may be very profitable for fine organic synthesis. This method stimulated our attempts to make antibody-like acceptors for phosphorus-based poisons. Recombinant antibodies with such functions were obtained in this lab using chemical selection of “naïve” phage-display library. The X-ray of this antibody was made and 3D structure was solved. Other advantage of abzyme field is the opportunity to make “catalytic vaccines”. Numerous attempts to combat HIV infection using drug therapy as well as classical vaccination turned out to be ineffective. One of the targets for the novel therapeutic approach may be the main surface antigen, viral envelope protein gp120. The specific cleavage of this protein can lead to the dramatic changes in the immune response toward virus and decrease binding of HIV to CD4 receptor. PP-086-11 Silver nanoparticles induce developmental toxicity in zebrafish (Danio rerio) via ROS generation D. Lim D. Lim 1 Department of Microbiology, National Core Research Center for Nano Medical, Yonsei University, Seoul, Republic of Korea T. Kang T. Kang 2 Department of Chemical Engineering, Yonsei University, seoul, Republic of Korea K. Lee K. Lee 2 Department of Chemical Engineering, Yonsei University, seoul, Republic of Korea S. Park S. Park 3 Department of Internal Medicine, Yonsei University, seoul, Republic of Korea I. Choi I. Choi 1 Department of Microbiology, National Core Research Center for Nano Medical, Yonsei University, Seoul, Republic of Korea Abstract Silver nanoparticles (AgNPs) induced cytotoxicity depends on particle size. To evaluate nanotoxicity we have used zebrafish model, in which genotoxicity and development toxicity can be assessed in vivo. To investigate whether the effect of AgNPs induce developmental toxicity, fertilized eggs from zebrafishes were treated with AgNPs of 5 nm. Our results suggest that the mortality rate of embryos is dependent on the concentration of AgNPs. At 10 ug/mL of AgNPs, all the embryos could not survive. In accordance with mortality rate the hatch rate decreased. Even among live and hatched fishes 5 nm AgNPs induced developmental anomalies such as bent spines, pericardial edema, tail anomaly and persistent yolk sac. Using acridine orange, we found apoptosis was induced in fishes with treatment of AgNPs. More interestingly, zebrafish exposed AgNPs showed neurotoxicity and damage of developing nerve cells. Our results performed with staining by CM-H2DCFDA or ROS scavenger (NAC:N-acetylcysteine) clearly indicate that ROS is responsible for these developmental defects. This work was supported by NRF through National Core Research Center for Nanomedical Technology (R15-2004-024-00000-0) PP-086-12 Identification of differentially expressed proteins in B-MD-C1 and B-MD-C1 (ADR+/+) by proteomics C. Zhang C. Zhang Hebeisheng, Shijiazhuang, China W. Lihua W. Lihua Hebeisheng, Shijiazhuang, China A. Jun A. Jun Hebeisheng, Shijiazhuang, China S. Baoen S. Baoen Hebeisheng, Shijiazhuang, China Abstract Chemotherapy plays an important role in the comprehensive therapy of cancer. However, the drug-resistance often causes the failure of the chemotherapy. The aim of this study is to investigate differently expressed protein B-MD-C1 and B-MD-C1 (ADR+/+) cell by proteomic analysis. Methods: In the first part of our experiment,protein separation was performed using two-dimensional gel electrophoresis(2-DE) with Amersham 18 cm linear pH 3-10 immobilized pH gradient(IPG) strips. The images of the gels were analyzed by PD-quest 7.0 software to find the differentially expressed protein-spots in each group. Then the differentially expressed protein-spots were incised from the gels and digested by trypsin. The peptide mass fingerprinting (PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The proteins were identified by data searched in the Mascot-database. Results: Two-dimensional protein maps of B-MD-C1 and B-MD-C1 (ADR+/+) were gained successfully. Two hundred and seventeen differentially expressed proteins were screened by analysis the electrophoretic maps of B-MD-C1 and B-MD-C1 (ADR+/+). Thirty-six differential proteins were analyzed by peptide mass fingerprinting. Heat shock protein beta-1, tubulin beta-2C chain, eukaryotic translation initiation factor, heterogeneous nuclear ribnucleoprotein h and ribosomal protein were identified. All four differentially expressed proteins were over-expressed in B-MD-C1 (ADR+/+), whereas low-expressed or no-expressed in B-MD-C1. Conclusion: The 2-DE gels images are successfully acquired from B-MD-C1 and B-MD-C1 (ADR+/+) cell. These differentially expressed proteins give some clues to elucidate the mechanism of tumor cell resistant of cisplatin, providing the basis of searching for potential target of chemotherapy of cancer. PP-086-13 Suppressive effects of intravenous immunoglobulin (IVIG) on human umbilical cord blood immune cells J. Qian J. Qian Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China T. Chen T. Chen Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China M. Wang M. Wang Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China S. Wu S. Wu Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China Abstract We investigated in vitro immunomodulatory effects of intravenous immunoglobulin (IVIG) on cord blood mononuclear cells (CBMNC), macrophages, dendritic cells and CD3+ T cells isolated from umbilical venous blood. Cell proliferation used 3H-TdR incorporation, culture supernatants were assayed for cytokines using ELISA and surface marker expressions were determined by flow cytometry. IVIG suppressed CBMNCs and CD3+T-cells proliferation, secretions of IL-10, INF-γ and TGF-β1, but not IL-4, and PHA-induced expressions of surface molecules (CD25, CD45RA and CD45RO), with more pronounced effects for CBMNCs. IVIG decreased cord blood (CB) macrophage phagocytosis and CD14, HLA-DR and CD86 expressions. IVIG increased CD14 expression and decreased MCH II expression for differentiation-stage CB dendritic cells (DCs) and increased CD14 expression and decreased CD80 and CD83 expressions of mature DCs, suggesting that IVIG intervention inhibited DC differentiation and maturation. In addition to T cells, IVIG immunomodulatory effects on CBMNCs involve a variety of cells and molecules. CB macrophages and CBMNC-DCs are targets of IVIG. PP-086-14 Immunological features of fullerene C-60 and its derivatives Y. N. Bashkatova Y. N. Bashkatova National Research Centre Institute of Immunology FMBA of Russia, Moscow, Russian Federation S. M. Andreev S. M. Andreev National Research Centre Institute of Immunology FMBA of Russia, Moscow, Russian Federation A. O. Petrukhina A. O. Petrukhina National Research Centre Institute of Immunology FMBA of Russia, Moscow, Russian Federation A. A. Babakhin A. A. Babakhin National Research Centre Institute of Immunology FMBA of Russia, Moscow, Russian Federation Abstract Background: The increased use of fullerene materials in cosmetic, biomedical applications is an occasion for a detailed examination of their health effects including the immunological ones. In the present work we tried to generate fullerene-specific antibodies to design the C60-specific ELISA. Other purpose was to determine the adjuvant effect of fullerene derivatives (FDs). Methods: Pristine fullerene in form of aqueous nanodispersion and FDs with functional addends: ε-aminocaproic acid (C60-Acp), maleimide-proline, malonic acid and tris-malonic acid, conjugates of C60-Acp with BSA, KLH and ovalbumin were used as immunogens in mice and antigens in ELISA including competitive ELISA. Results: The data obtained suggest that the immune system can respond to an immunization with the fullerene-protein conjugates by the generation of IgM and IgG class antibodies toward protein epitopes and complex epitopes consisting of FDs and protein parts. Modification by FDs most probably results in conformational changes in protein creating the cryptic epitopes. A competitive ELISA showed that anti-C60-protein antibodies do not interact to nano-C60. The nano-C60, used as immunogen together with Friend's adjuvant, was not capable to induce C60-specific IgG and IgE antibodies. The FDs used as stimulating additive to protein immunogens did not stimulate anti-protein IgG responses (BSA, KLH, OVA). Conclusion: In all likelihood, the purely carbonic fullerene is unable to mount specific epitope. Fds examined here do not stimulate or suppress the immune responses to proteins. Taking in consideration of good membrane penetration ability and the absence of immunogenicity nanoparticle FDs are potentially useful measure for drug and vaccine delivery. PP-086-15 General and local levels of IL-2 and IL-6 in patients with primary open angle glaucoma and in patients of glaucoma risk grope D. Rukina D. Rukina Vladivostok State Medical University, Vladivostok, Russian Federation E. Markelova E. Markelova Vladivostok State Medical University, Vladivostok, Russian Federation Abstract Glaucoma is one of the leading reasons of visual loss and blindness in the world. Pathogenesis of glaucoma is still discussing and is not understood totally. The recent researches demonstrate the meaning of immune mechanisms in glaucomatous optic neuropathy (GON). Cytokines are known to be the main mediators of cells relations and realize all the immune processes. IL-2 plays a great role in development of cell and humeral immune reactivity. Its main function is to guarantee adequate immune answer. It also saves cells of apoptosis. IL-6 refers to neuropoetines and supports stability of neurons. We have studied the levels of 2 and 6 (IL-2, IL-6) in serum and in tear of 23 glaucoma patients. The age of the patients waved from 40 to 75 years. The levels of cytokines were detected by IFA-method. The IL-2 level was 3 times increased in all the patients in serum (8,68±3,2pg/ml, the normal level 2,6±0,3 pg/ml,<0,05), in tear it showed 38 times increasing (3,82±1,6 pg/ml, the normal level 0,12±0,1 pg/ml,<0,01). IL-6 was increased in two times in serum (1,93 ±0,2 pg/ml, the normal level 0.87±0,4 pg/ml,<0,01). It turned out, that investigation of IL-2 local level was more informative in diagnostic of immune misbalance, than IL-2 systemic level. Therefore, glaucoma patients reveal dysfunctional changes in immune system, which is an important link in the mechanisms of the systemic immune response during GON and needs immunocorrection. PP-086-16 The comparison of immune response factors in exercisers and non-excercisers A. Khosravi A. Khosravi Ilam university of Medical Sciences, Immunology Dep, Ilam, Islamic Republic of Iran H. Peyman H. Peyman Ilam university of Medical Sciences, Immunology Dep, Ilam, Islamic Republic of Iran F. Soleimani F. Soleimani Ilam university of Medical Sciences, Immunology Dep, Ilam, Islamic Republic of Iran M. M. Nahal M. M. Nahal Ilam university of Medical Sciences, Immunology Dep, Ilam, Islamic Republic of Iran Abstract Introduction:Exercise in different situation and with different quality can change the Immune response accordingly. Many studies documented that systemic exercise can support the immune responses while discontinues exercise weakens it. The current study was designed to analyse the Immune properties amongst exercisers compared to non exercises. Material and Methods: 20 exercisers as case group together with 20 non exercisers as control group were randomly selected from each 6ml blood was taken after 90 minutes exercise was ordered. CD3, CD4, CD8, CD19, CD56 and IgG titter were all measured using flowcytometry and Nephlometry analyses. Results: The mean CD3 level was 62.64 and 69.5 in men and women of case group and totally 64.33 in control group. The mean CD4 level was 41 for men, 41.83 for women and 42.86 for control group though the mean CD8 level for men was 19.5, women 27 and control group 20.33. The level of CD19 was 11 for men, 9.6 for women and finally 13.6 for control group. The mean value of CD56 was 7.92 for men, 7.83 for women and 6.39 for control group. The mean titer of IgG was 14.09, 14.49 and 14.14 for men, women and control group respectively. Conclusion: As the mean OD value of CD3 and CD8 and the CD4/CD8 ratio are all significantly different among exercisers than non-exercisers indicating that immune system can be affected dependent on the nature and quality of exercise. PP-086-17 Inhibition of Carrageenan-induced edema by Tripleurospermum Disciforme extracts in rats A. Bakhtiarian A. Bakhtiarian School of Medicine, Tehran University of Medical Sciences, Tehran, Iran, Tehran, Islamic Republic of Iran Abstract Anti-inflammatory effect of Tripleurospermum disciforme extract was studied in rats. The effect of the extract against acute inflammation was studied by hind paw edema test. Intraperitoneal injection of different doses (15mg/kg, 30mg/kg, 45mg/kg, 60mg/kg, 90mg/kg, 120mg/kg) of Tripleurospermum disciforme which was followed by hind paw carageenan injection 1 hour later were investigated. The resultant edema was quantified by measuring changes of diameter of hind foot. After 2, 3, 4 and 6 hours we measured the anti-inflammatory effect of Tripleurospermum disciforme extract. Tripleurospermum disciforme extract with doses of 45mg/kg, 60mg/kg, 90mg/kg and 120mg/kg and Indomethacin 5mg/kg showed significant effect. It is concluded that Tripleurospermum disciforme extract has anti-inflammatory effect against acute inflammation. PP-086-18 Flocytometric evaluation of B-lymphocytes (CD19+) in peripheral blood of healty smokers and non-smokers K. Z. Entezami K. Z. Entezami Iran University of Medical Sciences, Tehran, Islamic Republic of Iran Abstract Objcetive : The purpose of this work was to evaluate the normal B-lymphocyte phenotype in the peripheral blood, was carried out in 40 untreated healthy nonsmoking volunteers and in 35 cigarette smoking, aged between 18-60 years. Methods : Anticoagulated peripheral blood was stained with monoclonal antibodies and B-lymphocytes were analyzed by flowcytometry for the expression of the surface antigen of peripheral blood B-lymphocytes (CD19+) in smokers. Results : There was a decrease in the total count of CD19+ B- lymphocytes in smokers (12.29± 3.2) compare with non-smokers (13.89±4.72), Finally, the mean percentages of B cells in smoker subjects revealed a significant decrease with advancing age (P<0.05). Data were analyzed by using spss software. Conclusions : We suggest that these findings may provide further insight into the association of smoking with an increased incidence of certain malignant diseases and respiratory infections . Therefore these observations have important clinical implications for the use of B-lymphocytes measurement in Iran, especially in the management of viruse infections . PP-086-19 Evaluation of continuously intensive exercise effect on T cell and expression of HSP72 Z. Shaker Z. Shaker Immunology Dep. Faculty of Medicine, AJA University of medical Science, Tehran, Islamic Republic of Iran A. Goodarzi A. Goodarzi Immunology Dep. Faculty of Medicine, AJA University of medical Science, Tehran, Islamic Republic of Iran M. Shohoodi M. Shohoodi Immunology Dep. Faculty of Medicine, AJA University of medical Science, Tehran, Islamic Republic of Iran Abstract The continuously intensive exercise can effect immune system as showed on last studies. Concerning to role of adaptive cell mediated immunity against virus and deletion tumor cell, we decided to evaluate T cell and HSP72 (Heat Shock Protein72). In an analytical research, we took 5cc of peripheral blood of 150 people before and after intensive exercise for assessing of complete blood count and T cell number and function by CD3/CD4, CD8 and CD25, respectively. Also human HSP72 of serum samples were evaluated by ELISA in 10 persons. We Analyzed findings by paired t test and non-parametric willcoxon in SPSS 16 ver. Software. The flow cytometric analysis showed that the number of CD3+/CD4+ T+ cell was increased the number of CD8+ was significantly decreased after exercise compared to before (Pv<0.05). T cell was activated significantly after continuously aerobic and non- aerobic exercise (Pv<0.05). hHSP72 was not significantly increased after exercise (Pv<0.05). In our study, we should mention that the number of CD3/CD4 and CD8 T cells were in normal range. It seems the number of samples was not enough to show a significant result of hHSP72. For evaluating briefly, we need to increase the number of samples. PP-086-20 Analysis on SHP2-interacting transmembrane adaptor protein (SIT) in lymphocytes development Y. Tsukagoshi Y. Tsukagoshi Kitasato University, Sagamihara, Japan N. Shimizu N. Shimizu Kitasato University, Sagamihara, Japan M. IIzuka M. IIzuka Kitasato University, Sagamihara, Japan K. Eshima K. Eshima Kitasato University, Sagamihara, Japan N. Shinohara N. Shinohara Kitasato University, Sagamihara, Japan Abstract Signaling through TCR can result in a variety of distinct cellular responses in T cells. SIT is a transmembrane adaptor protein which is thought to play a regulatory role in TCR signaling. However, how SIT down-regulates the signaling cascades is largely unknown. In order to study the function of SIT, monoclonal antibodies have been developed. Flowcytometric analyses using these antibodies confirmed the expression of SIT on developing and mature lymphocytes. These observations were also confirmed by Western and Northern blotting. Analysis on DN, DP and SP subpopulations of thymocytes indicated that SIT is expressed on the cells of all four populations. SIT-expression starts at DN1 stage and reaches its peak around DN4-DP stages when developing cells undergo clonal selections through TCR-mediated signals. These observation are consistant with the nation that SIT plays a key role in clonal selections. Furthermore, SIT-protein was found also on B cells of very young (suckling) mice. Detailed analyses revealed that such SIT-expressing B cells in suckling mice had immature phenotypes resembling those found in bone marrow. Thus, the cells of the B lineage in adult bone marrow were examined and were shown to express SIT. These observations indicate that SIT is transiently expressed in cells of the B-lineage coinciding with rearrangement of Ig structural genes. SIT might play an important role in the Ig-receptor-mediated signal transduction in B cell development, as is the case with the cells of the T-lineage. PP-086-21 Subsets of human T follicular helper cells in health and autoimmunity R. Morita R. Morita 1 Keio University School of Medicine, Tokyo, Japan 2 Baylor Institute for Immunology Research, Dallas, TX, United States N. Schmitt N. Schmitt 2 Baylor Institute for Immunology Research, Dallas, TX, United States V. Pascual V. Pascual 2 Baylor Institute for Immunology Research, Dallas, TX, United States J. Banchereau J. Banchereau 2 Baylor Institute for Immunology Research, Dallas, TX, United States 3 INSERM U899, Dallas, TX, United States 4 Mount Sinai School of Medicine, New York, NY, United States H. Ueno H. Ueno 2 Baylor Institute for Immunology Research, Dallas, TX, United States 3 INSERM U899, Dallas, TX, United States Abstract T follicular helper cells (Tfh) cells, a lymph-resident CD4+ T cell subsetexpressing CXCR5, represent a T cell subset specialized for B cell help, and are considered to be distinct from Th1, Th2, and Th17 cell subsets. Over-representation of Tfh cells has been implicated in the pathogenesis of autoantibody-associated diseases in mouse models. CXCR5+CD4+ T cells also circulate in human blood, but little is known regarding their biological function and implication in autoimmune disease. We show here that blood CXCR5+CD4+ T cells represent a pool of Tfh cells, and are capable of helping naïve B cells to switch isotypes and differentiate into plasma cells. Based on cytokine secretion patterns and ability to help B cells, blood Tfh cells were found to be composed of subsets. Th2-committed CXCR3-CCR6- and Th17-committed CXCR3-CCR6+ cells can help naïve B cells through the secretion of IL-21, while Th1-committed CXCR3+CCR6- cells cannot. Patients with juvenile dermatomyositis, an autoantibody-associated disease, displayed a significant skewing of blood Tfh subsets towards Th2 and Th17. Significantly, the skewing of blood Tfh subsets correlated with disease activity and the frequency of blood plasmablasts. Collectively, our results demonstrate that human Tfh are composed of subsets, whose altered balance might contribute to autoimmunity. PP-086-22 Apoptosis and active inflammatory markers in chronic hepatitis C patients R. M. Issa R. M. Issa Institute of genetic engineering and biotechnology, Cairo, Egypt Abstract Aim of work;to assess the relationship between serum concentrations of adhesion and apoptotic -related soluble structures in patients affected by hepatitis virus C .Investigate serum levels of soluble Fas antigen (SFAS),soluble intracellular adhesion molecules(sICAM),study their role in pathogenesis of liver disease .Patients and methods, sixty chronic hepatitis C patients and twenty controls .They were classified into group A with minimal activity, group B with mild activity,group C with moderate and severe activity,depending on histological activity index score(HAI).Patients were classified into cirrhotic and non cirrhotic groups.Tissue Fas(tfas) was assessed usin anti fas antibody, sFas and ICAM using elisa .Result;sfas was significantly increased in patients subgroups compared to controls, and in comparing cirrhotic to non cirrhotic, tfas was rising with disease activity,in addition sfas corrolated with tfas except group A.Conclusion Fas studied parameters can reflect severity of liver inflammation and play a role in HCV infection © The Japanese Society for Immunology. 2010. 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TI - Systems immunology (PP-086)
JF - International Immunology
DO - 10.1093/intimm/dxq266
DA - 2010-08-01
UR - https://www.deepdyve.com/lp/oxford-university-press/systems-immunology-pp-086-8FvZVMttWU
SP - iv123
VL - 22
IS - Suppl_1_Pt_4
DP - DeepDyve
ER -