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Introduction Approximately 15% of appropriately treated patients with early syphilis remain serofast. The pathogenesis and clinical significance of this phenomenon are unclear. Aim To determine the significance of Treponema pallidum-specific immune responses and autoimmunity in the treatment outcome of syphilis (serofast or proper serological response). Material and methods Forty-eight patients with secondary and early latent syphilis (ELS) were enrolled in this study. Reactivity of IgM/IgG antibodies to the treponemal antigens TpN47, TpN17, TpN15 and TmpA was evaluated before and 12 months after intramuscular penicillin therapy for syphilis. Additionally, the presence of antinuclear antibodies (ANA) was determined 12 months after treatment. Results After 1 year, patients were stratified into two groups based on their serological response: (1) serofast (n = 10) and (2) serologically-cured (n = 38) patients. The serological cure rate was 79.2% at 12 months after treatment. Weak pre- and post-treatment antibody reactivity to TpN47 antigen was found to be significantly associated with a higher risk of the serofast state (OR = 64; 95% CI: 5.01-817; p < 0.005). Patients who remained serofast had a significantly higher ANA prevalence and mean titer when compared to those with proper serological responses (100% vs. 5.3%, respectively, p < 0.005; 1 : 640 vs. 1 : 160, respectively, p < 0.005). Conclusions We demonstrate that baseline antigen-specific immune response to Treponema pallidum may be an important predictor of the treatment outcome. Further studies are warranted to identify the role of autoimmunity in the pathomechanism of the serofast state.

Background OGN could modify tissue inflammation and immune response via local and circulating innate immune cells, which was suggestive of a reciprocal relationship between OGN and T cell infiltration in cancer. Hence, we aim to measure the OGN expression patterns and immune cells response in colorectal cancer(CRC). Methods This study enrolled three independent sets of patients from TCGA and the Fudan University Shanghai Cancer Center(FUSCC). The effect of OGN on T cell infiltration and the mechanism were examined in vitro and in vivo. Findings Tumor OGN expression levels were positively associated with CD3, CD8, and PTPRC expressions in the training and testing sets from TCGA, respectively. In validation set from FUSCC, OGN expression level also paralleled positively with CD8+ cell density in colorectal cancer tissue (p, Highlights • OGN is positively associated with T cell density in CRC • OGN inhibited the activation of HIF-1α, then significantly impeded the expression of VEGF.

External beam radiotherapy (EBRT) with carbon ions and endoradiotherapy using radiolabeled tumor targeting agents are emerging concepts in precision cancer therapy. We report on combination effects of these two promising strategies. Tumor targeting 131I-labelled anti-EGFR-antibody (Cetuximab) was used in the prototypic EGFR-expressing A431 human squamous cell carcinoma xenograft model. A 131I-labelled melanin-binding benzamide derivative was utilized targeting B16F10 melanoma in an orthotopic syngeneic C57bl6 model. Fractionated EBRT was performed using carbon ions in direct comparison with conventional photon irradiation. Tumor uptake of 131I-Cetuximab and 131I-Benzamide was enhanced by fractionated EBRT as determined by biodistribution studies. This effect was independent of radiation quality and significant for the small molecule 131I-Benzamide, i.e., >30% more uptake in irradiated vs. non-irradiated melanoma was found (p

A wheat LLM-domain-containing B-GATA transcription factor positively regulates the host immune response to the important pathogen Rhizoctonia cerealis., Wheat (Triticum aestivum) is essential for global food security. Rhizoctonia cerealis is the causal pathogen of sharp eyespot, an important disease of wheat. GATA proteins in model plants have been implicated in growth and development; however, little is known about their roles in immunity. Here, we report a defence role for a wheat LLM-domain-containing B-GATA transcription factor, TaGATA1, against R. cerealis infection and explore the underlying mechanism. Through transcriptomic analysis, TaGATA1 was identified to be more highly expressed in resistant wheat genotypes than in susceptible wheat genotypes. TaGATA1 was located on chromosome 3B and had two homoeologous genes on chromosomes 3A and 3D. TaGATA1 was found to be localized in the nucleus, possessed transcriptional activation activity, and bound to GATA-core cis-elements. TaGATA1 overexpression significantly enhanced resistance of transgenic wheat to R. cerealis, whereas silencing of TaGATA1 suppressed the resistance. Quantitative reverse transcription–PCR and ChIP–qPCR results indicated that TaGATA1 directly bound to and activated certain defence genes in host immune response to R. cerealis. Collectively, TaGATA1 positively regulates immune responses to R. cerealis through activating expression of defence genes in wheat. This study reveals a new function of plant GATAs in immunity and provides a candidate gene for improving crop resistance to R. cerealis.

Anti-microbial peptides are produced at outer and inner surfaces by epithelia and innate immune cells in response to bacterial infection. Staphylococcus aureus is an enterotoxin producing, Gram-positive pathogen, which is a major cause of soft tissue infections and life-threatening bacteremia and sepsis. Here we show that (i) skin T cells in chronic wounds infected with S. aureus express interleukin-26 (IL-26) in situ, (ii) staphylococcal enterotoxins (SE) trigger IL-26 expression in T cell lines and primary skin T cells, and (iii) IL-26 triggers death and inhibits biofilm formation and growth of S. aureus. Thus, we provide novel evidence that IL-26 is an anti-microbial peptide produced by T cells in response to SE. Accordingly, we propose that IL-26 producing T cells take part in the innate immune response to SE producing S. aureus and thus play a novel role in the primary innate immune defense in addition to their classical role in adaptive immunity. Published version

The composition of the intestinal microbiota is related to the health and immune function of the host. Administration of antibiotics affects the composition of the intestinal microbiota. However, the effects of immune function on the composition of the intestinal microbiota are still unclear. In this study, we investigated the lymphocyte composition and determined the relationships between lymphocyte function and the intestinal microbiota following antibiotic treatment in mice. To change the composition of the intestinal microbiota, mice were treated with or without antibiotics. Analysis of intestinal microbiota was performed by metagenomic analysis targeting 16S rRNA. Lymphocyte subsets of splenocytes were measured by flow cytometry. For functional analysis of T cells, splenocytes were stimulated with concanavalin (Con A), and cytokine gene expression was measured by real-time polymerase chain reaction. Firmicutes were predominant in the control group, whereas Bacteroidetes predominated in the antibiotic-treated group, as determined by metagenomic analysis. The diversity of the microbiota decreased in the antibiotic-treated group. Analysis of lymphocyte subsets showed that CD3(+) cells decreased, whereas CD19(+) cells increased in the antibiotic-treated group. All cytokine genes in splenocytes treated with Con A were downregulated in the antibiotic-treated group; in particular, genes encoding interferon-gamma, interleukin (IL)-6, and IL-13 significantly decreased. Taken together, these results revealed that changes in the composition of the intestinal microbiota by antibiotic treatment influenced the population of lymphocytes in splenocytes and affected the immune response.

Depending on the length of their carbon backbone and their saturation status, natural fatty acids have rather distinct biological effects. Thus, longevity of model organisms is increased by extra supply of the most abundant natural cis-unsaturated fatty acid, oleic acid, but not by that of the most abundant saturated fatty acid, palmitic acid. Here, we systematically compared the capacity of different saturated, cis-unsaturated and alien (industrial or ruminant) trans-unsaturated fatty acids to provoke cellular stress in vitro, on cultured human cells expressing a battery of distinct biosensors that detect signs of autophagy, Golgi stress and the unfolded protein response. In contrast to cis-unsaturated fatty acids, trans-unsaturated fatty acids failed to stimulate signs of autophagy including the formation of GFP-LC3B-positive puncta, production of phosphatidylinositol-3-phosphate, and activation of the transcription factor TFEB. When combined effects were assessed, several trans-unsaturated fatty acids including elaidic acid (the trans-isomer of oleate), linoelaidic acid, trans-vaccenic acid and palmitelaidic acid, were highly efficient in suppressing autophagy and endoplasmic reticulum stress induced by palmitic, but not by oleic acid. Elaidic acid also inhibited autophagy induction by palmitic acid in vivo, in mouse livers and hearts. We conclude that the well-established, though mechanistically enigmatic toxicity of trans-unsaturated fatty acids may reside in their capacity to abolish cytoprotective stress responses induced by saturated fatty acids., Graphical Abstract Unlabelled Image, Highlights • The molecular geometry of fatty acids dictates their biological function • Saturated fats induce autophagy through a BECN1-dependent pathway • Cis fats trigger cytoprotection via the induction of non-canonical autophagy, trans fats are toxic as they inhibit the stress response induced by saturated fats Cellular responses to fatty acids are distinct according to their unsaturation status and carbon backbone; saturated & cis fats can induce various cellular stresses, including autophagy (through a BECN1-dependent pathway for the first category only), whereas trans fats are toxic by specifically inhibiting saturated fats-elicited stress responses.

// Consuelo Amantini 1,* , Valerio Farfariello 2,* , Claudio Cardinali 3,4 , Maria Beatrice Morelli 3,4 , Oliviero Marinelli 1 , Massimo Nabissi 3 , Matteo Santoni 3 , Laura Bonfili 1 , Valentina Cecarini 1 , Anna Maria Eleuteri 1 and Giorgio Santoni 3 1 School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy 2 University of Lille, INSERM U1003 - PHYCEL - Physiologie Cellulaire, Lille, France 3 School of Pharmacy, Experimental Medicine Section, University of Camerino, Camerino, Italy 4 Department of Molecular Medicine, Sapienza University, Rome, Italy * These authors have contributed equally to this work Correspondence to: Consuelo Amantini, email: // Keywords : ER stress, capsaicin, TRPV1, TRPV1 KO mice, autophagy, Immunology and Microbiology Section, Immune response, Immunity Received : July 28, 2017 Accepted : September 20, 2017 Published : October 11, 2017 Abstract Autophagy and the ubiquitin-proteasome system (UPS) control thymus cell homeostasis under resting and endoplasmic reticulum (ER) stress conditions. Several evidence support a cross-talk between UPS and autophagy; abrogation of UPS responses stimulates autophagy, and vice versa the inhibition of autophagy alters the UPS functions. Herein, we found that TRPV1 activation induces ER stress, proteasome dysfunction and autophagy in thymocytes by modulating the expression of UPR-related genes. The TRPV1-mediated autophagy prevents the UPR activation by inhibiting BiP, Grp94 and ERp57 chaperone protein expression. Thymocytes from TRPV1 KO mice display both autophagy and proteasome dysfunctions, resulting in increased apoptotic cells and reduced total DP thymocyte number. In addition, positive selection of thymocytes triggered by anti-TCRβ/CD2 Ab-mediated costimulation induces apoptosis in thymocytes from TRPV1 KO as compared with WT mice. Stimulation of TRPV1 KO thymocytes with anti-TCRβ/CD2 mAbs modulates the expression of CD4 antigen on purified DP thymocytes, with reduced number of mature, single positive (SP) CD4 and increased number of immature SP CD4 low and DP CD4 low CD8 + thymocytes, further supporting the intrinsic role of TRPV1 in T cell maturation. Finally, a reduction in CD8 + and CD4 + T cells is evidenced in the peripheral blood and spleen of TRPV1 KO, as compared with WT mice. Therapeutic strategy by restraining or stimulating the TRPV1 expression and functions in thymocytes might represent a new pharmacological tool in the regulation of different inflammatory T cell responses.

Amphioxus is the closest living proxy for exploring the evolutionary origin of the immune system in vertebrates. To understand the immune responses of amphioxus to lipopolysaccharide (LPS), 5 ribosomal RNA (rRNA)-depleted libraries of amphioxus were constructed, including one control (0 h) library and 4 treatment libraries at 6, 12, 24, and 48 h post-injection (hpi) with LPS. The transcriptome of Branchiostoma belcheri was analyzed using strand-specific RNA sequencing technology (RNA-seq). A total of 6161, 6665, 7969, and 6447 differentially expressed genes (DEGs) were detected at 6, 12, 24, and 48 hpi, respectively, compared with expression levels at 0 h. We identified amphioxus genes active during the acute-phase response to LPS at different time points after stimulation. Moreover, to better visualize the resolution phase of the immune process during immune response, we identified 6057 and 5235 DEGs at 48 hpi by comparing with 6 and 24 hpi, respectively. Through real-time quantitative PCR (qRT-PCR) analysis of 12 selected DEGs, we demonstrated the accuracy of the RNA-seq data in this study. Functional enrichment analysis of DEGs demonstrated that most terms were related to defense and immune responses, disease and infection, cell apoptosis, and metabolism and catalysis. Subsequently, we identified 1330, 485, 670, 911, and 1624 time-specific genes (TSGs) at 0, 6, 12, 24, and 48 hpi. Time-specific terms at each of 5 time points were primarily involved in development, immune signaling, signal transduction, DNA repair and stability, and metabolism and catalysis, respectively. As this is the first study to report the transcriptome of an organism with primitive immunity following LPS challenge at multiple time points, it provides gene expression information for further research into the evolution of immunity in vertebrates.

Anti-tumor necrosis factor (TNF) therapies are successful in the treatment of inflammatory disorders. However, some patients with rheumatoid arthritis (RA) fail to response anti-TNF drugs due to the compensation of other inflammatory signals. In this study, to reduce compensatory responses of interleukin-17A (IL-17A) during TNF-α inhibition, we generated an IgG-like bispecific antibodiy (bsAb) against TNF-α and IL-17A through a combination method of electrostatic Fc pairing and light chain crossover. This bsAb exhibited relatively high stability comparable to natural IgG antibodies, and retained the unaltered affinities to both of two targets. BsAb significantly decreased not only the expression level of neutrophil or Th17 chemokines, but also the secretion of IL-6/IL-8 on fibroblast-like synoviocytes (FLS) from a patient with RA. Meanwhile, TNF-α-mediated cellular cytotoxicity of fibroblasts was neutralized by bsAb. Importantly, we demonstrate that the combined blockade of TNF-α and IL-17A is more efficient than inhibition of either factor alone. Our results suggest the IgG-like anti-TNF-α/IL-17A bispecific molecule overcome the limited therapeutic responses using anti-TNF drugs. It may be a promising therapeutic agent for the treatment of autoimmune diseases.

// Masahiko Ito 1 , Suofeng Sun 1 , Takasuke Fukuhara 2 , Ryosuke Suzuki 3 , Miho Tamai 4 , Toyohiko Yamauchi 5 , Kenji Nakashima 1 , Yoh-ichi Tagawa 4 , Shigetoshi Okazaki 6 , Yoshiharu Matsuura 2 , Takaji Wakita 3 and Tetsuro Suzuki 1 1 Department of Virology and Parasitology, Hamamatsu University School of Medicine, Shizuoka, Japan 2 Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan 3 Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan 4 School of Life Science and Technology, Tokyo Institute of Technology, Kanagawa, Japan 5 Central Research Laboratory, Hamamatsu Photonics K.K., Shizuoka, Japan 6 Department of Medical Spectroscopy, Hamamatsu University School of Medicine, Shizuoka, Japan Correspondence to: Tetsuro Suzuki, email: // Keywords : hepatitis C virus, epigenetic reprogramming, hepatoma cell, oval cell, miR200a, Immunology and Microbiology Section, Immune response, Immunity Received : May 16, 2017 Accepted : June 28, 2017 Published : July 08, 2017 Abstract Directed differentiation of human stem cells including induced pluripotent stem cells into hepatic cells potentially leads to acquired susceptibility to hepatitis C virus (HCV) infection. However, cellular determinants that change their expression during cell reprogramming or hepatic differentiation and are pivotal for supporting the HCV life cycle remain unclear. In this study, by introducing a set of reprogramming factors, we established HuH-7-derived oval-like cell lines, Hdo-17 and -23, which possess features of bipotential liver precursors. Upon induction of hepatocyte differentiation, expression of mature hepatocyte markers and hepatoblast markers in cells increased and decreased, respectively. In contrast, in response to cholangiocytic differentiation induction, gene expression of epithelium markers increased and cells formed round cysts with a central luminal space. Hdo cells lost their susceptibility to HCV infection and viral RNA replication. Hepatic differentiation of Hdo cells potentially led to recovery of permissiveness to HCV RNA replication. Gene expression profiling showed that most host-cell factors known to be involved in the HCV life cycle, except CD81, are expressed in Hdo cells comparable to HuH-7 cells. HCV pseudoparticle infectivity was significantly but partially recovered by ectopic expression of CD81, suggesting possible involvement of additional unidentified factors in HCV entry. In addition, we identified miR200a-3p, which is highly expressed in Hdo cells and stem cells but poorly expressed in differentiated cells and mature hepatocytes, as a novel negative regulator of HCV replication. In conclusion, our results showed that epigenetic reprogramming of human hepatoma cells potentially changes their permissivity to HCV.

Excretory/secretory antigens (ESAs) produced by Toxoplasma gondii enable this parasite to invade and survive within the host cells through immunomodulation. In this study, the modulating effects of T. gondii excretory/secretory antigens (TgESAs) on the Ana-1 murine macrophage cell line were evaluated. Ana-1 cells were incubated with several concentrations of TgESAs, and the resulting effects on cellular viability, phagocytotic ability, and apoptosis induction were determined. Pro-inflammatory and anti-inflammatory cytokine secretion, nitric oxide production, toll-like receptor expression, and nuclear translocation of NF-κB were all measured after incubation with TgESAs. After TgESAs treatment, the proliferation and phagocytosis ability of Ana-1 cells decreased, and apoptosis was induced in a dose dependent manner. Analysis of Ana-1 cell culture supernatants showed that TgESAs not only upregulated secretion of anti-inflammatory cytokines (interleukin-10 and transforming growth factor-β1), they also inhibited secretion of pro-inflammatory cytokines (tumor necrosis factor-α and interleukin-1β). Additionally, TgESAs inhibited nitric oxide production, toll-like receptor (TLR) 2 and 4 activation, and the nuclear translocation of NF-κB in lipopolysaccharide-stimulated Ana-1 macrophages. These results suggest TgESAs inhibit the functional activity of Ana-1 murine macrophages by inhibiting TLR-induced NF-κB activation, which suppresses pro-inflammatory cytokine secretion.

// Ying Lu 1 , Xianyu Zhang 1 , Nicolas Bouladoux 2 , Saransh Neel Kaul 3 , Kangxin Jin 4 , Derek Sant’Angelo 5 , Yasmine Belkaid 2 and Damian Kovalovsky 1,6 1 Experimental Immunology Branch, NCI, NIH, Bethesda, MD, USA 2 Mucosal Immunology Section, Laboratory of Parasitic Diseases, NIAID, NIH, Bethesda, MD, USA 3 University of Maryland, College Park, MD, USA 4 Zhongshan Ophthalmic Center, State Key Laboratory for Ophthalmic Researches, Sun Yat-Sen University, Guangzhou, Guangdong, China 5 Cancer Metabolism and Growth Program, Rutgers, Child Health Institute of New Jersey, New Brunswick, NJ, USA 6 Experimental Transplantation and Immunology Branch, NCI, NIH, Bethesda, MD, USA Correspondence to: Damian Kovalovsky, email: // Keywords : innate lymphoid cells, ILC, ILC3, Zbtb1, IFN-γ, Immunology and Microbiology Section, Immune response, Immunity Received : May 16, 2017 Accepted : July 14, 2017 Published : July 27, 2017 Abstract Innate lymphoid cells (ILCs) play a central role conferring protection at the mucosal frontier. In this study, we have identified a requirement of the transcription factor Zbtb1 for the development of RORγt + ILCs (ILC3s). Zbtb1-deficient mice lacked NKp46 + ILC3 cells in the lamina propria of the small and large intestine. This requirement of Zbtb1 was cell intrinsic, as NKp46 + ILC3s were not generated from Zbtb1-deficient progenitors in bone marrow chimeras and Zbtb1-deficient RORγt + CCR6 - NKp46 - ILC3s didn’t generate NKp46 + ILC3s in co-cultures with OP9-DL1 stroma. In correlation with this impairment, Zbtb1-deficient ILC3 cells failed to upregulate T-bet expression, and to acquire IFN-γ production characteristic of NKp46 + cells. Finally, absence of NKp46 + ILC3 cells combined with the absence of T-cells in Zbtb1-deficient mice, led to a transient susceptibility to C. rodentium infections. Altogether, these results establish that Zbtb1 is essential for the development of NKp46 + ILC3 cells.

// Ho-Chang Kuo 1,2 , Sung-Chou Li 3 , Lien-Hung Huang 3 and Ying-Hsien Huang 1,2,4 1 Department of Pediatrics, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan 2 Kawasaki Disease Center, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan 3 Department of Medical Research, Genomics and Proteomics Core Laboratory, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan 4 Department of Pediatrics, Chiayi Chang Gung Memorial Hospital, Kaohsiung, Taiwan Correspondence to: Ying-Hsien Huang, email: // Keywords : matrix metalloproteinase, genetic methylation, Kawasaki disease, Immunology and Microbiology Section, Immune response, Immunity Received : April 05, 2017 Accepted : June 28, 2017 Published : July 28, 2017 Abstract Background: Kawasaki disease (KD) is a type of febrile coronary vasculitis occurring in children. Some researchers have suggested that changes in genetic signatures, such as matrix metalloproteinases (MMPs), are critical markers for cardiovascular diseases. This study aims to provide a comprehensive survey of global DNA methylation levels and MMP transcripts of KD patients compared to control subjects. Materials and Methods: For chips studies, we recruited a total of 18 KD patients, prior to receiving intravenous immunoglobulin (IVIG) and at least 3 weeks after IVIG treatment, as well as 18 healthy and 18 febrile control subjects. We applied Illumina HumanMethylation450 BeadChip and Affymetrix GeneChip® Human Transcriptome Array 2.0 to evaluate their CpG markers and expression levels, respectively. Then we used a separate cohort to carry out real-time quantitative PCR validations of mRNA levels. Results: The expressions of mRNA levels of MMP-8, -9, and -25 were significantly upregulated in KD patients compared to the healthy and febrile controls. Once KD patients underwent IVIG treatment, these MMPs considerably decreased. In particular, the methylation status of CpG sites of MMP-9 indicated a significant opposite tendency between both stages of not only the KD samples but also the controls. We also observed the mRNA level of MMP-9 to be higher in KD patients with coronary arterial lesion formation. Conclusion: This study is the first to report epigenetic hypomethylation, an increased MMP-9 transcript, and the upregulation of MMP-9 in KD patients who had formed coronary arterial lesions.

// Jun Lou 1,2,* , Xiawei Li 1,2,* , Wei Huang 1,2 , Jingjing Liang 1,2 , Mingzhu Zheng 1,2 , Ting Xu 1,2 , Jun Lyu 1,2 , Dan Li 1,2 , Qin Xu 1,2 , Xuexiao Jin 1,2 , Guotong Fu 1,2 , Di Wang 1,2 and Linrong Lu 1,2 1 Institute of Immunology, Zhejiang University School of Medicine, Hangzhou, China 2 Program in Molecular and Cellular Biology, Zhejiang University School of Medicine, Hangzhou, China * These authors have equally contributed to this work Correspondence to: Linrong Lu, email: // Keywords : SNX10, macrophage, Listeria monocytogenes, phagosome, innate immunity, Immunology and Microbiology Section, Immune response, Immunity Received : May 16, 2017 Accepted : July 13, 2017 Published : July 27, 2017 Abstract Listeria monocytogenes ( L. monocytogenes ),which is a facultative intracellular bacterial pathogen that causes listeriosis, is widely used to study the mammalian immune response to infection. After phagocytosis by professional phagocytes, L. monocytogenes is initially contained within phagosomes, which mature into phagolysosomes, where the bacteria are degraded. Although phagocytosis and subsequent phagosome maturation is essential for the clearance of infectious microbial pathogens, the underlying regulatory mechanisms are still unclear. SNX10 (Sorting nexin 10) has the simplest structure of the SNX family and has been reported to regulate endosomal morphology, which might be crucial for macrophage function, including phagocytosis and digestion of pathogens, inflammatory response, and antigen presentation. Our results showed that SNX10 expression was upregulated following L. monocytogenes infection in macrophages. It was also revealed that SNX10 promoted phagosome maturation by recruiting the Mon1-Ccz1 complex to endosomes and phagosomes. As a result, SNX10 deficiency decreased the bacterial killing ability of macrophages, and SNX10-deficient mice showed increased susceptibility to L. monocytogenes infection in vivo . Thus, this study revealed an essential role of SNX10 in controlling bacterial infection.

// Jia Yi Guo 1,* , Feng Li 1,* , Yong Bing Wen 1 , Hong Xun Cui 1 , Ma Long Guo 1 , Lin Zhang 2 , Yun Fei Zhang 1 , Yan Jin Guo 1 and Yan Xing Guo 1 1 Luoyang Orthopedic Hospital of Henan Province, Luoyang, Henan, China 2 Department of Surgery, Advanced Clinical Skills Centre, University of Auckland, Auckland, New Zealand * These authors have contributed equally to this work Correspondence to: Hong-Xun Cui, email: // Yan Xing Guo, email: // Keywords : melatonin, redox, Sirt1, NAMPT, NFAT5, Immunology and Microbiology Section, Immune response, Immunity Received : December 19, 2016 Accepted : May 12, 2017 Published : June 03, 2017 Abstract Osteoarthritis (OA) is a degenerative joint disease mainly characterized by cartilage degradation. Interleukin-1β (IL-1β) contributes to OA pathogenesis by enhancing oxidative stress and inflammation. Melatonin reportedly elicits potent protection against OA. However, the role of melatonin and underlying mechanism in IL-1β-stimulated chondrocytes remain largely unclear. In this study, we found that melatonin inhibited IL-1β-induced toxicity and sirtuin 1 (Sirt1) enhancement in human chondrocytes. Melatonin reduced the IL-1β-increased nicotinamide phosphoribosyltransferase (NAMPT) expression and the NAD + level in chondrocytes in a Sirt1-dependent manner. In turn, the inhibitory effect of melatonin on Sirt1 was mediated by NAMPT. Moreover, melatonin suppressed IL-1β-induced Sirt1-mediated matrix metalloproteinase (MMP)-3 and MMP-13 production. Melatonin also decreased the Sirt1-steered nuclear factor of activated T cells 5 (NFAT5) expression in IL-1β-challenged chondrocytes. NFAT5 depletion mimicked the suppressive effects of melatonin on IL-1β-elevated production of inflammatory mediators, including tumor necrosis factor-α (TNF-α), IL-1β, prostaglandin E2 (PGE 2 ), and nitric oxide (NO) in chondrocytes. TNF-α, IL-1β, PGE 2 , or NO decrease caused the similar reduction of MMP-3 and MMP-13 by melatonin in IL-1β-insulted chondrocytes. Highly consistent with in vitro findings, in vivo results demonstrated that melatonin repressed the expression of relevant genes in rat OA pathogenesis in anterior cruciate ligament transection model. Overall, these results indicate that melatonin effectively reduced IL-1β-induced MMP production by inhibiting Sirt1-dependent NAMPT and NFAT5 signaling in chondrocytes, suggesting melatonin as a potential therapeutic alternative for chondroprotection of OA patients.

// Arcangelo Liso 1 , Stefano Castellani 1 , Francesca Massenzio 1 , Rosa Trotta 1 , Alessandra Pucciarini 2 , Barbara Bigerna 2 , Pasquale De Luca 3 , Pietro Zoppoli 4 , Filippo Castiglione 5 , Maria Concetta Palumbo 5 , Fabrizio Stracci 6 , Matteo Landriscina 1,7 , Giorgina Specchia 8 , Leon A. Bach 9,10 , Massimo Conese 1 and Brunangelo Falini 2 1 Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy 2 Institute of Haematology, University of Perugia, Perugia, Italy 3 Stazione Zoologica A. Dohrn, Naples, Italy 4 Dipartimento di Medicina Sperimentale e Clinica, Universita degli Studi Magna Graecia, Catanzaro, Italy 5 Institute for Applied Computing, National Research Council of Italy, Rome, Italy 6 Department of Experimental Medicine, Section of Public Health, University of Perugia, Perugia, Italy 7 Laboratory of Preclinical and Translational Research, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture, Italy 8 Institute of Hematology, University of Bari, Bari, Italy 9 Department of Medicine, Alfred Hospital, Monash University, Melbourne, Australia 10 Department of Endocrinology and Diabetes, Alfred Hospital, Melbourne, Australia Correspondence to: Arcangelo Liso, email: // Massimo Conese, email: // Keywords : apoptosis, B cells, chemotaxis, dendritic cells, hyperthermia, Immunology and Microbiology Section, Immune response, Immunity Received : April 16, 2017 Accepted : May 12, 2017 Published : June 01, 2017 Abstract Fever plays a role in activating innate immunity while its relevance in activating adaptive immunity is less clear. Even brief exposure to elevated temperatures significantly impacts on the immunostimulatory capacity of dendritic cells (DCs), but the consequences on immune response remain unclear. To address this issue, we analyzed the gene expression profiles of normal human monocyte-derived DCs from nine healthy adults subjected either to fever-like thermal conditions (39°C) or to normal temperature (37°C) for 180 minutes. Exposure of DCs to 39°C caused upregulation of 43 genes and downregulation of 24 genes. Functionally, the up/downregulated genes are involved in post-translational modification, protein folding, cell death and survival, and cellular movement. Notably, when compared to monocytes, DCs differentially upregulated transcription of the secreted protein IGFBP-6, not previously known to be specifically linked to hyperthermia. Exposure of DCs to 39°C induced apoptosis/necrosis and resulted in accumulation of IGFBP-6 in the conditioned medium at 48 h. IGFBP-6 may have a functional role in the hyperthermic response as it induced chemotaxis of monocytes and T lymphocytes, but not of B lymphocytes. Thus, temperature regulates complex biological DC functions that most likely contribute to their ability to induce an efficient adaptive immune response.

// Marcell Costa de Medeiros 1 , Rajat Banerjee 2 , Min Liu 2 , Giovana Anovazzi 1 , Nisha J. D’Silva 2,3 and Carlos Rossa Junior 1 1 Department of Diagnosis and Surgery, School of Dentistry at Araraquara, Sao Paulo State University, Araraquara, SP, Brazil 2 Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, Michigan, USA 3 Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, USA Correspondence to: Carlos Rossa Junior, email: // Nisha J. D’Silva, email: // Keywords : head and neck squamous cell carcinoma, immune response, T cell, immunosuppression and tumor scape, Immunology and Microbiology Section, Immune response, Immunity Received : January 12, 2017 Accepted : June 01, 2017 Published : June 15, 2017 Abstract The immune system detects shifts from homeostasis and eliminates altered cells. However, neoplastic cells can modulate the host response to escape immunosurveillance thereby allowing tumor progression. Head and neck squamous cell carcinoma (HNSCC) is one of the most immunosuppressive cancers but its role in co-opting the immune system to actively promote tumor growth has not been investigated. In this study, we investigated the influence of soluble factors secreted by HNSCC and non-neoplastic epithelial cells on proliferation, apoptosis, activation, cytokine gene expression and phenotypic polarization of immune cells of healthy donors. Then, we determined if the immunomodulation caused by HNSCC-derived soluble products leads to immunosubversion by assessing proliferation, migration and survival of tumor cells exposed to soluble products secreted by modulated immune cells or co-cultured with immune cells. Soluble products from HNSCC inhibited proliferation and cytokine expression in PBMCs, activation of T cells, and polarization of CD4+ towards the Th17 phenotype. These changes co-opted the immune cells to favor cell proliferation, survival and migration of HNSCC. This immunosubversion was observed both indirectly with secreted products and with direct cell-to-cell contact. We conclude that HNSCC-derived secreted products create an immunosuppressive environment that facilitates evasion of tumor cells and subverts the immune cells into a pro-tumoral phenotype.

// Qin Luo 1 , Hengmin Cui 1,2 , Huidan Deng 1 , Ping Kuang 1 , Huan Liu 1 , Yujiao Lu 1 , Jing Fang 1,2 , Zhicai Zuo 1,2 , Junliang Deng 1,2 , Yinglun Li 1,2 , Xun Wang 1,2 and Ling Zhao 1,2 1 College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, China 2 Key Laboratory of Animal Diseases and Environmental Hazards of Sichuan Province, Sichuan Agriculture University, Wenjiang, Chengdu, China Correspondence to: Hengmin Cui, email: // Keywords : sodium fluoride, oxidative damage, lesion, dysfunction, kidney, Immunology and Microbiology Section, Immune response, Immunity Received : March 13, 2017 Accepted : April 11, 2017 Published : April 21, 2017 Abstract It has been reported that excessive intake of fluoride can induce renal lesions. However, its pathogenesis is still less understood. Therefore, this study was conducted to investigate oxidative damage and the relationships between the oxidative damage and renal lesions in fluoride-treated mice by using the methods of histopathology, biochemistry, flow cytometry and quantitative real-time polymerase chain reaction (qRT-PCR). A total of 240 ICR mice were randomly divided into four equal groups (sodium fluoride was given orally at the dose of 0, 12, 24 and 48 mg/kg body weight for 42 days, respectively). We found that fluoride in excess of 12 mg/kg induced renal oxidative damage, which was characterized by increasing the levels of reactive oxygen species (ROS) production and contents of malondialdehyde (MDA) and protein carbonyls (PC), and decreasing the abilities of anti-superoxide anion (ASA) and anti-hydroxyl radical (AHR), glutathione (GSH) content, as well as activities and mRNA expression levels of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GSH-Px). Concurrently, fluoride caused degeneration and necrosis of the tubular cells, renal tubular hyaline casts and glomeruli swelling, which were consistent with the alteration of renal function parameters including elevated contents of serum creatinine (Cr), serum uric acid (UA), blood urea nitrogen (BUN), and the activities of urinary N-acetyl-b-D-glucosaminidase (NAG), renal lactate dehydrogenase (LDH), and reduced activities of sodium-potassium adenosine triphosphatase (Na+/K+-ATPase) and acid phosphatase (ACP) in the kidney. The above-mentioned results showed that fluoride in excess of 12 mg/kg induced renal oxidative damage, which then caused renal lesions and dysfunctions. These findings also clearly demonstrated that oxidative damage is one of the mechanisms of fluoride-induced renal lesions and dysfunctions.

// Lijie Sun 1,2,* , Yan Li 1,2,* , Xiuzhi Jia 1,2,* , Qi Wang 1,2 , Yue Li 3 , Minghui Hu 4 , Linlu Tian 1,2 , Jinfeng Yang 1,2 , Wenjing Xing 1,2 , Weihua Zhang 5 , Jingtao Wang 6 , Hongwei Xu 1,2 , Lihua Wang 7 , Dekai Zhang 3 and Huan Ren 1,2 1 Department of Immunology, Harbin Medical University, Harbin, China 2 Key Laboratory of Infection & Immunity, Heilongjiang Province, Harbin, China 3 Center for Infectious and Inflammatory Disease, Institute of Biosciences and Technology, Texas A&M University System Health Science Center, Houston, TX, USA 4 Department of Clinical Laboratory, The Affiliated Hospital to Qingdao University, Qingdao, China 5 Department of Pathophysiology, Harbin Medical University, Harbin, China 6 Department of Epidemiology and Biostatistics, The Public Health Institute, Harbin Medical University, Harbin, China 7 Department of Neuroscience, The Second Hospital Affiliated to Harbin Medical University, Harbin, China Correspondence to: Huan Ren, email: // Dekai Zhang, email: // Lihua Wang, email: // Keywords : interferon-gamma, astrocyte, interleukin-6, apoptosis, neuroinflammation, Immunology and Microbiology Section, Immune response, Immunity Received : November 21, 2016 Accepted : March 22, 2017 Published : April 09, 2017 Abstract Inflammation eliminates pathogenic infections while also threatening the integrity of the central nervous system. In this study, using in vivo and in vitro models of acute neuroinflammation, we investigated the mechanisms by which inflammation and astrocytes affect neuronal apoptosis. The in vitro model mimicked acute neuroinflammation by incubation in IFN-γ-containing media with primary cultured cerebellar granule neurons, with or without cultured astrocytes. This quickly induced neuronal apoptosis characterized by cleaved caspase-3 expression, Hoechst 33342 staining, and intercellular Ca 2+ influx, whereas the presence of astrocytes significantly protected neurons from these effects. IFN-γ in the inflammation media also promoted astrocyte secretion of IL-6, essential for protection. The supernatants of rat peripheral blood mononuclear cells stimulated by lymphocyte mitogen lipopolysaccharide or concanavalin A were used as inflammation media to verify the results. The in vivo model involved a peripheral challenge with lipopolysaccharide, with or without recombinant IFN-γ, in C57BL/6 mice. This confirmed the in vitro results: anti-IFN-γ antibodies exacerbated the acute course of neuroinflammation and led to neurocyte apoptosis in vivo . The pro-inflammatory cytokine IFN-γ provided neuroprotection during acute neuroinflammation via induction of astrocyte-secreted IL-6. The findings provide novel insights into the mechanisms of neuroprotection by IFN-γ during acute neuroinflammation, and may impact therapies for inflammation-related central nervous system injury and disease.

// Yu-Lin Xu 1,* , Hai-Lin Tang 1,* , Hao-Ran Peng 1 , Ping Zhao 1 , Zhong-Tian Qi 1 and Wen Wang 1 1 Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University, Shanghai, China * These authors have contributed equally to this study Correspondence to: Zhong-Tian Qi, email: // Wen Wang, email: // Keywords : influenza A virus; H7N9 virus; RIP3; necroptosis; proinflammatory cytokines; Immunology and Microbiology Section; Immune response; Immunity Received : December 08, 2016 Accepted : February 23, 2017 Published : March 08, 2017 Abstract Influenza H7N9 virus infection causes an acute, highly contagious respiratory illness that triggers cell death of infected cells and airway epithelial destruction. RIP3 is a key regulator of cell death responses to a growing number of viral and microbial agents. This study aimed to investigate the role of RIP3 in inflammation of influenza H7N9 virus infection. Here, RIP3 knock out (RIP3-/-) mice and littermate wild type mice were infected intranasally with influenza H7N9 virus (A/Fujian/S03/2015) to determine the contribution of RIP3 to the inflammatory response of influenza H7N9 virus infection. It was found that RIP3-/- mice infected with H7N9 virus showed higher survival and less weight loss, compared with wild type littermate mice. In addition, RIP3-/- mice had fewer regions of edema, infiltration with inflammatory cells, and alveolar collapses, and the secretions of IL-1β, IL-6, RANTES and MIP-1 in BALF were significantly decreased on days 3 and 7 p.i. when compared with WT mice. Moreover, caspase 1/IL1β signaling was found to be involved in RIP3 associated inflammation of influenza H7N9 virus, but not RIP3/MLKL dependent necrosis. In the conclusion, our results indicated that RIP3 deficiency can protect mice from the infection of influenza H7N9 virus by downregulating caspase 1/IL1β signaling, which provided evidence of the RIP3 involved necroptosis independent manner.

// Yang Li 1,* , Jin Wang 1,* , Jie Yin 1 , Xinhua Liu 2 , Minghang Yu 1 , Ting Li 1 , Han Yan 1 and Xi Wang 1 1 Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Center of Basic Medical Sciences, Tianjin Key Laboratory of Medical Epigenetics, Tianjin Medical University, Tianjin, China 2 Department of Biochemistry and Molecular Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Tianjin Key Laboratory of Medical Epigenetics, School of Basic Medical Sciences, Tianjin Medical University, Tianjin, China * Yang Li and Jin Wang have contributed equally to this work Correspondence to: Xi Wang, email: // Keywords : NK cell, activation, histone modification, poised state, small molecule inhibitor, Immunology and Microbiology Section, Immune response, Immunity Received : December 08, 2016 Accepted : March 16, 2017 Published : March 29, 2017 Abstract Studies of Natural Killer (NK) cell cytotoxicity have mainly focused on the balance of activating and inhibitory receptors, signaling transduction, calcium influx, formation of immune synapse, and cytolytic degranulation. However, little is known about the chromatin state of NK cells and the impact of its changes during target recognition. In this study, we investigate the contribution of chromatin state dynamics during NK cell activation by comprehensively analyzing a set of microarray data and two sets of Chromatin Immunoprecipitation-Sequencing (ChIP-seq) data. We find that the expression of several histone demethylases and methyltransferases was influenced upon stimulation. Furthermore, we notice that a series of genes, including PI3KCA, NFATC1and TNFSF9, which play important roles during NK cell activation, were at ‘poised’ state prior to activation, and that modifications of H3K4me3 and H3K27me3 on these promotors were sensitive to stimulation with Phorbol Myristate Acetate (PMA) and Ionomycin (Iono) in the NK92MI cell line. Finally, we demonstrate that a series of small molecule inhibitors, which are specific to H3K4 and H3K27 modification, enhance degranulation or the expression levels of IFN-γ and TNF-α. Our results suggest that the histone modification state has a profound impact on NK cell activation, and provide novel insights into the regulation of NK cellular cytotoxicity and immunoregulatory function by chromatin state dynamics.

Degradation of lysosomal lipids requires lysosomal acid lipase (LAL), the only intracellular lipase known to be active at acidic pH. We found LAL to be expressed in murine immune cells with highest mRNA expression in macrophages and neutrophils. Furthermore, we observed that loss of LAL in mice caused lipid accumulation in white blood cells in the peripheral circulation, which increased in response to an acute inflammatory stimulus. Lal-deficient (-/-) macrophages accumulate neutral lipids, mainly cholesteryl esters, within lysosomes. The cholesteryl ester fraction is particularly enriched in the PUFAs 18:2 and 20:4, important precursor molecules for lipid mediator synthesis. To investigate whether loss of LAL activity affects the generation of lipid mediators and to eliminate potential systemic effects from other cells and tissues involved in the pronounced phenotype of Lal-/- mice, we treated macrophages from Wt mice with the LAL-specific inhibitor LAListat-2. Acute inhibition of LAL resulted in reduced release of 18:2- and 20:4-derived mediators from macrophages, indicating that lipid hydrolysis by LAL is an important source for lipid mediator synthesis in macrophages. We conclude that lysosomes should be considered as organelles that provide precursor molecules for lipid mediators such as eicosanoids.

// Hai Chen 1 , Xiaodong Qi 1 , Xue Bai 1 , Ping Qiu 1 , Bin Chen 1 1 Department of Galactophore, The General Hospital of Beijing Military Command, Beijing 100000, China Correspondence to: Hai Chen, email: yuijicshygu@163.com Keywords: CD152, CTLA-4, polymorphisms, breast cancer, immune response Received: July 02, 2016 Accepted: October 13, 2016 Published: March 01, 2017 ABSTRACT Background: The polymorphisms in cluster of differentiation 152 ( CD152 ) gene have been reported to be associated with breast cancer (BC), but relevant findings were far from conclusive. Therefore, we carried out this meta-analysis to combine those results for a clearer perspective on this issue. Results: In our meta-analysis, a total of 8 eligible publications of 19 case-control studies were selected, which totally contained 7,442 BC cases and 7,376 normal controls. Among the five polymorphisms of CD152 gene, +49 G/A, −1661 A/G and −318 C/T significantly increased the risk of BC under corresponding genetic comparisons; while CT60 G/A polymorphism was negatively related to the cancer susceptibility. In addition, −1772 T/C polymorphism of CD152 gene was not associated with the development of BC. Materials and methods: Online databases and other sources were searched for published studies on the relationship between BC susceptibility and CD152 polymorphisms (+49 G/A, −1661 A/G, −1722 T/C, −318 C/T and CT60 G/A). The strength of association was evaluated with pooled odds ratios (ORs) and their corresponding 95% confidence intervals (95% CIs). Heterogeneity evaluation was conducted via Q test. Sensitivity analysis was used to detect the stability of our results. Begg’s funnel plot and Egger’s test were applied to investigate publication bias among selected studies. Conclusions: The polymorphisms +49 G/A, −1661 A/G and −318 C/T may elevate the susceptibility to BC, but the polymorphism CT60 G/A may offer protection against the cancer.

// Si Chen 1,2,* , Bingni Chen 1,2,3,* , Zhongyang Wen 1,2,3,* , Zhong Huang 1,2,3 and Liang Ye 1,2,3 1 Institute of Biological Therapy, Shenzhen University, Shenzhen, China 2 Department of Pathogen Biology and Immunology, Shenzhen University School of Medicine, Shenzhen, China 3 Shenzhen City Shenzhen University Immunodiagnostic Technology Platform, Shenzhen, China * These authors have contributed equally to this work Correspondence to: Liang Ye, email: // Zhong Huang, email: // Keywords : IL-33, IL-33 receptor, IL-10, macrophage, rheumatoid arthritis, Immunology and Microbiology Section, Immune response, Immunity Received : August 11, 2016 Accepted : March 01, 2017 Published : March 16, 2017 Abstract IL-10 is an immunosuppressive cytokine produced and sensed by many immune cells and exerts a protective role in autoimmune diseases. However, the underlying mechanism by which IL-10 contributes to prevent the arthritic inflammation in macrophages is poorly understood. Herein we report on a novel anti-arthritic property of IL-10 through the inhibition of IL-33 signaling by macrophages during collagen-induced arthritis (CIA) development. We show that IL-33 expression rather than its receptor (ST2) is positively correlated with IL-10 level in active RA. IL-10 deficiency in mice leads to significant upregulation of IL-33 expression and aggravates the progression of CIA, while exogenous IL-10 treatment effectively diminishes IL-33 production in IL-10 knockout (IL-10 -/- ) CIA mice. We demonstrate further that the inhibitory effect of IL-10 in suppressing IL-33 production requires STAT3 activation in macrophages. Furthermore, IL-33 stimulated proinflammatory genes are notably increased in IL-10 -/- CIA mice, whereas macrophages treated with recombinant IL-10 exhibit decreased IL-33 amplified inflammation and inhibited IL-33 activated NF-κB signaling pathway. Our findings indicate that IL-10 act as a negative regulator of IL-33/ST2 signaling pathways in vivo , suggesting a potential therapeutic role of IL-10 in autoimmune diseases.

// Daisuke Muraoka 1,2 , Naohiro Seo 1 , Tae Hayashi 1 , Chisaki Hyuga-Amaike 1 , Kana Okamori 1 , Isao Tawara 3 , Naozumi Harada 1 and Hiroshi Shiku 1 1 Department of Immuno-Gene Therapy, Mie University Graduate School of Medicine, Mie, Japan 2 Center for Drug Discovery, Graduate School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan 3 Department of Hematology and Oncology, Mie University Graduate School of Medicine, Mie, Japan Correspondence to: Daisuke Muraoka, email: // Hiroshi Shiku, email: // Keywords : memory T cell, CTL function, STAP2, STAT signaling, SOCS3, Immunology and Microbiology Section, Immune response, Immunity Received : April 18, 2016 Accepted : February 01, 2017 Published : February 16, 2017 Abstract Long-surviving memory CD8 + T cells generated by stimulation with appropriate tumor-associated antigens are the most aggressive and persistent tumoricidal effectors. In this event of memory CD8 + T cell development, the signal transducer and activator of transcription (STAT) proteins function as the crucial intracellular signaling molecules, but the regulatory mechanism of STATs in CD8 + T cells is not fully understood. In this study, we report for the first time, by using murine vaccination models, that signal-transducing adaptor protein-2 (STAP2) maintains the cytotoxicity of long-lived memory CD8 + T cells by controlling a STAT3/suppressor of cytokine signaling 3 (SOCS3) cascade. Following T cell activation, STAP2 expression was transiently reduced but was subsequently recovered and augmented. Analysis using small-interfering RNA (siRNA) demonstrated that restored STAP2 expression was associated with the activation of STAT3/SOCS3 signals and maintenance of cytotoxic T lymphocytes (CTLs) secondary responses by preventing their differentiation into terminal effector cells. Notably, this STAP2-dependent memory differentiation was observed in the spleen, but not in the lymph nodes (LNs). These findings indicate an essential role for STAP2 in the generation of a high-quality memory CD8 + CTLs periphery, and suggest the therapeutic potential of STAP2 in cancer patients.

// Garima Garg 1 , Eirini Nikolouli 1 , Matthias Hardtke-Wolenski 2 , Aras Toker 1 , Naganari Ohkura 3,4 , Michael Beckstette 1 , Takahisa Miyao 5 , Robert Geffers 6 , Stefan Floess 1 , Norbert Gerdes 7,8 , Esther Lutgens 7,9 , Anke Osterloh 10 , Shohei Hori 5 , Shimon Sakaguchi 3,4 , Elmar Jaeckel 2 and Jochen Huehn 1 1 Department of Experimental Immunology, Helmholtz Centre for Infection Research, Braunschweig, Germany 2 Department of Gastroenterology, Hepatology, Endocrinology, Hannover Medical School, Hannover, Germany 3 Department of Experimental Immunology, World Premier International Immunology Frontier Research Center, Osaka University, Suita, Japan 4 Department of Experimental Pathology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan 5 Laboratory for Immune Homeostasis, RIKEN Center for Integrative Medical Sciences, Yokohama City, Kanagawa, Japan 6 Genome Analytics, Helmholtz Centre for Infection Research, Braunschweig, Germany 7 Institute for Cardiovascular Prevention, Ludwig-Maximilians-University, Munich, Germany 8 Division of Cardiology, Pulmonology, and Vascular Medicine Medical Faculty, University Hospital Dusseldorf, Dusseldorf, Germany 9 Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, AZ, Amsterdam, The Netherlands 10 Department of Immunology, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany Correspondence to: Jochen Huehn, email: // Keywords : regulatory T cells, thymic APCs, epigenetic modification, alloantigen-specificity, Immunology and Microbiology Section, Immune response, Immunity Received : October 15, 2016 Accepted : March 03, 2017 Published : March 15, 2017 Abstract Regulatory T cells (Tregs) are potential immunotherapeutic candidates to induce transplantation tolerance. However, stability of Tregs still remains contentious and may potentially restrict their clinical use. Recent work suggested that epigenetic imprinting of Foxp3 and other Treg-specific signature genes is crucial for stabilization of immunosuppressive properties of Foxp3 + Tregs, and that these events are initiated already during early stages of thymic Treg development. However, the mechanisms governing this process remain largely unknown. Here we demonstrate that thymic antigen-presenting cells (APCs), including thymic dendritic cells (t-DCs) and medullary thymic epithelial cells (mTECs), can induce a more pronounced demethylation of Foxp3 and other Treg-specific epigenetic signature genes in developing Tregs when compared to splenic DCs (sp-DCs). Transcriptomic profiling of APCs revealed differential expression of secreted factors and costimulatory molecules, however neither addition of conditioned media nor interference with costimulatory signals affected Foxp3 induction by thymic APCs in vitro . Importantly, when tested in vivo both mTEC- and t-DC-generated alloantigen-specific Tregs displayed significantly higher efficacy in prolonging skin allograft acceptance when compared to Tregs generated by sp-DCs. Our results draw attention to unique properties of thymic APCs in initiating commitment towards stable and functional Tregs, a finding that could be highly beneficial in clinical immunotherapy.

// Yuan Liu 1,* , Xining Liao 2,* , Ying Wang 3,* , Shiju Chen 1 , Yuechi Sun 1 , Qingyan Lin 1 and Guixiu Shi 1 1 Department of Rheumatology and Clinical Immunology, The First Affiliated Hospital of Xiamen University, Xiamen, China 2 Medical College, Xiamen University, Xiamen, Fujian, China 3 Department of Endocrinology, The ChengGong Hospital Affiliated to Xiamen University, Xiamen, Fujian, China * Yuan Liu, Xining Liao and Ying Wang contribute equally to this study Correspondence to: Guixiu Shi, email: // Keywords : primary Sjogren’s syndrome; anti-MDM2 autoantibody; biomarker;Immunology and Microbiology Section; Immune response; Immunity Received : September 27, 2016 Accepted : January 17, 2017 Published : January 28, 2017 Abstract Introduction: Primary Sjogren’s Syndrome (pSS) is one of the autoimmune diseases characterized by polyclonal autoantibody production. The human homologue of the mouse double minute 2 (MDM2) is an important negative regulator of p53. Our previous study indicated that autoantibody to MDM2 can be detected in systemic lupus erythematosus patients. The purpose of this study is to study anti-MDM2 autoantibody in pSS patients. Methods: Anti-MDM2 autoantibody in sera from 100 pSS patients and 74 normal controls was investigated by ELISA. Positive samples were further confirmed by western blotting. Expression of MDM2 in labial gland tissue from pSS patients and normal controls was checked by immunohistochemistry. The difference in clinical characteristics and laboratory findings between anti-MDM2 positive and anti-MDM2 negative pSS patients was analyzed. Results: The presence of anti-MDM2 autoantibody in pSS patients was 21.0%, significantly higher than normal controls (5.40%). MDM2 was overexpressed in labial gland from pSS patients. pSS patients with positive anti-MDM2 were characterized by longer disease duration and more lymphocytes focal gathering in labial gland. Prevalence of anemia, thrombocytopenia and anti-SSB was significantly higher in pSS patients with anti-MDM2 autoantibody. Titer of anit-MDM2 was negatively associated with hemoglobin level, platelet count, complement 3 level and complement 4 level, positively associated with European Sjogren’s syndrome disease activity index (ESSDAI) and level of IgG. Conclusions: Anti-MDM2 autoantibody may be used as a potential serological biomarker in pSS disease activity evaluation. Study on the role of anti-MDM2 or MDM2 in pSS may help us know the pathogenesis mechanism of pSS better.

Cancer gene therapy has attracted increasing attention for its advantages over conventional therapy in specific killing of tumor cells. Here, we attempt to prove a novel therapeutic approach that targets tumors by harnessing the blood antigen immune response system, which is inherently present in patients with breast cancers. Breast cancer MDA-MB-231 cells expressed blood group H antigen precursor. After ectopic expression of blood group A glycosyltransferase, we found that the H precursor was converted into the group A antigen, appearing on the surface of tumor cells. Incubation with group B plasma from breast cancer patients activated the antigen-antibody-complement cascade and triggered tumor cell killing. Interestingly, expression of blood A antigen also reduced tumorigenesis in breast cancer cells by inhibiting cell proliferation, migration, and tumor sphere formation. Cell cycle analysis revealed that cancer cells were paused at S phase due to the activation of cell cycle regulatory genes. Furthermore, pro-apoptotic genes were unregulated by the A antigen, including BAX, P21, and P53, while the anti-apoptotic BCL2 was down regulated. Importantly, we showed that extracellular HMGB1 and ATP, two critical components of the immunogenic cell death pathway, were significantly increased in the blood A antigen-expressing tumor cells. Collectively, these data suggest that blood antigen therapy induces specific cancer cell killing by activating the apoptosis and immunogenic cell death pathways. Further translational studies are thereby warranted to apply this approach in cancer immuno-gene therapy.

// Fengping Liu 1,4,* , Zongxin Ling 2,3,* , Yonghong Xiao 2,3,* , Longxian Lv 2,3 , Qing Yang 3 , Baohong Wang 2,3 , Haifeng Lu 2,3 , Li Zheng 1 , Ping Jiang 1 , Wei Wang 1,** and Lanjuan Li 2,3,** 1 Department of Urology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 2 State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, China 3 Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, China 4 Yancheng Medical College, Yancheng, Jiangsu, China * These authors have contributed equally to this work ** These authors also have contributed equally to this work Correspondence to: Wei Wang, email: // Lanjuan Li, email: // Keywords : Akkermansia muciniphila; Escherichia coli; lactobacillus; type 2 diabetes mellitus; urinary microbiota; Immunology and Microbiology Section, Immune response, Immunity Received : November 02, 2016 Accepted : December 13, 2016 Published : December 19, 2016 Abstract Type 2 diabetes mellitus (T2DM) may be associated with altered urinary microbiota in female patients. We investigated alterations of urinary microbiota in Chinese female T2DM patients, and explored the associations between urinary microbiota and a patient’s fasting blood glucose (FBG), urine glucose (UGLU), age, menstrual status, and body mass index (BMI). Midstream urine was collected from 70 female T2DM patients and 70 healthy females. Microbial diversity and composition were analyzed using the Illumina MiSeq sequencing platform by targeting the hypervariable V3-V4 regions of the 16S rRNA gene. We found that bacterial diversity was decreased in T2DM patients. Increased Actinobacteria phylum was positively correlated with FBG, UGLU, and BMI; Lactobacillus abundance decreased with age and menopause; and increased Lactobacillus correlated positively with FBG and UGLU. Decreased Akkermansia muciniphila was associated with FBG and UGLU. Escherichia coli abundance did not differ between the two cohorts. Carbohydrate and amino acid metabolism was reduced in T2DM patients, which were associated with bacterial richness indices such as Chao1 and ACE. Detailed microbiota analysis of well-characterized T2DM patients and healthy controls indicate that Chinese T2DM female patients exhibit dysbiosis of urinary microbiota.

// Juan Fu 1 , Rupashree Sen 1 , David L. Masica 2 , Rachel Karchin 2,3 , Drew Pardoll 3 , Vonn Walter 4 , D. Neil Hayes 5 , Christine H. Chung 6 and Young J. Kim 3,7 1 Department of Otolaryngology - Head & Neck Surgery, SKCCC, Johns Hopkins Hospital, Baltimore, MD, USA 2 Department of Biomedical Engineering and The Institute for Computational Medicine, SKCCC, Johns Hopkins Hospital, Baltimore, MD, USA 3 Bloomberg-Kimmel Institute for Cancer Immunotherapy, SKCCC, Johns Hopkins Hospital, Baltimore, MD, USA 4 Department of Biochemistry and Molecular Biology, Penn State Milton S. Hershey Medical Center, Hershey, PA, USA 5 UNC Chapel Hill School of Medicine, Lineberger Comprehensive Cancer Center, Chapel Hill, NC, USA 6 Department of Head & Neck - Endocrine Oncology, Moffitt Cancer Center, Tampa, FL, USA 7 Department of Otolaryngology - Head & Neck Surgery, VICC, Vanderbilt University Medical Center, Nashville, TN, USA Correspondence to: Young J. Kim, email: // Keywords : tumor microenvironment, autologous reconstitution, STAT3, humanized mice, head neck carcinoma, Immunology and Microbiology Section, Immune response, Immunity Received : August 19, 2016 Accepted : December 13, 2016 Published : December 19, 2016 Abstract Correlative studies from checkpoint inhibitor trials have indicated that better understanding of human leukocytic trafficking into the human tumor microenvironment can expedite the translation of future immune-oncologic agents. In order to directly characterize signaling pathways that can regulate human leukocytic trafficking into the tumor, we have developed a completely autologous xenotransplantation method to reconstitute the human tumor immune microenvironment in vivo. We were able to genetically mark the engrafted CD34+ bone marrow cells as well as the tumor cells, and follow the endogenous leukocytic infiltration into the autologous tumor. To investigate human tumor intrinsic factors that can potentially regulate the immune cells in our system, we silenced STAT3 signaling in the tumor compartment. As expected, STAT3 signaling suppression in the tumor compartment in these autologously reconstituted humanized mice showed increased tumor infiltrating lymphocytes and reduction of arginase-1 in the stroma, which were associated with slower tumor growth rate. We also used this novel system to characterize human myeloid suppressor cells as well as to screen novel agents that can alter endogenous leukocytic infiltration into the tumor. Taken together, we present a valuable method to study individualized human tumor microenvironments in vivo without confounding allogeneic responses.

// David J. Paulucci 1 , John P. Sfakianos 1 , Anders J. Skanderup 2 , Kathleen Kan 1 , Che-Kai Tsao 3 , Matthew D. Galsky 3 , A. Ari Hakimi 4 , Ketan K. Badani 1 1 Department of Urology, Icahn School of Medicine at Mount Sinai Hospital, New York, NY, USA 2 Computational Biology Program, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, New York, NY, USA 3 Department of Medicine, Division of Hematology/Oncology, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA 4 Department of Surgery—Urology Service, Memorial Sloan Kettering Cancer Center, New York, NY, USA Correspondence to: Ketan K. Badani, email: Ketan.Badani@mountsinai.org Keywords: papillary renal cell carcinoma, racial disparities, immune system signaling, targeted therapy, immune response Received: May 03, 2016 Accepted: November 22, 2016 Published: December 23, 2016 ABSTRACT Significant disparities in survival, incidence and possibly response to current therapies exist between black and white patients with renal cell carcinoma (RCC). Recent genomic evidence to account for these disparities has been reported for clear cell RCC. However, racial disparities at the genomic level for papillary RCC (pRCC) which is a genetically distinct and less responsive histologic subtype of RCC have not been reported. Using The Cancer Genome Atlas (TCGA) data, the present study assessed gene-level expression, somatic mutation and pathway differences between 58 black and 58 white patients with pRCC propensity matched on age, gender and pathologic T stage. Distinct tumor biology with differential expression patterns were observed in black vs. white patients with pRCC. Specifically, significance analysis of microarrays was applied to TCGA gene expression data and identified 163 genes and 120 genes overexpressed in black and white patients, respectively (FDR q

// Yingfeng Wei 1 , Chong Lu 1 , Jianing Chen 1 , Guangying Cui 1 , Lin Wang 1 , Tianming Yu 2 , Yue Yang 3 , Wei Wu 1 , Yulong Ding 1 , Lanjuan Li 1 , Toshimitsu Uede 4 , Zhi Chen 1 and Hongyan Diao 1 1 State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 2 Department of Orthopaedics, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 3 Department of Biochemistry, University of Washington, Seattle, Washington, USA 4 Molecular Immunology, Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan Correspondence to: Hongyan Diao, email: // Zhi Chen, email: // Keywords : high salt diet, Th17 cells, regulatory T cells, inflammatory bowel diseases, intestinal immunity, Immunology and Microbiology Section, Immune response, Immunity Received : August 01, 2016 Accepted : November 24, 2016 Published : December 01, 2016 Abstract This study focuses on characterizing the effect of a high salt diet (HSD) on intestinal immunity and the risk of inflammatory bowel diseases (IBD). We found that mice on a HSD had an increased frequency of IL-17A producing cells in the intestinal lamina propria (LP) compared to mice on a normal diet (ND). Furthermore, most intestinal IL-17A producing cells were CD4 + TCRβ + cells. A HSD increased the LP T helper 17 (Th17) responses in both the small and large intestines but did not increase the Th17 response of other gut-associated lymphoid organ. Although, HSD did not change the percentage of regulatory T (Treg) cells, HSD significantly inhibit secretion of IL-10 and the suppressive function of Treg cells. Moreover, we found that HSD exacerbates trinitrobenzenesulfonic acid (TNBS) induced colitis, and Th17 response was significantly increased in the colonic LP of HSD TNBS-treated mice compared with the ND TNBS-treated mice. This study demonstrates that HSD stimulates the intestinal Th17 response but inhibits the function of Treg cells. Moreover, HSD exacerbates TNBS induced mice colitis, suggesting that HSD disrupts the intestinal immunity and increases the risk of IBD.

// Weiping Hu 1,* , Guodong Niu 1,* , Hongbo Li 1 , Hanyuan Gao 1 , Rudian Kang 1 , Xiaoqing Chen 1 and Ling Lin 1 1 Rheumatism Department, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian Province, China * These authors have contributed equally to this study Correspondence to: Xiaoqing Chen, email: // Ling Lin, email: // Keywords : IFIT1, podocytes, animal study, renal pathological changes, lupus nephritis, Immunology and Microbiology Section, Immune response, Immunity Received : July 28, 2016 Accepted : October 28, 2016 Published : November 03, 2016 Abstract Renal damage is the major cause of SLE associated mortality, and IFIT1 expression was elevated in SLE cases in accordance of previous studies. Therefore, we conducted an animal study to identify the role of IFIT1 expression in renal pathological changes.18 female MRL/lpr mice and same number of female BALB/c mice were enrolled in present study. Quantitative analysis of urine protein, Complement C3 and C4, and anti-ds DNA antibody were conducted. HE and PAS staining and TEM analysis were employed to observe the pathological changes in renal tissue. Significant elevation on urine protein and anti-dsDNA and reduction on Complement C3 and C4 were observed in MRL/lpr mice when comparing the controls in same age. Staining and TEM analysis observed several pathological changes in glomerulus among MRL/lpr mice, including cellular enlargement, basement membrane thickening, and increased cellularcasts. The linear regression analysis found the optical density of IFIT1 was inversely associated with F-actin, Nephrin, and Podocin, but not Synatopodin. In summary, IFIT1 expression is associated with podocytes damage, and capable of suppressing some proteins essential to glomerular filtration.

Viruses with segmented RNA genomes must package the correct number of segments for synthesis of infectious virus particles. Recent studies suggest that the members of the Reoviridae family with segmented double-stranded RNA genomes achieve this challenging task by forming RNA networks of segments prior to their recruitment into the assembling capsid albeit direct evidence is still lacking. Here, we investigated the capability of virus recovery by preformed complexes of ten RNA segments of Epizootic Haemorrhagic Disease Virus (EHDV), a Reoviridae member, by transcribing exact T7 cDNA copies of genomic RNA segments in a single in vitro reaction followed by transfection of mammalian cells. The data obtained was further confirmed by RNA complexes generated from Bluetongue virus, another family member. Formation of RNA complexes was demonstrated by sucrose gradient ultracentrifugation, and RNA-RNA interactions inherent to the formation of the RNA complexes were demonstrated by electrophoretic mobility shift assay. Further, we showed that disruption of RNA complex formation inhibits virus recovery, confirming that recruitment of complete RNA networks is essential for packaging and consequently, virus recovery. This efficient reverse genetics system will allow further understanding of evolutionary relationships of Reoviridae members and may also contribute to development of antiviral molecules.

// Hyunjung Baek 1 , Minsook Ye 1 , Geun-Hyung Kang 1 , Chanju Lee 1 , Gihyun Lee 1 , Da Bin Choi 1 , Jaehoon Jung 1 , Hyunseong Kim 2 , Seonhwa Lee 3 , Jin Su Kim 3 , Hyun-ju Lee 4 , Insop Shim 4 , Jun-Ho Lee 5 and Hyunsu Bae 1 1 Department of Physiology, College of Korean Medicine, Kyung Hee University, Seoul, Republic of Korea 2 K-herb Research Center, Korea Institute of Oriental Medicine, Yuseongdae-ro, Yuseong-gu, Daejeon, Republic of Korea 3 Division of RI Research, Korea Institute of Radiological & Medical Sciences University of Science & Technology, Gongneug-dong, Nowon-ku, Seoul, Republic of Korea 4 Acupuncture and Meridian Science Research Center, College of Korean Medical Science Graduate School, Kyung Hee University, Seoul, Republic of Korea 5 Department of Biotechnology, Chonnam National University, Gwangju, Republic of Korea Correspondence to: Hyunsu Bae, email: // Keywords : Alzheimer’s disease, regulatory T cells (Treg), amyloid-beta (Aβ) pathology, Immunology and Microbiology Section, Immune response, Immunity Received : July 23, 2016 Accepted : September 29, 2016 Published : October 04, 2016 Abstract Alzheimer’s disease patients display neuropathological lesions, including the accumulation of amyloid-beta (Aβ) peptide and neurofibrillary tangles. Although the mechanisms causing the neurodegenerative process are largely unknown, increasing evidence highlights a critical role of immunity in the pathogenesis of Alzheimer’s disease. In the present study, we investigated the role of regulatory T cells (Tregs) on Alzheimer’s disease progression. First, we explored the effect of Tregs (CD4 + CD25 + T cells) and Teffs (CD4 + CD25 - T cells) in an adoptive transfer model. Systemic transplantation of purified Tregs into 3xTg-AD mice improved cognitive function and reduced deposition of Aβ plaques. In contrast, adoptive transfer of Teffs diminished behavioral function and cytokine production. Next, we transiently depleted Treg population using an anti-CD25 antibody (PC61). Depletion of Tregs for four months resulted in a marked aggravation of the spatial learning deficits of six-month-old 3xTg-AD mice. Additionally, it resulted in decreasing glucose metabolism, as assessed by positron emission tomography (PET) with 18 F-2 fluoro-2-deoxy-D-glucose ([F- 18 ] FDG) neuroimaging. Importantly, the deposition of Aβ plaques and microglia/macrophage was increased in the hippocampal CA1 and CA3 regions of the Treg depleted 3xTg-AD compared to the vehicle-treated 3xTg-AD group. Our finding suggested that systemic Treg administration ameliorates disease progression and could be an effective Alzheimer’s disease treatment.

// Ruya Zhang 1,* , Fang Ni 1,2,* , Binqing Fu 1 , Yang Wu 1 , Rui Sun 1 , Zhigang Tian 1 and Haiming Wei 1 1 Institute of Immunology and the CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Life Science and Medical Center, University of Science and Technology of China, Hefei National Laboratory for Physical Sciences at Microscale, University of Science and Technology of China, Hefei, China 2 Department of Pathophysiology, Anhui Medical University, Hefei, Anhui, China * These authors have contributed equally to this work Correspondence to: Haiming Wei, email: // Zhigang Tian, email: // Keywords : long noncoding RNAs, natural killer cells, CD56, primary lymphocytes, lnc-CD56, Immunology and Microbiology Section, Immune response, Immunity Received : May 06, 2016 Accepted : September 28, 2016 Published : October 04, 2016 Abstract Natural killer (NK) cells are innate immune lymphocytes that play critical roles in host defense against viral infection and surveillance against malignant transformation. Long noncoding RNAs (lncRNAs) are important immune system regulators. Here, we analyzed human primary lymphocyte lncRNA expression profiles to identify NK-lncRNA signatures. We detected numerous novel NK-specific lncRNAs with potential roles in regulating human NK cell differentiation and function. Expression of lnc-CD56, an NK-specific lncRNA, was positively correlated with that of CD56, a classical human NK cell surface marker. We showed that lnc-CD56 may function as a positive regulator of CD56 in primary human NK cells and differentiated NK cells from human CD34 + hematopoietic progenitor cells. Our data provide an annotated human NK cell lncRNA expression catalog and demonstrate a key role for lncRNAs in NK cell biology.

Guillain-Barré syndrome (GBS) is an autoimmune-mediated peripheral neuropathy of unknown cause. However, about a quarter of GBS patients have suffered a recent bacterial or viral infection, and axonal forms of the disease are especially common in these patients. Proteomics is a good methodological approach for the discovery of disease biomarkers. Until recently, most proteomics studies of GBS and other neurodegenerative diseases have focused on the analysis of the cerebrospinal fluid (CSF). However, serum represents an attractive alternative to CSF because it is easier to sample and has potential for biomarker discovery. The goal of this research was the identification of serum biomarkers associated with recovery from GBS. To address this objective, a quantitative proteomics approach was used to characterize differences in the serum proteome between a GBS patient and her healthy identical twin in order to lessen variations due to differences in genetic background, and with additional serum samples collected from unrelated GBS (N = 3) and Spinal Cord Injury (SCI) (N = 3) patients with similar medications. Proteomics results were then validated by ELISA using sera from additional GBS patients (N = 5) and healthy individuals (N = 3). All GBS and SCI patients were recovering from the acute phase of the disease. The results showed that Piccolo, a protein that is essential in the maintenance of active zone structure, constitutes a potential serological correlate of recovery from GBS. These results provided the first evidence for the Piccolo's putative role in GBS, suggesting a candidate target for developing a serological marker of disease recovery., This work was partially supported by EU FP7 ANTIGONE project number 278976. LMH was supported by a fellowship from the University of Castilla La Mancha (UCLM), Spain. MV was supported by the Research Plan of UCLM, Spain.

// Kelly C. G. Manfrere 1 , Marina P. Torrealba 1 , Denis R. Miyashiro 2 , Luanda M. S. Oliveira 1 , Gabriel C. de Carvalho 1 , Josenilson F. Lima 1 , Anna Claudia C. C. Branco 1 , Natalli Z. Pereira 1 , Juliana Pereira 3 , Jose A. Sanches, Jr 2,* and Maria N. Sato 1,* 1 Department of Dermatology, Laboratory of Medical Investigation, LIM 56, Tropical Medicine Institute of Sao Paulo, University of Sao Paulo Medical School, Brazil 2 Department of Dermatology, Cutaneous Lymphoma Clinic, Hospital das Clinicas, University of Sao Paulo, Medical School, Brazil 3 Department of Hematology, University of Sao Paulo Medical School, Brazil * These authors share the mentorship, critical revision and supervision of this study Correspondence to: Maria N. Sato, email: // Keywords : Se,zary syndrome, innate immunity, Toll-like receptor agonists, cytokines, type I interferon, Immunology and Microbiology Section, Immune response, Immunity Received : July 28, 2016 Accepted : October 14, 2016 Published : October 21, 2016 Abstract Sezary syndrome (SS) carries a poor prognosis, and infections represent the most frequent cause of death in SS patients. Toll-like receptors (TLRs) are a family of innate immune receptors that induce protective immune responses against infections. We sought to evaluate the ability of TLR agonists to induce inflammatory cytokine, Th2 cytokine, and type I interferon (IFN-I) production by peripheral blood mononuclear cells (PBMC) of untreated SS patients. We detected impaired IL-6, IL-10 and IL-13 secretion by PBMC induced by the agonists for TLR5, TLR3, TLR7 and TLR9 in SS patients, while it was partially recovered by TLR2/TLR4 and TLR7/8 agonists TNF secretion was restored following stimulation with TLR2/TLR4 agonists. IFN-γ was scarcely produced upon TLR activation in SS cells, albeit TLR 7/8 (CL097) enhanced their secretion at lower levels than the control group. TLR9 agonist efficiently induced IFN-I in SS patients, although this positive regulation was not observed for other cytokines, in direct contrast to the broad activity of CL097. Among the TLR agonists, TLR4 was able to induce pro-inflammatory, IL-10 and Th2 secretion, while TLR7-8 agonist induced the inflammatory cytokines, IFN-I and IFN-γ. These findings reveal a dysfunctional cytokine response upon both extracellular and intracellular TLR activation in SS patients, which was partially restored by TLRs agonists.

The immunoregulatory function of T regulatory cells (Tregs) is impaired in multiple sclerosis (MS). Recent studies have shown that umbilical cord-derived mesenchymal stem cells (UC-MSCs) exert regulatory effect on the functions of immune cells. Thus, we investigated whether UC-MSCs could improve the impaired function of Tregs from MS patients. Co-cultures of UC-MSCs with PBMCs of MS patients were performed for 3 days. Flow cytometry was used to determine the frequency of Tregs. A cell proliferation assay was used to evaluate the suppressive capacity of Tregs. ELISA was conducted for cytokine analysis in the co-cultures. Our results showed that UC-MSCs significantly increased the frequency of CD4+CD25+CD127low/- Tregs in resting CD4+ T cells (p

// Andrea Petretto 1,* , Grazia Carbotti 2,* , Elvira Inglese 1 , Chiara Lavarello 1 , Maria Pia Pistillo 3 , Valentina Rigo 2 , Michela Croce 2 , Luca Longo 2 , Stefania Martini 2 , Paola Vacca 2,4 , Silvano Ferrini 2,* and Marina Fabbi 2,* 1 Core Facilities-Proteomics Laboratory, Istituto Giannina Gaslini, Genoa, Italy 2 Department of Integrated Oncological Therapies, IRCCS AOU San Martino-IST Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy 3 Tumor Epigenetics Unit, IRCCS AOU San Martino-IST Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy 4 Department of Experimental Medicine (DIMES), University of Genoa, Genoa, Italy * These authors have equally contributed to this work Correspondence to: Silvano Ferrini, email: // Marina Fabbi, email: // Keywords : IL-27, IFN-γ, proteomic analysis, HLA class I, cancer cells, Immunology and Microbiology Section, Immune response, Immunity Received : July 10, 2016 Accepted : September 17, 2016 Published : September 24, 2016 Abstract IL-27, a member of the IL-12-family of cytokines, has shown anti-tumor activity in several pre-clinical models due to anti-proliferative, anti-angiogenic and immune-enhancing effects. On the other hand, IL-27 demonstrated immune regulatory activities and inhibition of auto-immunity in mouse models. Also, we reported that IL-27, similar to IFN-γ, induces the expression of IL-18BP, IDO and PD-L1 immune regulatory molecules in human cancer cells. Here, a proteomic analysis reveals that IL-27 and IFN-γ display a broad overlap of functions on human ovarian cancer cells. Indeed, among 990 proteins modulated by either cytokine treatment in SKOV3 cells, 814 showed a concordant modulation by both cytokines, while a smaller number (176) were differentially modulated. The most up-regulated proteins were common to both IFN-γ and IL-27. In addition, functional analysis of IL-27-regulated protein networks highlighted pathways of interferon signaling and regulation, antigen presentation, protection from natural killer cell-mediated cytotoxicity, regulation of protein polyubiquitination and proteasome, aminoacid catabolism and regulation of viral protein levels. Importantly, we found that IL-27 induced HLA class I molecule expression in human cancer cells of different histotypes, including tumor cells showing very low expression. IL-27 failed only in a cancer cell line bearing a homozygous deletion in the B2M gene. Altogether, these data point out to a broad set of activities shared by IL-27 and IFN-γ, which are dependent on the common activation of the STAT1 pathway. These data add further explanation to the anti-tumor activity of IL-27 and also to its dual role in immune regulation.

Astrocytes play a crucial role in neuroinflammation as part of the glia limitans, which regulates infiltration of the brain parenchyma by leukocytes. The signaling pathways and molecular events, which result from the interaction of activated T cells with astrocytes are poorly defined. Here we show that astrocytes promote the expression and enzymatic activity of CD39 and CD73 ectonucleotidases in recently activated CD4 cells by a contact dependent mechanism that is independent of T cell receptor interaction with class II major histocompatibility complex (MHC). Transforming growth factor-β (TGF-β) is robustly upregulated and sufficient to promote ectonucleotidases expression. T cell adhesion to astrocyte results in differentiation to an immunosuppressive phenotype defined by expression of the transcription factor Rorγt, which characterizes the CD4 T helper 17 subset. CD39 activity in T cells in turn inhibits spontaneous calcium oscillations in astrocytes that correlated with enhanced and reduced transcription of CCL2 chemokine and Sonic hedgehog (Shh), respectively. We hypothesize this TCR-independent interaction promote an immunosuppressive program in T cells to control possible brain injury by deregulated T cell activation during neuroinflammation. On the other hand, the increased secretion of CCL2 with concomitant reduction of Shh might promote leukocytes extravasation into the brain parenchyma.