Abstract / Cytokine 63 (2013) 243–314 We have discovered a new recombinant type I interferon and named it as Super Interferon-I (sIFN-I) which showed signiﬁcant anti-virus and anti-tumor effect. Brieﬂy, the EC50 for HIV, sIFN-I was about 1000 fold lower than that IFNa-2b. For Ebola virus infected mice in the P4 lab of United States Department of Defense, 50% mice have been rescued by sIFN-I while 0% for regular IFNa-2b. For one lung cancer patient treated by sIFN-I alone, all the pleural ﬂuid can be completely eliminated and the tumor mass was reduced about 50%. For other lung cancer patients, by the combined treatment with sIFN-I and chemotherapy, the patients have got complete remissions. This sIFN-I is in clinical trial in Singapore and is applying for United States FDA clinical trial with good progression. We will also report the signiﬁcant molecular mechanism difference between sIFN-I and IFNa-2b, as explored in my lab during the past two years. http://dx.doi.org/10.1016/j.cyto.2013.06.052
rophages themselves are key regulators of this transition, and that the surface enzyme CD39 plays a critical role in self-limiting the activation process. Here, we reveal that toll-like receptor (TLR)-stimulated macrophages modulate their activation state by increasing the synthesis and secretion of adenosine triphosphate (ATP). We further demonstrate that macrophage-derived ATP is paradoxically immunosuppressive due to its rapid catabolism into adenosine by CD39. Macrophages lacking CD39 are unable to transition to a regulatory state and consequently continue to produce inﬂammatory cytokines. The importance of this transition was demonstrated in a mouse model of sepsis, where the presence of CD39-deﬁcient macrophages was sufﬁcient to induce lethal endotoxic shock. Thus, these data implicate CD39 as a key molecular switch that allows macrophages to intrinsically regulate their activation state. We propose that therapeutics targeting the release and hydrolysis of ATP by macrophages may represent new ways to treat inﬂammatory diseases. http://dx.doi.org/10.1016/j.cyto.2013.06.054
50 Evaluating inﬂuences of fructans on cytokine and immunoglobulin production in a human clinical trial Sandra T. Clarke a, Martin Kalmokoff b, Stephen P.J. Brooks c, Premysl Bercik d, Dan Ramdath e, Julia M. Green-Johnson a, a Applied Bioscience Graduate Program & Faculty of Science, University of Ontario Institute of Technology, Oshawa, Ont., Canada, b Atlantic Food and Horticulture Research Center, Agriculture and Agri-Food Canada, Kentville, NS, Canada, c Bureau of Nutritional Sciences, Health Canada, Ottawa, Ont., Canada, d Department of Medicine, McMaster University Medical Center, Hamilton, Ont., Canada, e Guelph Food Research Centre, Agriculture and Agri-Food Canada, Guelph, Ont., Canada A double-blind, placebo-controlled, randomized crossover clinical trial involving 30 healthy adults was conducted to examine the impact of fructans on both the gut microbiota and immune system. Along with their regular diet, trial participants consumed either an oligofructose-enriched inulin supplement as the test diet or food grade maltodextrin as the control diet for a 28 day period, with a minimum 2 week washout period before cross-over. The trial supplements were coded as ‘‘A” or ‘‘B”, and will remain coded until the completion of all analyses. Serum immunoglobulin concentrations were quantiﬁed by Luminex multiplexing technology. IgM was significantly higher after 28 days of supplement A consumption (821.3 ng/mL ± 138.3) than after supplement B consumption (485.8 ng/mL ± 66.5) (P = 0.0118). IgA was also signiﬁcantly higher (P = 0.0092) after supplement A consumption (1967.3 ng/ mL ± 374.4) than after supplement B consumption (911.7 ng/mL ± 138.2). Active transforming growth factor-beta (TGF-b) concentration was measured in plasma and serum cytokine concentrations were quantiﬁed by Enzyme-linked immunosorbent assay (ELISA). Levels of active TGF-b were 5.4 pg/mL ± 2.1 after consumption of supplement B, in comparison to 3.4 pg/mL ± 1.5 after supplement A consumption, although this difference was not statistically signiﬁcant. IL-10 concentrations were signiﬁcantly lower (P = 0.0172) after supplement B consumption (41.2 pg/mL ± 29.0) than after supplement A consumption (79.8 pg/mL ± 42.1). Granulocyte colony-stimulating factor (G-CSF) levels were also signiﬁcantly lower (P = 0.0265) after supplement B consumption (5.4 pg/mL ± 2.5) than levels after supplement A consumption (14.4 pg/mL ± 3.6). No differences were observed between subjects consuming supplement A and B in Interleukin-8 (IL-8) and IL-1Ra concentrations. Analysis of cytokine proﬁles will provide insight into the extent of the potential effects of dietary fructans on the immune system mediated through the gut microbiota.
51 Toll-like receptor stimulated macrophages intrinsically control inﬂammatory cytokine production via CD39-based mechanism Heather B. Cohen a,b, Katharine T. Briggs c, John P. Marino c,d, Katya Ravid e, Simon C. Robson f, David M. Mosser a,b, a University of Maryland, College Park Department of Cell Biology and Molecular Genetics, College Park, MD, USA, b Maryland Pathogen Research Institute, College Park, MD, USA, c Institute of Bioscience and Biotechnology Research, Rockville, MD, USA, d National Institute of Standards and Technology, Rockville, MD, USA, e Boston University, School of Medicine, Boston, MA, USA, f Harvard Medical School, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA, USA Sepsis is a highly fatal disease caused by an initial hyper-inﬂammatory response followed by a state of profound immunosuppression. Moreover, a coinciding switch in macrophage activation states has been observed in septic individuals over time. While, it is well-understood that the initial production of pro-inﬂammatory cytokines by macrophages accompanies the onset of sepsis, it remains unclear what drives the development of anti-inﬂammatory cytokines and the transition to an immunosuppressive state in macrophages. In this study, we demonstrate that mac-
52 Programmed cell death 4 (PDCD4) regulates proinﬂammatory cytokine signaling in bacterial pneumonia Taylor S. Cohen, Alice Prince, Columbia University, New York, NY, USA The balance between pro and anti-inﬂammatory signaling in innate immune responses to bacterial infection is especially critical in the lung. PDCD4, known primarily for its anti-tumor functions, inﬂuences inﬂammatory cytokine production and is negatively regulated by miR-21 (transcription) and p70S6K (phosphorylation), which are both targets of interferon (IFN) signaling. We postulated that PDCD4 has a central role in activating cytokine signaling, and functions as a target for IFNk. To conﬁrm the relationship between PDCD4 and inﬂammatory cytokines in the context of P. aeruginosa PAK infection, 16HBEs were treated with siRNA against PDCD4, stimulated with PAK (MOI 10, 4 h) and IL-8 was measured by RT-PCR. IL-8 induction was decreased in siRNA treated cells. Conversely, inhibiting miR-21 increased IL-8 induction in response to PAK. In response to PAK (MOI 10, 4 h), IFNk mRNA was increased >10-fold in dendritic cells. IFNk treatment of 16HBEs resulted in a STAT3 dependent decrease in miR-21 and increase in PDCD4 expression. IFNk also reduced phosphorylation of p70S6K. The in vivo relevance of these pathways was determined using wt and IL-28R-/- (IFNk receptor) mice. Similar levels of PDCD4 and miR-21 were found in the lungs of uninfected mice, however constitutive phosphorylation of p70S6K and PDCD4 was observed in knockout mice. IL-28R-/- mice had signiﬁcantly improved clearance of PAK from the lung (p < 0.05 for each) and improved lung pathology, correlating with decreased expression of KC and TNF and increased IL10 (p < 0.05), not immune cells (neutrophil, DC, macrophage) recruitment. Following 18 h of infection, miR-21 expression was signiﬁcantly higher (p = 0.0018) and PDCD4 mRNA signiﬁcantly lower (p = 0.0323) in IL-28R-/- compared to wt. Phosphorylation of p70S6K and PDCD4 in response to PAK was observed in WT and KO mice. These results indicate that PDCD4 is a central regulator of inﬂammatory cytokine production during P. aeruginosa pneumonia.
53 Inhibition of interleukin-15 dependent cytotoxic T-cell proliferation by the action of adenosine on dendritic cells Yair Cohen, Hadar Eini, Cidio Chaimovitz, Amos Douvdevani, Department of Clinical Biochemistry and Pharmacology, Soroka Medical Center and Ben-Gurion University of the Negev, Beer-Sheva, Israel Dendritic cells (DCs) regulate the immune response through production of various cytokines and signaling molecules, among which is IL-15, an important NK and cytotoxic T-cell (CTL) activator and survival factor. At the site of immune reaction, adenosine is produced from ATP by CD39 and CD73, ecto-enzymes expressed on regulatory T-cells (Treg). The aim of this present study was to examine the effect of adenosine on IL-15 production by DCs and the implication of this effect on CTL proliferation. Bone marrow derived DCs (BMDCs) were isolated from ICR mice. BMDCs were exposed to various adenosine receptor agonists/antagonist prior to stimulation with LPS and interferon-c (IFN-c). Levels of cAMP, IL-15 and IL-15 receptor a chain (IL15Ra) were measured using QPCR and/or ELISA. To assess the effect on CTL proliferation BMDCs were gamma-radiated and co-cultured with CTLL-2, a cytotoxic T-cell line, in the presence or absence of adenosine. Stimulation of BMDCs with LPS and IFN-c increased IL-15 and IL-15Ra levels. Treatment of stimulated BMDCs with adenosine signiﬁcantly reduced IL-15/IL-15Ra mRNA and protein levels. This down-regu-