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The synthetic oligonucleotide SD-101 is a potent and specific agonist for toll-like receptor 9. Intratumoral injection of SD-101 induces significant anti-tumor immunity in preclinical and clinical studies, especially when combined with PD-1 blockade. To build upon this strategy, we studied the enhancement of SD-101 activities by combination with low-dose cyclophosphamide, a well-characterized agent with potentially complementary activities. In multiple mouse tumor models, we demonstrate substantial anti-tumor activity of the combination, compared to each single agent. Combination therapy generated CD8+ T cell dependent immunity leading to rejection of both non-injected and injected tumors and long-term survival, even in very large tumors. Mechanistic studies encompassing global gene expression changes and characterization of immune cell infiltrates show the rapid, sequential induction of innate and adaptive responses and identify discrete contributions of SD-101 and cyclophosphamide. Importantly, these changes were prominent in tumors not injected directly with SD-101. Combination treatment resulted in creation of a permissive environment for a systemic anti-tumor immune response, including a reduction of intratumoral regulatory T cells (Tregs) and an increase in “M1” versus “M2” tumor-associated macrophage (TAM) phenotypes. Additionally, we observed increased immunogenic cell death as well as antigen processing in response to combination treatment.

Safety evaluation of a human vaccine is critical for vaccine development and for preventing an unexpected adverse reaction in humans. Nonetheless, to date, very few systems have been described for preclinical studies of human adverse reactions in vivo. Previously, we have identified biomarker genes expressed in the lungs for evaluation of influenza vaccine safety, and their usefulness in rodent models and for adjuvant-containing vaccines has already been reported. Here, our purpose was to develop a novel humanized mouse model retaining human innate-immunity-related cells to assess the safety of influenza vaccines using the previously identified biomarker genes. In the present study, we tested whether the two humanized models, a short-term and long-term reconstitution model of NOD/Shi-scid IL2rγnull mice, are suitable for biomarker gene-based safety evaluation. In the short-term model, human CD14+ cells, plasmacytoid dendritic cells, CD4+ and CD8+ T cells, and B cells were retained in the lungs. Among these cells, human CD14+ cells and plasmacytoid dendritic cells were not detected in the lungs of the long-term model. After the vaccination, the expression levels of human biomarker genes were elevated only in the short-term model when the toxicity reference vaccine was inoculated. This phenomenon was not observed in the long-term model. The levels of human cytokines and chemokines in the lungs increased in response to the toxicity reference vaccine in the short-term mouse model. According to these results, the short-term model provides a better platform for evaluating vaccine safety in terms of human peripheral blood mononuclear cell-mediated initial reactions in vivo.

// Jonathan Lalsiamthara 1 , Je Hyoung Kim 1 and John Hwa Lee 1 1 College of Veterinary Medicine, Chonbuk National University, Iksan Campus, Iksan 54596, Republic of Korea Correspondence to: John Hwa Lee, email: johnhlee@jbnu.ac.kr Keywords: Salmonella Typhimurium; live-attenuated bacterial vector; pre-existing immunity; heterologous antigen delivery Received: September 05, 2017 Accepted: April 05, 2018 Published: May 22, 2018 ABSTRACT Live Salmonella vaccine vectors offer a remarkable platform for delivering immunogens and therapeutic molecules by mimicking natural intracellular infections; however, pre-existing anti-vector immunity can impede effective deployment. Measures to alleviate pre-existing immunity include the use of heterologous vectors, development of highly attenuated strain enabling greater payload, removal of major immunoreactive components from the vector, and/or augmentation of delivered antigens via increased presentation in antigen presenting cells. Here we report a Salmonella Typhimurium (ST) vector-JOL1800 that embodies these requisite properties. JOL1800 is a highly attenuated, auxotrophic, and O-antigen deficient rough-mutant strain. Heterologous bacterial and viral antigens were expressed and delivered using JOL1800 in mice, irrespective of the inoculation route successful inductions of the mucosal and systemic humoral responses were observed. Compared to smooth LPS vector delivery, we observed an increased fraction of delivered-antigen presenting dendritic cells and a higher frequency of delivered-antigen displayed per macrophage. Upon post-priming with JOL1800 delivery, efficacy of the delivery was minimally affected as indicated by insignificant decrease in colonization, humoral and cellular responses. Our results show that the generated vector is capable of remote antigen delivery, manifests higher antigen presentation, is Differentiating Infected from Vaccinated Animals (DIVA) capable, evades normal pre-existing immunity, and can be deployed for effective delivery.

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

// Tatsuo Kanda 1, 2 , Shin Yasui 2 , Masato Nakamura 2 , Shingo Nakamoto 2 , Koji Takahashi 2 , Shuang Wu 2 , Reina Sasaki 2 , Yuki Haga 2 , Sadahisa Ogasawara 2 , Tomoko Saito 2 , Kazufumi Kobayashi 2 , Soichiro Kiyono 2 , Yoshihiko Ooka 2 , Eiichiro Suzuki 2 , Tetsuhiro Chiba 2 , Hitoshi Maruyama 2 , Fumio Imazeki 2 , Mitsuhiko Moriyama 1 and Naoya Kato 2 1 Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, Tokyo 173-8610, Japan 2 Department of Gastroenterology, Chiba University, Graduate School of Medicine, Chiba 260-8670, Japan Correspondence to: Tatsuo Kanda, email: kandat-cib@umin.ac.jp Keywords: hepatitis C virus; direct-acting antivirals; autoimmune hepatitis; primary biliary cholangitis; sustained virological response Received: January 16, 2018 Accepted: January 30, 2018 Published: February 03, 2018 ABSTRACT Background: Interferon-free treatment can achieve higher sustained virological response (SVR) rates, even in patients in whom hepatitis C virus (HCV) could not be eradicated in the interferon treatment era. Immune restoration in the liver is occasionally associated with HCV infection. We examined the safety and effects of interferon-free regimens on HCV patients with autoimmune liver diseases. Results: All 7 HCV patients with autoimmune hepatitis (AIH) completed treatment and achieved SVR. Three patients took prednisolone (PSL) at baseline, and 3 did not take PSL during interferon-free treatment. In one HCV patient with AIH and cirrhosis, PSL were not administered at baseline, but she needed to take 40 mg/day PSL at week 8 for liver dysfunction. She also complained back pain and was diagnosed with vasospastic angina by coronary angiography at week 11. However, she completed interferon-free treatment. All 5 HCV patients with primary biliary cholangitis (PBC) completed treatment and achieved SVR. Three of these HCV patients with PBC were treated with UDCA during interferon-free treatment. Conclusions: Interferon-free regimens could result in higher SVR rates in HCV patients with autoimmune liver diseases. As interferon-free treatment for HCV may have an effect on hepatic immunity and activity of the autoimmune liver diseases, careful attention should be paid to unexpected adverse events in their treatments. Methods: Total 12 patients with HCV and autoimmune liver diseases [7 AIH and PBC], who were treated with interferon-free regimens, were retrospectively analyzed.

In this study, experimental pathology, flow cytometry (FCM), quantitative real-time polymerase chain reaction (qRT-PCR), and western blot (WB) were used to evaluate the effects of sodium fluoride (NaF) on hepatocellular cell cycle progression in mice. A total of 240 ICR mice were divided equally into four groups; the experimental groups received 12, 24, or 48 mg/kg NaF intragastrically for 42 days, while the control group received distilled water. Doses of NaF above 12 mg/kg increased the percentage of cells in S phase (S-phase arrest), reduced percentages of cells in G0/G1 or G2/M phase, and activated the ATM-p53-p21 and ATR-Chk1-Cdc25A pathways. Activation of these pathways was characterized by up-regulation of ATM, p53, p21, ATR, and Chk1 mRNA and protein expression, and down-regulation of Cdc25A, cyclin E, cyclin A, CDK2, CDK4, and proliferating cell nuclear antigen (PCNA) mRNA and protein expression. These results indicate that NaF caused S-phase arrest by activating the ATM-p53-p21 and ATR-Chk1-Cdc25A pathways.

// Ming-Jie Wang 1,* , Xiao-Liang Xu 2,* , Guo-Liang Yao 3,* , Qiang Yu 4 , Chun-Fu Zhu 4 , Zhi-Jun Kong 4 , Hui Zhao 5 , Li-Ming Tang 4 and Xi-Hu Qin 4 1 Department of Orthopedics, Affiliated Hospital of Nanjing Medical University, Changzhou Second People’s Hospital, Changzhou, China 2 Liver Surgery of Jiangsu Province People’s Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China 3 Department of Thoracic Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China 4 Department of General Surgery, Affiliated Hospital of Nanjing Medical University, Changzhou Second People’s Hospital, Changzhou, China 5 Department of General Surgery, Wuxi Third People’s Hospital, Wuxi, China * These authors have contributed equally to this study Correspondence to: Zhi-Jun Kong, email: // Hui Zhao, email: // Xi-Hu Qin, email: // Keywords : inflammatory bowel disease; Crohn’s disease; ulcerative colitis; MYO9B; polymorphism; Immunology and Microbiology Section; Immune response; Immunity Received : March 25, 2016 Accepted : July 19, 2016 Published : August 19, 2016 Abstract Myosin IXB ( MYO9B ) gene polymorphisms have been extensively investigated in terms of their associations with inflammatory bowel disease (IBD), with contradictory results. The aim of this meta-analysis was to evaluate associations between MY09B gene polymorphisms and the risk of IBD, Crohn’s disease (CD) and ulcerative colitis (UC). Eligible studies from PubMed, Embase, and CNKI databases were identified. Pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. Ten studies published in eight papers reporting 8,975 cases and 9,482 controls were included in this meta-analysis. Five MY09B gene polymorphisms were evaluated: rs1545620, rs962917, rs1457092, rs2305764, and rs2305767. Our data suggested that the rs1545620 polymorphism was associated with a decreased risk of IBD. A similar result was found for rs2305767 and UC. The rs962917 single nucleotide polymorphism (SNP) increased the risk of IBD, CD and UC. Moreover, rs1457092 increased the risk of IBD and UC. Rs2305764 was also associated with an increased risk of IBD. Furthermore, stratification analyses indicated that rs1545620 decreased the risk of IBD, while rs962917 increased the risk of IBD, CD and UC in Caucasian populations. To sum up, our data indicate that these five SNPs in MY09B are significantly associated with the risk of IBD.

Tumor-associated carbohydrate antigens (TACAs) support cell survival that could be interrupted by anti-TACA antibodies. Among TACAs that mediate cell survival signals are the neolactoseries antigen Lewis Y (LeY) and the ganglioside GD2. To induce sustained immunity against both LeY and GD2, we developed a carbohydrate mimicking peptide (CMP) as a surrogate pan-immunogen that mimics both. This CMP, referred to as P10s, is the N-terminal half of a peptide vaccine named P10s-PADRE, the C-terminal half of which (PADRE) is a Pan-T-cell epitope. A Phase I dose-escalation trial of P10s-PADRE plus adjuvant MONTANIDE™ ISA 51 VG was conducted in subjects with metastatic breast cancer to test 300 and 500 μg/injection in two cohorts of 3 subjects each. Doses of the P10s-PADRE vaccine were administered to research participants subcutaneously on weeks 1, 2, 3, 7 and 19. Antibody responses to P10s, GD2, and LeY were measured by ELISA. The P10s-PADRE vaccine induced antibodies specifically reactive with P10s, LeY and GD2 in all 6 subjects. Serum antibodies displayed Caspase-3-dependent apoptotic functionality against LeY or GD2 expressing breast cancer cell lines. Immunization with the P10s-PADRE vaccine was well-tolerated and induced functional antibodies, and the data suggest potential clinical benefit.

// Yu-Jie Wang 1,2,* , Lin Li 1,4,* , Wei-Juan Song 3,* , Yan-Jun Zhou 1 , Meng-Da Cao 1 , Xiang-Rong Zuo 1,2 and Ji-Fu Wei 1 1 Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China 2 Department of Critical Care Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China 3 Department of Clinical Laboratory, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China 4 Department of Emergency Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China * These authors have contributed equally to this article Correspondence to: Xiang-Rong Zuo, email: // Ji-Fu Wei, email: // Keywords : Can f 6, dog allergy, epitopes, allergenicity, Chinese children, Immunology and Microbiology Section, Immune response, Immunity Received : July 07, 2017 Accepted : September 20, 2017 Published : October 13, 2017 Abstract Dog allergy is common worldwide. However, the allergenicity of dog allergy is still unclear in China as well as in special group, such as children. In this study, we chose Can f 6, a major dog allergen which belongs to the lipocalin to study its allergenicity in Chinese dog allergic children. Can f 6 gene was subcloned into pET-28a vector and transformed into E. coli BL21 (DE3) cells for expression. The recombinant Can f 6 was purified by nickel affinity chromatography, identified by SDS-PAGE, and tested for its allergenicity by Western blot with sera and basophil activation test. Secondary structures, B cell epitopes and homology modeling of Can f 6 were predicted by using a series of bioinformatical approaches. And the verification of B cell epitopes was detected by ELISA. The recombinant allergen showed an explicit band with the molecular weight of 20 kDa by SDS-PAGE. Sera from 56.3 % (18/32) of dog-allergic children patients reacted with Can f 6. The induction of the expression of CD63 and CCR3 of dog allergic children in passively sensitized basophils was up to approximately 5.0 times higher than healthy subjects. The secondary structure of Can f 6 contains 3 α-helices, 9 β-sheets and random coils. Five B cell epitopes of Can f 6 were predicted and were confirmed successfully by ELISA. The results showed Can f 6 is a major allergen in Chinese children, which provides a basis for further study of Can f 6 in diagnosis and treatment of symptoms in children in China. The structural information of Can f 6 will help to form a foundation for the future design of vaccines and therapies for Can f 6 related allergies.

In osteoarthritis (OA), activated synoviocytes and articular chondrocytes produce pro-inflammatory cytokines, such as IL-1β, that promote chondrocyte apoptosis and activate the NF-κB signaling pathway to induce catabolic factors. In this study, we examined the anti-inflammatory and anti-apoptotic effect of baicalein on IL-1β signaling and NF-κB-regulated gene products in rat chondrocytes. Rat chondrocytes were pretreated with 10 ng/ml IL-1β for 24 h and then co-treated with 10 ng/ml IL-1β and 50 μM baicalein for 0, 12, 24, 36 and 48h. The expression levels of poly(ADP-ribose) polymerase (PARP), Bcl-2, caspase-3, matrix metalloproteinase (MMP)-9, MMP-3, cyclooxygenase (COX)-2 and SOX-9 were detected by Western blot and quantitative reverse transcription-PCR (qPCR). The effects of baicalein on the translocation and phosphorylation of the NF-κB system were studied by Western blotting and immunofluorescence. Baicalein stimulated the expression of anti-apoptotic genes and reduced the pro-apoptotic and pro-inflammatory gene products in chondrocytes. Baicalein promoted SOX-9 expression in a time-dependent manner in chondrocytes. Baicalein inhibited the NF-κB activation that was induced by IL-1β in a time-dependent manner in chondrocytes. Our results suggest that the anti-inflammatory and anti-apoptotic effects of baicalein are mediated through the inhibition of the translocation of phosphorylated p65 to the nucleus.

// Zhaoduan Liang 1 , Ye Tian 1 , Wenxuan Cai 1 , Zhiming Weng 3 , Yanyan Li 1 , Huanling Zhang 1,2 , Yifeng Bao 1 and Yi Li 1,3 1 State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China 2 School of Life Sciences, University of Science and Technology of China, Hefei, China 3 XiangXue Life Sciences Research Center, XiangXue Pharmaceutical Co. Ltd., Guangzhou, China Correspondence to: Yi Li, email: // Keywords : high affinity, programmed cell death protein 1 (PD-1), PD-ligand 1 (PD-L1), soluble PD-L1, T cell, Immunology and Microbiology Section, Immune response, Immunity Received : August 15, 2017 Accepted : September 03, 2017 Published : October 10, 2017 Abstract The activated T cells can be suppressed by programed death-1 (PD-1) axis through low affinity interaction between PD-1 and PD-ligand 1 (PD-L1) in solution or on antigen presenting cells. In clinic, the concentration of soluble PD-L1 in peripheral blood negatively correlates with cancer prognosis. However, there is little information about the relation between the affinity of PD-1/PD-L1 interaction and the suppressive capacity of PD-1 axis. In this study, we analyzed inhibitory roles of high affinity soluble human PD-L1 (hPD-L1) variants, which were generated with directed molecular evolution. Resultant two clones L3C7-hPD-L1 and L3B3-hPD-L1 showed over 20 folds greater affinity than that of native hPD-L1. We found that L3B3-hPD-L1 and L3C7-hPD-L1 could compete with an anti-PD-1 antibody (EH12.1) for binding to hPD-1. More importantly, although native soluble hPD-L1 can induce suppressive effects on activated T cells, we found L3B3-hPD-L1 and L3C7-hPD-L1 attenuated the strength of PD-1 axis for suppressing the proliferation and interferon γ (IFN-γ) secretion of PBMC. In conclusion, our data provide direct evidence in which immune checkpoint receptor-ligand interactive strength can alter the the suppressive function, in particular, the suppressive capacity of PD-1 axis could be decreased with enhanced affinity of soluble PD-L1 and PD-1 interaction. Our study might provide a new direction for manipulating immune checkpoints.

// Xiaolu Huang 1, * , Yiwen Zhou 1, * , Wenhui Liu 1 , Haizhou Li 1 , Xiao Liang 1 , Rui Jin 1 , Hengyu Du 1 , Jizhou He 1 , Bangda Chai 1 , Ran Duan 1 and Qingfeng Li 1 1 Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R.China * These authors contributed equally to this work and should be considered as co-author Correspondence to: Qingfeng Li, email: dr.liqingfeng@shsmu.edu.cn Keywords: silicone implant, autoimmunity, microarray, WGCNA, hub genes Received: July 04, 2017 Accepted: September 21, 2017 Published: October 06, 2017 ABSTRACT Silicone implants are used widely in the field of plastic surgery and are used in a large population. However, their safety profile, especially the silicone-induced immune response, has been a major concern for plastic surgeons for decades. It has been hypothesized that there is a cause and effect relation between silicone and immunity, but this is controversial. The objective of the present study was to determine the hub genes and key pathways related to silicone implant–induced immune responses in a rat model. In addition to cluster and enrichment analyses, we used weighted gene co-expression network analysis (WGCNA) to examine the gene expression profiles in a systematic context. A total five genes ( Fes , Aif1 , Gata3 , Tlr6 , Tlr2 ) were identified as hub genes that are most likely related to the silicone-induced immune response, four of which ( Aif1 , Gata3 , Tlr6 , Tlr2 ) have been associated with autoimmunity as target genes or disease markers. The Toll-like receptor signaling pathway ( p < 0.01, fold enrichment: 7.01) and systemic lupus erythematosus signaling pathway ( p < 0.05, fold enrichment: 5.01), which are considered strongly associated with autoimmunity, were significantly enriched in the silicone-implanted skin samples. The results indicate that silicone implants might trigger the localized immune response, as various immune reaction genes were detected after silicone implantation. The identified five hub genes will hopefully serve as novel therapeutic targets for silicone-related complications and the associated autoimmune diseases.

// 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.

// Kyung-Ah Cho 1 , Minhwa Park 1 , Yu-Hee Kim 1 , Kyung-Ha Ryu 2 and So-Youn Woo 1 1 Department of Microbiology, College of Medicine, Ewha Womans University, Seoul, Republic of Korea 2 Department of Pediatrics, College of Medicine, Ewha Womans University, Seoul, Republic of Korea Correspondence to: So-Youn Woo, email: // Keywords : mesenchymal stem cells, osteoprotegerin, Th17 cells, osteoclasts, psoriasis, Immunology and Microbiology Section, Immune response, Immunity Received : February 20, 2017 Accepted : September 05, 2017 Published : September 28, 2017 Abstract Th17 cells play a critical role in several autoimmune diseases, including psoriasis and psoriatic arthritis (PsA). Psoriasis is a chronic inflammatory skin disease associated with systemic inflammation and comorbidities, such as PsA. PsA develops in nearly 70% of patients with psoriasis, and osteoclasts associated bone erosion is a hallmark of the disease. Thus far, the effect of Th17 cells on osteoclastogenesis via direct cell-to-cell interactions is less understood. In this study, we observed that Th17 cells directly promote osteoclast differentiation and maturation via expression of receptor activator of nuclear factor-κ β ligand (RANKL) in vitro . We investigated the impact of conditioned medium obtained from human palatine tonsil-derived mesenchymal stem cells (T-CM) on the interactions between osteoclasts and Th17 cells. T-CM effectively blunted the RANK-RANKL interaction between the osteoclast precursor cell line RAW 264.7 and Th17 cells via osteoprotegerin (OPG) activity. The frequency of tartrate-resistant acid phosphatase (TRAP)-positive cells in the bone marrow of an imiquimod (IMQ)-induced psoriasis mouse model was decreased following T-CM injection. Therefore, our data provide novel insight into the therapeutic potential of tonsil-derived mesenchymal stem cell-mediated therapy ( via OPG production) for the treatment of pathophysiologic processes induced by osteoclasts under chronic inflammatory conditions such as psoriasis.

// Danlin Yao 1,2,* , Ling Xu 1,2,* , Jiaxiong Tan 1,2 , Yikai Zhang 1,2 , Shuai Lu 1 , Mingde Li 1,2 , Sichun Lu 1,2 , Lijian Yang 1 , Shaohua Chen 1 , Jie Chen 2 , Jing Lai 2 , Yuhong Lu 2 , Xiuli Wu 1,2 , Xianfeng Zha 3 and Yangqiu Li 1,2 1 Key Laboratory for Regenerative Medicine of Ministry of Education, Institute of Hematology, Jinan University, School of Medicine, Jinan University, Guangzhou, China 2 Department of Hematology, First Affiliated Hospital, Jinan University, Guangzhou, China 3 Department of Clinical Laboratory, First Affiliated Hospital, Jinan University, Guangzhou, China * These authors have contributed equally to this work Correspondence to: Yangqiu Li, email: // Keywords : memory T cells, TSCM, CML, TKI, Immunology and Microbiology Section, Immune response, Immunity Received : June 28, 2017 Accepted : August 27, 2017 Published : September 16, 2017 Abstract T cell immune surveillance is considered an important host protection process for inhibiting carcinogenesis. The full capacity of T cell immune surveillance is dependent on T cell homeostasis, particularly for central memory T (T CM ) cells and stem cell memory T (T SCM ) cells. In this study, distribution of T cell subsets in peripheral blood from 12 patients with chronic myeloid leukemia (CML) and 12 cases with CML in complete remission (CR) was analyzed using a multicolor flow cytometer, and 16 samples from healthy individuals (HIs) served as control. The proportion of CD8 + T SCM and CD4 + and CD8 + T CM cells were lower, while CD4 + effector memory T (T EM ) cells and CD4 + and CD8 + terminal effector T (T EF ) cells were higher in CML patients compared with HIs. Moreover, the proportion of CD8 + CD28 - T cells, which were found to have the immune suppressive function, increased in the naive T (T N ) cell and T CM subsets in CML patients compared with HIs. Our study reveals that elimination of leukemia cells by treating with tyrosine kinase inhibitors (TKIs) restores the memory T cell distribution from a skewed pattern in CML patients who are under leukemia burden, indicating that leukemia-specific immune responses mediated by T cells might be induced and maintained in CML patients, however, these responsive T cells might gradually become exhausted due to the continued existence of leukemia cells and their environment; therefore, T cell activation using a different approach remains a key point for enhancing global T cell immunity in CML patients, even for those with CR status.

// Parvin Forghani 1,* , Christopher T. Petersen 1,* and Edmund K. Waller 1 1 Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University, Atlanta, Georgia, USA * These authors have contributed equally to this work Correspondence to: Parvin Forghani, email: // Edmund K. Waller, email: // Keywords : CMV, myeloid-derived suppressor cells (MDSC), VIP, mice, Immunology and Microbiology Section, Immune response, Immunity Received : June 01, 2017 Accepted : August 14, 2017 Published : September 07, 2017 Abstract Vasoactive intestinal peptide (VIP) is recognized as a potent anti-inflammatory factor which affects both the innate and adaptive arms of the immune system. These effects include, but are not limited to, inhibition of T cell proliferation and disruption of immune homeostasis. Myeloid-derived suppressor cells (MDSC) are an immune regulatory cell type that has been described in settings of cancer and infectious disease.Here we demonstrate a reduced circulating monocytic MDSCs in the VIP -/- vs. wild type MCMV. VIP-/- MDSCs secretes less NO upon stimulation with LPS and interferon that relatively lose the ability to suppress T cells activation in vitro compared to wild type MDSCs.Considering the importance of VIP in immunomodulation, the possible effect of VIP in the suppressive function of MDSC populations following CMV infection remains unknown. We describe the possible role of VIP in the regulation of anti-CMV activity of T cells through the activation of MDSCs.

// Yun-Jung Choi 1 , EunSu Kim 1 , Abu Musa Md Talimur Reza 1 , Kwonho Hong 1 , Hyuk Song 1 , Chankyu Park 1 , Seong-Keun Cho 2 , Kiho Lee 3 , Randall S. Prather 4 and Jin-Hoi Kim 1 1 Department of Stem Cell and Regenerative Biotechnology, Humanized Pig Research Center (SRC), Konkuk University, Seoul, Republic of Korea 2 Department of Animal Science, Pusan National University, Miryang, Gyeongnam, Republic of Korea 3 Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, VA, USA 4 Division of Animal Science, University of Missouri-Columbia, Columbia, MO, USA Correspondence to: Jin-Hoi Kim, email: // Keywords : severe combined immunodeficient, SCID, recombination activating gene-2, RAG-2, Talen, Immunology and Microbiology Section, Immune response, Immunity Received : June 22, 2017 Accepted : August 23, 2017 Published : September 02, 2017 Abstract This study comparatively investigated the transcriptional, physiological, and phenotypic differences of the immune disorder between severe combined immunodeficient (SCID) mouse and pig models. We discovered that the recombination activating gene-2 ( Rag-2 ) SCID mice, but not RAG-2 SCID pigs, showed intense, infrequent, and mild cluster of CD3 + -, CD4 + -, and CD8 + signals respectively, suggesting that distinct species-specific effects exist. Furthermore, the expression of six relevant genes ( NFATC1 , CD79B , CD2 , BLNK , FOXO1 , and CD40) was more downregulated than that in the Rag-2 SCID mice, which provides a partial rationale for the death of T/B cells in the lymphoid organs of RAG-2 SCID pigs but not in Rag-2 SCID mice. Further, NK cell maturation-related gene expression was significantly lower in RAG-2 SCID pigs than in Rag-2 SCID mice. Consistently, the RAG-2 SCID pigs, but not Rag-2 SCID mice, developed human induced pluripotent stem cell-derived teratomas that were the same as those of perforin / Rag-2 SCID mice. Therefore, these unexpected findings indicate the superiority of RAG-2 SCID pigs over Rag-2 SCID mice as a suitable model for investigating human diseases.

IFITM3 is involved in cell adhesion, apoptosis, immune, and antivirus activity. Furthermore, IFITM3 gene has been considered as a preferential marker for inflammatory diseases, and positive correlation to pathological grades. Therefore, we assumed that IFITM3 was regulated by different signal pathways. To better understand IFITM3 function in inflammatory response, we cloned swine IFITM3 gene, and detected IFITM3 distribution in tissues, as well as characterized this gene. Results indicated that the length of swine IFITM3 gene was 438 bp, encoding 145 amino acids. IFITM3 gene expression abundance was higher in spleen and lungs. Moreover, we next constructed the eukaryotic expression vector PBIFM3 and transfected into PK15 cells, finally obtained swine IFITM3 gene stable expression cell line. Meanwhile, we explored the effects of LPS on swine IFITM3 expression. Results showed that LPS increased IFITM3 mRNA abundance and exhibited time-dependent effect for LPS treatment. To further demonstrate the mechanism that IFITM3 regulated type I IFNs production, we also detected the important molecules expression of TLR4 signaling pathway. In transfected and non-transfected IFITM3 PK15 cells, LPS exacerbated the relative expression of TLR4-NFκB signaling molecules. However, the IFITM3 overexpression suppressed the inflammatory development of PK15 cells. In conclusion, these data indicated that the overexpression of swine IFITM3 could decrease the inflammatory response through TLR4 signaling pathway, and participate in type I interferon production. These findings may lead to an improved understanding of the biological function of IFITM3 in inflammation.

// Liu Zhenjiang 1 , Martin Rao 1 , Thomas Poiret 1,2 , Silvia Nava 1 , Qingda Meng 1 , Anna von Landenberg 1 , Jiri Bartek Jr. 3,4,5 , Shanshan Xie 6 , Georges Sinclair 3 , Inti Peredo 3 , Ernest Dodoo 3,* and Markus Maeurer 1,2,* 1 Department of Laboratory Medicine, Division of Therapeutic Immunology, Karolinska Institutet, Stockholm, Sweden 2 Centre for Allogeneic Stem Cell Transplantation, Karolinska University Hospital Huddinge, Stockholm, Sweden 3 Department of Neurosurgery, Karolinska University Hospital, Stockholm, Sweden 4 Department of Clinical Neuroscience and Department of Medicine, Karolinska Institutet, Stockholm, Sweden 5 Department of Neurosurgery, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark 6 Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden * These authors have contributed equally to this work Correspondence to: Markus Maeurer, email: // Keywords : glioblastoma, mesothelin, T-cell response, interferon gamma, immunotherapy, Immunology and Microbiology Section, Immune response, Immunity Received : February 25, 2017 Accepted : June 11, 2017 Published : August 16, 2017 Abstract Glioblastoma multiforme (GBM) presents the most malignant form of glioma, with a 5-year survival rate below 3% despite standard therapy. Novel immune-based therapies in improving treatment outcomes in GBM are therefore warranted. Several molecularly defined targets have been identified mediating anti-GBM cellular immune responses. Mesothelin is a tumor-associated antigen (TAA) which is expressed in several solid tumors with different histology. Here, we report the immunological significance of mesothelin in human malignant glioma. Expression of mature, surface-bound mesothelin protein was found to bein human GBM defined by immunofluorescence microscopy, and on freshly isolated, single cell suspension of GBM tumor cells and GBM tumor cell lines, determined by based on flow cytometric analysis. Peripheral blood (PB) from patients with GBM, stimulated with mesothelin peptides and IL-2, IL-15 and IL-21, exhibited increased antigen-specific IFN-γ and TNF-α production. Anti-mesothelin directed T-cell responses could also be detected in tumor - infiltrating lymphocytes (TIL) isolated from GBM speciments. Furthermore, T cells cultured in the presence of IL-2, IL-15 and IL-21 displayed enhanced mesothelin-specific CD4+ and CD8+ subset proliferation, based on ELISA and flow cytometric readouts. Mesothelin-specific IgG antibodies as well as (shed) mature mesothelin protein were detected in plasma samples from patients with GBM by indirect ELISA. Finally yet importantly, we identified distinct immune recognition hotspots within the mature mesothelin component, defined by peptide-specific IFN-γ responses from peripheral T-cells from patients with GBM. Mesothelin may therefore qualify as a viable target for immunotherapeutic approaches for patients with GBM.

// Yingyun Fu 1,3,* , Yazhen Li 1,* , Lan Xu 1 , Shengguo Liu 1 , Minlian Wang 3 , Lu Xiao 1 , Song Liu 2 and Yong Dai 2 1 Department of Respiratory and Critical Care Medicine, The Second Medical College of Jinan University, Shenzhen People’s Hospital, Luohu District, Shenzhen, China 2 Clinical Medical Research Center, The Second Clinical Medical College of Jinan University, Shenzhen People’s Hospital, Luohu District, Shenzhen, China 3 Shenzhen Key Laboratory of Respiratoty Disease, Shenzhen, China * These authors have contributed equally to this work Correspondence to: Yingyun Fu, email: // Keywords : sarcoidosis, immune system, CDR3, TCR BV CDR3, PMBCs, Immunology and Microbiology Section, Immune response, Immunity Received : April 08, 2017 Accepted : July 30, 2017 Published : August 09, 2017 Abstract Sarcoidosis is a systemic granulomatous disorder highly related with immune response. The diversity and stability of the immune system could be measured by hypervariable complementarity-determining region 3 (CDR3) segments of the T cell receptor (TCR). Here we used a combination of multiplex PCR and next-generation sequencing to conduct a good quality analysis of the T-cell receptor BV complementarity-determining region 3 (TCR BV CDR3) gene in peripheral blood mononuclear cells (PBMCs) from 7 sarcoidosis patients and lung sarcoidosis tissue from 6 patients. The length distribution of CDR3 sequences identified a significant difference among CD4+, CD8+ and tissue samples. The analysis of Gini coefficient, Shannon entropy and HEC number showed that they all presents in sarcoidosis tissue group clones in a more skewed manner than that of in PMBCs groups. 2 nucleotide sequences and 2 amino acid sequences were shared by all samples. The comparison of TRBV, TRBJ usage and VJ combination frequency identified 2 TRBV genes, 2 TRBJ genes differentially expressed among different groups and different higher usage and lower usage of V-J combinations between each group.

// Zongwei Lin 1,* , Shao Qian 3,* , Yan Gong 2,* , Jianwei Ren 6 , Lixia Zhao 4 , Dongxiao Wang 5 , Xiaowei Wang 1 , Yun Zhang 1 , Zhe Wang 2 and Qunye Zhang 1 1 The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, and The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Qilu Hospital of Shandong University, Jinan, China 2 Division of Endocrinology and Metabolism, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China 3 Department of Women Health Care, Jinan Maternity and Child Care Hospital, Jinan, China 4 Pharmacy Department of Shandong Medical College, Jinan, China 5 Department of Pharmacy, Chinese PLA General Hospital, Beijing, China 6 Health Division of Guard Bureau, General Staff Department of Chinese PLA, Beijing, China * These authors have equally contributed to this study Correspondence to: Qunye Zhang, email: // Zhe Wang, email: // Keywords : T cell receptor β, atherosclerosis, immune repertoire, next-generation DNA sequencing, complement determining region 3, Immunology and Microbiology Section, Immune response, Immunity Received : April 12, 2017 Accepted : June 18, 2017 Published : August 03, 2017 Abstract The T cell receptor (TCR) β repertoire directly reflects the status of T cell function. Meanwhile, the immune/inflammatory responses regulated by T cells are the critical determinants of atherosclerosis development. However, due to technical limitations, the composition and molecular characteristics of the TCR repertoire in atherosclerotic patients have not been fully elucidated. In the present study, we use powerful immune repertoire sequencing technology to study this issue. Results show that the utilization of V and/or J genes and the diversity of TCRβ repertoire in atherosclerotic plaques are significantly reduced compared to those in the peripheral blood of normal subjects and atherosclerotic patients. The frequencies of the common T cell clones with certain lengths of the complement determining region 3 regions are notably different among all groups. The high-frequency common clones are also increased in the atherosclerotic plaques compared to that in the other two groups. The expansion of several T cell clonotypes (V29-1J2-1, V20-1J1-6, V6-3J2-7 and V11-2J2-2) is validated in atherosclerotic patients. In short, this study reveals that the diversity of TCR β repertoire significantly decreases in atherosclerotic plaques, probably because of the reduced utilization of VJ genes and marked expansion of some T cell subclones. It provides the basis for understanding the roles of T lymphocytes in the pathogenesis of atherosclerosis.

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.

// Young-Jun Kim 1,* , Jae Min Shin 1,* , Su-Hyun Shin 1 , Joo Heon Kim 2 , Ki-Young Sohn 3 , Heung-Jae Kim 3 , Jong-Koo Kang 4 , Sun Young Yoon 3 and Jae Wha Kim 1 1 Cell Factory Research Center, Division of Systems Biology and Bioengineering, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea 2 Department of Pathology, EulJi University School of Medicine, Daejeon, Republic of Korea 3 Enzychem Lifesciences, Daejeon, Republic of Korea 4 Department of Laboratory Animal medicine, College of Veterinary medicine, Chungbuk National University, Cheongju, Republic of Korea * These authors have contributed equally to this work Correspondence to: Jae Wha Kim, email: // Sun Young Yoon, email: // Keywords : PLAG, collagen-induced arthritis, IL-6, STAT3, neutrophil, Immunology and Microbiology Section, Immune response, Immunity Received : December 05, 2016 Accepted : July 06, 2017 Published : July 19, 2017 Abstract The pathogenesis of rheumatoid arthritis (RA) has been implicated neutrophil extracellular traps (NETs) formation which could generate autoantigen. Neutrophil contributes to initiate and maintain the inflammatory process in the joint. In this study, we show that 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) decreases neutrophil migration by regulating the activity of STAT3, a regulator of IL-6 and MIP-2 expression. PLAG caused a decrease in IL-6 production in the RAW264.7 macrophage cell line and in rheumatoid arthritis–fibroblast-like synoviocytes via the regulation of STAT3 signaling without affecting NF-κB signaling. In a mouse model of collagen-induced arthritis (CIA), arthritic symptoms were recapitulated, with increased IL-6 level in the synovium, and PLAG treatment restored IL-6 to a level comparable to that achieved with commercial therapeutics (such as Remicade or methotrexate). Staining of joint tissue with neutrophil-specific antibody showed that PLAG significantly reduced the infiltration of neutrophils into the joint synovium of CIA mice. The inhibitory effect of PLAG on IL-6/STAT3 or MIP-2 signaling also reduced the migration of differentiated neutrophils in vitro . Therefore, PLAG inhibits the infiltration of destructive neutrophils into inflammatory sites, and can be utilized as a potent therapeutic agent for the treatment of sustained inflammation and joint destruction.

// Shouyi Wang 1 , Pin Liu 1 , Pu Yang 1 , Junwen Zheng 1 and Dongchi Zhao 1 1 Department of Pediatrics, Children’s Digital Health and Data Center, Zhongnan Hospital of Wuhan University, Wuhan, China Correspondence to: Dongchi Zhao, email: // Keywords : respiratory syncytial virus, peripheral blood, miRNA, infants, inflammatory response, Immunology and Microbiology Section, Immune response, Immunity Received : July 10, 2016 Accepted : June 27, 2017 Published : July 18, 2017 Abstract MicroRNAs respond to the inflammatory responses induced by RNA virus infection. In this study, we investigated the specific microRNA profile in the peripheral blood of infants infected with respiratory syncytial virus (RSV). Blood specimens were analyzed using microRNA microarrays, followed by quantitative RT-PCR. A specific microRNA profile in the peripheral blood of RSV-infected infants was identified for the first time. MiR-106b-5p, miR-20b-5p, and miR-342-3p were upregulated, while miR-320e, miR-320d, miR-877-5p, miR-122-5p, and miR-92b-5p were downregulated. Pathway analysis indicated that the dysregulated microRNAs were involved in inflammatory and immune responses, including Wnt, TGF-β, insulin, and T and B cell receptor signaling. These results demonstrate that RSV infection associates with a distinct microRNA fingerprint and suggest that RSV induces inflammatory responses in infants.

// 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.

// Haichong Wu 1 , Tao Zhang 1 , Xiaofei Ma 1 , Kangfeng Jiang 1 , Gan Zhao 1 , Changwei Qiu 1 and Ganzhen Deng 1 1 Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, People’s Republic of China Correspondence to: Ganzhen Deng, email: ganzhendeng@sohu.com Keywords: microRNA, bovine endometrial epithelial cell, IFN-τ, pregnancy immunity Received: March 16, 2017 Accepted: May 14, 2017 Published: June 14, 2017 ABSTRACT IFN-τ is specifically secreted by the conceptus in ruminants during early pregnancy, and it plays a vital role in the immunological function of pregnancy. However, its mechanism involving microRNA (miRNA) is still not well understood. Deep sequencing was used to explore the specific miRNA library of IFN-τ on bovine endometrial epithelial cells (bEECs). The results showed that 574 known bovine miRNAs and 109 novel miRNAs were identified. We found 74 differentially expressed miRNAs, including 30 commonly expressed miRNAs in the experiment. Then, qPCR verification of six selected miRNAs showed that they corresponded with the sequencing data. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed significant enrichment of predicted target genes of differentially expressed miRNAs, including influenza A, herpes simplex infection, antigen processing and presentation, viral myocarditis, TNF signaling pathway, graft-versus-host disease, and allograft rejection. These results may provide important contributions to the immune response during early pregnancy in ruminants, but further studies are need to verify the proposed cellular/immunological effects and role of specific miRNA as biomarkers in vivo .

// Mei Chao 1,2,3 , Tzu-Chi Wang 2,* , Chia-Chi Lin 2 , Robert Yung-Liang Wang 2,4 , Wen-Bin Lin 2 , Shang-En Lee 2 , Ying-Yu Cheng 2 , Chau-Ting Yeh 3 and Shan-Bei Iang 2 1 Department of Microbiology and Immunology, Chang Gung University, Guishan, Taoyang, Taiwan 2 Division of Microbiology, Graduate Institute of Biomedical Sciences, Chang Gung University, Guishan, Taoyang, Taiwan 3 Department of Hepato-Gastroenterology, Liver Research Center, Chang Gung Memorial Hospital, Guishan, Taoyang, Taiwan 4 Department of Biomedical Sciences, Chang Gung University, Guishan, Taoyang, Taiwan * Deceased Correspondence to: Mei Chao, email: // Keywords : hepatitis delta virus, mammalian RNA polymerase, ribozyme, RNA recombination, template-switching, Immunology and Microbiology Section, Immune response, Immunity Received : November 09, 2016 Accepted : May 22, 2017 Published : June 01, 2017 Abstract The genome of hepatitis delta virus (HDV) is a 1.7-kb single-stranded circular RNA that folds into an unbranched rod-like structure and has ribozyme activity. HDV redirects host RNA polymerase(s) (RNAP) to perform viral RNA-directed RNA transcription. RNA recombination is known to contribute to the genetic heterogeneity of HDV, but its molecular mechanism is poorly understood. Here, we established a whole-genome HDV-1/HDV-4 recombination map using two cloned sequences coexisting in cultured cells. Our functional analyses of the resulting chimeric delta antigens (the only viral-encoded protein) and recombinant genomes provide insights into how recombination promotes the genotypic and phenotypic diversity of HDV. Our examination of crossover distribution and subsequent mutagenesis analyses demonstrated that ribozyme activity on HDV genome, which is required for viral replication, also contributes to the generation of an inter-clade junction. These data provide circumstantial evidence supporting our contention that HDV RNA recombination occurs via a replication-dependent mechanism. Furthermore, we identify an intrinsic asymmetric bulge on the HDV genome, which appears to promote recombination events in the vicinity. We therefore propose a mammalian RNAP-driven and viral-RNA-structure-promoted template-switching mechanism for HDV genetic recombination. The present findings improve our understanding of the capacities of the host RNAP beyond typical DNA-directed transcription.

// 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.

// Jeong-Eun Lee 1,2,* , Jung-Man Lee 3,* , Young-Jun Park 1 , Byung-Seok Kim 1,2 , Young-Tae Jeon 4,5 and Yeonseok Chung 1,2 1 Laboratory of Immune Regulation, Institute of Pharmaceutical Sciences and College of Pharmacy, Seoul National University, Seoul, Republic of Korea 2 BK21 plus program, College of Pharmacy, Seoul National University, Seoul, Republic of Korea 3 Department of Anesthesiology and Pain Medicine, SMG-SNU Boramae Medical Center, Seoul, Republic of Korea 4 Department of Anesthesiology and Pain Medicine, College of Medicine, Seoul National University, Seoul, Republic of Korea 5 Department of Anesthesiology and Pain Medicine, College of Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea * These authors have equally contributed to this work Correspondence to: Yeonseok Chung, email: // Young-Tae Jeon, email: // Keywords : ketamine, Th17 cell, STAT3, IL-21, autoimmunity, Immunology and Microbiology Section, Immune response, Immunity Received : April 04, 2017 Accepted : May 23, 2017 Published : May 31, 2017 Abstract Ketamine is widely used in animals and humans as a systemic anesthetic. Although several immune-modulatory functions of ketamine have been reported, the effects of ketamine on the differentiation of Th17 cell are unknown. We found that ketamine significantly diminished the frequency of IL-17-producers among CD4 + T cells stimulated under Th17-skewing conditions. Mechanistic studies showed that ketamine had little effect on the production of Th17-inducing cytokines by dendritic cells and the proliferation of T cells in response to anti-CD3; however it significantly hampered IL-21 expression as well as STAT3 phosphorylation in T cells upon IL-6 stimulation. Moreover, MOG-reactive CD4 + T cells expanded in the presence of ketamine produced reduced amounts of Th17 cytokines, leading to diminished EAE severity when transferred into TCRβ-deficient mice in comparison to those treated with vehicle. These findings demonstrate that ketamine suppresses autoimmune Th17 cell responses by inhibiting the differentiation as well as the reactivation of Th17 cells.

// Pinelopi Samara 1 , Vivi Miriagou 2 , Michael Zachariadis 1,3 , Olga Mavrofrydi 1 , Vasilis J. Promponas 4 , Skarlatos G. Dedos 1 , Panagiota Papazafiri 1 , Hubert Kalbacher 5 , Wolfgang Voelter 5 and Ourania Tsitsilonis 1 1 Department of Biology, National and Kapodistrian University of Athens, Athens, Greece 2 Laboratory of Bacteriology, Hellenic Pasteur Institute, Athens, Greece 3 Institute of Biosciences and Applications, National Center for Scientific Research “Demokritos”, Agia Paraskevi, Athens, Greece 4 Bioinformatics Research Laboratory, Department of Biological Sciences, University of Cyprus, Nicosia, Cyprus 5 Interfaculty Institute of Biochemistry, University of Tubingen, Tubingen, Germany Correspondence to: Ourania Tsitsilonis, email: // Pinelopi Samara, email: // Keywords : apoptosis, biomarker, Klebsiella pneumoniae, proTα(100-109), sepsis, Immunology and Microbiology Section, Immune response, Immunity Received : December 15, 2016 Accepted : April 17, 2017 Published : May 24, 2017 Abstract Sepsis is a life-threatening condition that requires urgent care. Thus, the identification of specific and sensitive biomarkers for its early diagnosis and management are of clinical importance. The alarmin prothymosin alpha (proTα) and its decapeptide proTα(100-109) are immunostimulatory peptides related to cell death. In this study, we generated bacterial models of sepsis in mice using two Klebsiella pneumoniae strains (L-78 and ATCC 43816) and monitored sepsis progression using proTα(100-109) as a biomarker. Serum concentration of proTα(100-109) gradually increased as sepsis progressed in mice infected with L-78, a strain which, unlike ATCC 43816, was phagocytosed by monocytes/macrophages. Analysis of splenocytes from L-78-infected animals revealed that post-infection spleen monocytes/macrophages were gradually driven to caspase-3-mediated apoptosis. These results were verified in vitro in L-78-infected human monocytes/macrophages. Efficient phagocytosis of L-78 by monocytes stimulated their apoptosis and the concentration of proTα(100-109) in culture supernatants increased. Human macrophages strongly phagocytosed L-78, but resisted cell death. This is the first report suggesting that high levels of proTα(100-109) correlate, both in vitro and in vivo , with increased percentages of cell apoptosis. Moreover, we showed that low levels of proTα(100-109) early post-infection likely correlate with sepsis resolution and thus, the decapeptide could eventually serve as an early surrogate biomarker for predicting bacteria-induced sepsis outcome.

// Di Li 1,2,* , Ya Yang 1,* , Zhiqiang Tian 3 , Jun Lv 1 , Fengjun Sun 1 , Qian Wang 1 , Yao Liu 1 and Peiyuan Xia 1 1 Department of Pharmacy, Southwest Hospital, Third Military Medical University, Chongqing, China 2 Department of Pharmacy, Second Affiliated Hospital, Chongqing Medical University, Chongqing, China 3 Department of Immunology, Third Military Medical University, Chongqing, China * These authors have contributed equally to this work Correspondence to: Peiyuan Xia, email: // Yao Liu, email: // Keywords : antimicrobial peptides, CLP-19, synergistic effect, hydroxyl radicals, LPS, Immunology and Microbiology Section, Immune response, Immunity Received : January 17, 2017 Accepted :April 15, 2017 Published : May 23, 2017 Abstract The treatment of drug-resistant infections is complicated and the alarming rise in infectious diseases poses a unique challenge for development of effective therapeutic strategies. Antibiotic-induced liberation of the bacterial endotoxin lipopolysaccharide (LPS) may have immediate adverse effects promoting septic shock in patients. In the present study, we first confirmed our previous finding that looped antimicrobial peptide CLP-19 exerts non-specific direct antibacterial activity with no toxic to mammalian cells and second revealed that CLP-19 has synergistic effect to enhance the antibacterial activities of other conventional bactericidal (ampicillin and ceftazidime) and bacteriostatic (erythromycin and levofloxacin) agents. Third, the underlying mechanism of antibiotic effect was likely associated with stimulation of hydroxyl radical generation. Lastly, CLP-19 was shown to effectively reduce the antibiotic-induced liberation of LPS, through direct neutralization of the LPS. Thus, CLP-19 is a potential therapeutic agent for combinatorial antibiotic therapy.

// Yi-Zhong Wu 1, 2, * , Gao Tan 1, * , Fang Wu 3 and Fa-Chao Zhi 1 1 Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, China 2 Department of Gastroenterology, Hunan Provincial People’s Hospital, Changsha, China 3 Department of Gastroenterology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China * These authors contributed equally to this work Correspondence to: Fa-Chao Zhi, email: zhifc41532@163.com Gao Tan, email: tgao0316@163.com Keywords: H. pylori, Crohn’s disease, mucosal immunology, Th cells Received: September 26, 2016 Accepted: April 19, 2017 Published: May 18, 2017 ABSTRACT There is an epidemiological inverse relationship between Helicobacter pylori ( H. pylori ) infection and Crohn’s disease (CD). However, whether H. pylori plays a protective role against CD remains unclear. Since 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis is thought to resemble CD, we investigated whether H. pylori can attenuate TNBS-induced colitis in mice. Here we show that H. pylori can attenuate the severity of TNBS-induced colitis. In addition, H. pylori not only down-regulates Th17 and Th1 cytokine expression, but can up-regulate Th2 cytokine expression and increase the Th2:Th17 ratio of CD4 + T in the colonic mucosa of TNBS-induced colitis. Our results indicate that H. pylori attenuates TNBS-induced colitis mainly through increasing Th2 cells in murine colonic mucosa. Our finding offers a novel view on the role of H. pylori in regulating gastrointestinal immunity, and may open a new avenue for development of therapeutic strategies in CD by making use of asymptomatic H. pylori colonization.

// Matthias Zimmermann 1 , Lucian Beer 1,2 , Robert Ullrich 3 , Dominika Lukovic 4 , Elisabeth Simader 1 , Denise Traxler 1,4 , Tanja Wagner 5 , Lucas Nemec 5 , Lukas Altenburger 5 , Andreas Zuckermann 6 , Mariann Gyongyosi 4 , Hendrik Jan Ankersmit 1,5 and Michael Mildner 7 1 Christian Doppler Laboratory for Cardiac and Thoracic Diagnosis and Regeneration, Medical University of Vienna, Vienna, Austria 2 Department of Biomedical Imaging and Image-Guided Therapy, Medical University of Vienna, Vienna, Austria 3 Department of Pathology, Medical University of Vienna, Vienna, Austria 4 Department of Cardiology, Medical University of Vienna, Vienna, Austria 5 Department of Thoracic Surgery, Medical University of Vienna, Vienna, Austria 6 Department of Cardiac Surgery, Medical University of Vienna, Vienna, Austria 7 Department of Dermatology, Research Division of Biology and Pathobiology of the Skin, Medical University of Vienna, Vienna, Austria Correspondence to: Michael Mildner, email: // Hendrik Jan Ankersmit, email: // Keywords : myocardial infarction, systemic effect, paracrine factors, transcription factor, Klf4, Immunology and Microbiology Section, Immune response, Immunity Received : April 05, 2017 Accepted : May 03, 2017 Published : May 17, 2017 Abstract Aims: Ischemic myocardial injury leads to the activation of inflammatory mechanisms and results in ventricular remodeling. Although great efforts have been made to unravel the molecular and cellular processes taking place in the ischemic myocardium, little is known about the effects on the surrounding tissue and other organs. The aim of this study was to determine region specific differences in the myocardium and in distant organs after experimental myocardial infarction by using a bioinformatics approach. Methods and Results: A porcine closed chest reperfused acute myocardial infarction model and mRNA microarrays have been used to evaluate gene expression changes. Myocardial infarction changed the expression of 8903 genes in myocardial-, 856 in hepatic- and 338 in splenic tissue. Identification of myocardial region specific differences as well as expression profiling of distant organs revealed clear gene-regulation patterns within the first 24 hours after ischemia. Transcription factor binding site analysis suggested a strong role for Kruppel like factor 4 (Klf4) in the regulation of gene expression following myocardial infarction, and was therefore investigated further by immunohistochemistry. Strong nuclear Klf4 expression with clear region specific differences was detectable in porcine and human heart samples after myocardial infarction. Conclusion: Apart from presenting a post myocardial infarction gene expression database and specific response pathways, the key message of this work is that myocardial ischemia does not end at the injured myocardium. The present results have enlarged the spectrum of organs affected, and suggest that a variety of organ systems are involved in the co-ordination of the organism´s response to myocardial infarction.

// Gang Liu 1 , Lei Yu 2 , Jun Fang 3 , Chien-An Andy Hu 1, 4 , Jie Yin 1 , Hengjia Ni 1 , Wenkai Ren 1 , Veeramuthu Duraipandiyan 5 , Shuai Chen 1 , Naif Abdullah Al-Dhabi 5 and Yulong Yin 1, 6, 7 1 Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China 2 China Animal Disease Control Center, Beijing 102618, China 3 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China 4 Department of Biochemistry and Molecular Biology, University of New Mexico School of Medicine, Albuquerque, New Mexico 87131, USA 5 Addiriyah Research Chair for Environmental Studies, Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia 6 College of Animal Science, South China Agricultural University, Guangzhou 510642, China 7 Laboratory of Animal Nutrition and Human Health, School of Life Sciences, Hunan Normal University, Changsha 410081, China Correspondence to: Wenkai Ren, email: renwenkai19@126.com Yulong Yin, email: yinyulong@isa.ac.cn Keywords: inflammatory bowel disease, methionine restriction, diet, oxidative stress, NF-κB Received: December 28, 2016 Accepted: April 14, 2017 Published: May 11, 2017 ABSTRACT A strong correlation exists between inflammatory bowel disease (IBD) and oxidative stress involving alterations of several key signaling pathways. It is known that methionine promotes reactive oxygen species (ROS) production; we therefore hypothesize that a methionine restriction diet would reduce ROS production, inflammatory responses, and the course of IBD. We generated a murine colitis model by dextran sodium sulfate (DSS) treatment and tested the effects of the methionine restriction diet. Forty-eight mice were randomly divided into four groups of equal size, which included a control (CON) group, an MR (methionine restriction diet) group, a DSS treated group and an MR-DSS treated group. Mice in the first two groups had unrestricted access to water for one week. Mice in the two DSS-treated groups had unrestricted access to 5% DSS solution supplied in the drinking water for the same period. Mice in the CON and DSS groups were given a basal diet, whereas mice in the MR-DSS and MR groups were fed a 0.14% MR diet. We found that DSS reduced daily weight gain, suppressed antioxidant enzyme expression, increased histopathology scores and activated NF-κB and nuclear factor erythroid 2-related factor 2/Kelch-like ECH-associated protein 1 (Nrf2/Keap1) signaling. We also showed that the MR diet upregulated catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities, decreased myeloperoxidase (MPO), TNF-α and IL-1β, and reversed activation of the NF-κB signaling pathway in MR-DSS mice. Taken together, our results imply that the MR diet may be considered as an adjuvant in IBD therapeutics.

// Chengcheng Song 1, 2 , Xiu-Jing Zheng 1 , Chang-Cheng Liu 1 , Yifa Zhou 2 and Xin-Shan Ye 1 1 State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China 2 School of Life Sciences, Northeast Normal University, Changchun 130024, China Correspondence to: Xin-Shan Ye, email: xinshan@bjmu.edu.cn Yifa Zhou, email: zhouyf383@nenu.edu.cn Keywords: cancer immunotherapy, glycoconjugate vaccine, tumor-associated carbohydrate antigen, fluoro-substituted STn, cross reaction Received: March 08, 2017 Accepted: April 19, 2017 Published: May 07, 2017 ABSTRACT Development of an effective vaccine to target tumor associated carbohydrate antigens, aberrantly expressed on the cell surface of various carcinomas, is an appealing approach toward cancer immunotherapy. However, a major problem of carbohydrate antigens is their poor immunogenicity. Immunization with modified-carbohydrate antigens could improve the immunogenicity and induce cross reaction with the native carbohydrate antigens. In this study, we investigated the antitumor ability of three fluoro-substituted sialyl-Tn (STn) analogues (2, 3, 4) coupled to KLH (keyhole limpet hemocyanin) and studied the mechanism of tumor immunotherapy of the vaccines in a murine model of colon cancer. Vaccination with 4-KLH, in which the two N -acetyl groups of STn are substituted with N -fluoroacetyl groups, could remarkably prolong the survival of tumor-bearing mouse and resulted in a significant reduction in tumor burden of lungs compared with STn-KLH (1-KLH). The vaccine 4-KLH could provoke stronger cytotoxic T lymphocytes immune response, T helper (Th) cell-mediated immune response and an earlier-stage Th1 immune response than 1-KLH, thus breaking immune tolerance and generating a therapeutic response. The 4-KLH vaccine induced strong tumor-specific anti-STn antibodies which could mediate complement-dependent cytotoxicity and antibody-dependent cell-mediated cytotoxicity against human tumor cells. Moreover, in the absence of adjuvant, 4-KLH still elicited stronger immune responses than 1-KLH. Our data suggested that 4-KLH is superior in tumor prevention. The strategic hapten fluorination may be a potential approach applicable to the vaccines development for the cancer immunotherapy.

// Che-Hsin Lee 1, 2 , Tomoyuki Nishikawa 3 , Yasufumi Kaneda 3 1 Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan 2 Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan 3 Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, Osaka, Japan Correspondence to: Che-Hsin Lee, email: chlee@mail.nsysu.edu.tw Yasufumi Kaneda, email: kaneday@gts.med.osaka-u.ac.jp Keywords: Salmonella, targeted therapy, polymer, hemagglutinating virus of Japan-envelope Received: December 08, 2016 Accepted: March 30, 2017 Published: April 11, 2017 ABSTRACT Salmonella can target to tumor microenvironments after systemic treatment. The hemagglutinating virus of Japan-envelope (HVJ-E) induced apoptosis in tumor cells without toxicity in normal cells. Current HVJ-E therapeutic strategies, aimed at using HVJ-E for intratumor treatment, have shown great promise in animal models but have achieved only limited systemic treatment. The purpose of this study was to investigate the modulation of the anti-tumor efficiency of HVJ-E by coating the particles with poly (allylamine hydrochloride) (PAH), designated as P-HVJ-E. Treatment with P-HVJ-E resulted in decreased hemagglutinating activity and maintained tumor cell-selective apoptosis and anti-tumor immunity. The use of Salmonella as a coating for P-HVJ-E (PHS) enhanced the antitumor activity and maintained the tumor-targeting activity. Treatment with PHS resulted in delayed tumor growth in tumor-bearing mice. Furthermore, a Western blot assay of the tumors revealed that HVJ-E targeted to the tumor after systemic treatment with PHS. These results indicate that Salmonella coating viral particles may provide a new approach for tumor therapy.

// 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.

// Ledan Wang 1 , Yiqi Cai 3 , Yirong Xiong 2 , Wangqi Du 2 , Danwei Cen 2 , Chanqiong Zhang 2 , Yiling Song 2 , Shanli Zhu 2 , Xiangyang Xue 2 , Lifang Zhang 2 1 Department of Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325035, Zhejiang, China 2 Department of Microbiology and Immunology, Institute of Molecular Virology and Immunology, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China 3 Department of Gastrointestinal, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325035, Zhejiang, China Correspondence to: Lifang Zhang, email: wenzhouzlf@126.com Keywords: Chlamydia trachomatis, major outer membrane protein, multi-epitopes, human papillomavirus, vaccine Received: February 06, 2017 Accepted: March 15, 2017 Published: March 27, 2017 ABSTRACT Chlamydia trachomatis ( Ct ) is one of the most frequently encountered sexual infection all over the world, yielding tremendous reproductive problems (e.g. infertility and ectopic pregnancy) in the women. This work described the design of a plasmid vaccine that protect mice from Ct infection, and reduce productive tract damage by generating effective antibody and cytotoxic T cell immunity. The vaccine, s was composed of MOMP multi-epitope and HPV16L2 genes carried in pcDNA plasmid (i.e. pcDNA3.1/MOMP/HPV16L). In transfection, the vaccine expressed the chimeric genes (i.e. MOMP and HPV16L2), as demonstrated via western blot, RT-PCR and fluorescence imaging. In vitro , the vaccine transfected COS-7 cells and expressed the proteins corresponding to the genes carried in the vaccine. Through intramuscular immunization in BALB/c mice, the vaccine induced higher levels of anti- Ct IgG titer, anti-HPV16L2 IgG titer in serum and IgA titer in local mucosal secretions, compared to plasmid vaccines that carry only Ct MOMP multi-epitope or HPV16L2 chimeric component only. In mice intravaginally challenged with Ct , the vaccines pcDNA3.1/MOMP/HPV16L2 generated a higher level of genital protection compared to other vaccine formulations. Additionally, histochemical staining indicated that pcDNA3.1/MOMP/HPV16L2 eliminated mouse genital tract tissue pathologies induced by Ct infection. This work demonstrated that pcDNA/MOMP/HPV16L2 vaccine can protect against Ct infection by regulating antibody production, cytotoxic T cell killing functions and reducing pathological damage in mice genital tract. This work can potentially offer us a new vaccine platform against Ct infection.

Vaccination is a potential approach for prevention and control of disease in fish. The use of genetically engineered vaccines is an effective method and a green intervention to control bacterial infection in aquaculture. However, efforts to develop these vaccines are limited by the lack of conserved protective antigens. In this study, three candidate immunogens (Srr, NeuA, and Hsp) of the pathogenic Streptococcus iniae strain DGX07 isolated from diseased channel catfish were identified and analyzed. Molecular cloning, expression, and purification of candidate antigen genes were carried out to obtain the candidate immunogens in the form of recombinant subunit vaccines. Western blotting was performed to evaluate immunogenicity in vitro and channel catfish were vaccinated by intraperitoneal injection and the specific antibody titers and relative percent of survival were determined to evaluate immune protection in vivo. The results showed that these three candidate immunogens were expressed correctly as recombinant proteins fused with His tags, with molecular weights of 70 kDa for Srr, 86 kDa for NeuA, and 51 kDa for Hsp, respectively. Moreover, each immunogen was predicted to be located either extracellularly or on the surface of S. iniae, and were able to offer protection against S. iniae infection in the form of recombinant subunit vaccines with adjuvant ISA763, especially Srr, with a relative percent of survival of 70% for Srr, 55% for NeuA, and 50% for Hsp, respectively.

// Sabrina Giampaolo 1 , Gabriela Wojcik 2 , Edgar Serfling 1,3 and Amiya K. Patra 1,2 1 Department of Molecular Pathology, Institute of Pathology, University of Wurzburg, Wurzburg, Germany 2 Institute of Translational and Stratified Medicine, Peninsula Schools of Medicine and Dentistry, University of Plymouth, Plymouth, UK 3 Comprehensive Cancer Center Mainfranken, University of Wurzburg, Wurzburg, Germany Correspondence to: Amiya K. Patra, email: // Keywords : hematopoietic stem cells, IL-2, Treg cells, IL-10, IFN-γ, Immunology and Microbiology Section, Immune response, Immunity Received : November 17, 2016 Accepted : March 06, 2017 Published : March 18, 2017 Abstract The role of IL-2 in HSC maintenance is unknown. Here we show that Il2 -/- mice develop severe anomalies in HSC maintenance leading to defective hematopoiesis. Whereas, lack of IL-2 signaling was detrimental for lympho- and erythropoiesis, myelopoiesis was enhanced in Il2 -/- mice. Investigation of the underlying mechanisms of dysregulated hematopoiesis in Il2 -/- mice shows that the IL-2-T reg cell axis is indispensable for HSC maintenance and normal hematopoiesis. Lack of T reg activity resulted in increased IFN-γ production by activated T cells and an expansion of the HSCs in the bone marrow (BM). Though, restoring T reg population successfully rescued HSC maintenance in Il2 -/- mice, preventing IFN-γ activity could do the same even in the absence of T reg cells. Our study suggests that equilibrium in IL-2 and IFN-γ activity is critical for steady state hematopoiesis, and in clinical conditions of BM failure, IL-2 or anti-IFN-γ treatment might help to restore hematopoiesis.

Tec kinase, a prototypical member of the Tec tyrosine kinases family, was shown to mainly govern lymphocyte proliferation. In the present study, we investigated the role of Tec kinase in acute inflammatory response in lipopolysaccharide (LPS) challenge. First, we demonstrate that Tec kinase activity was observed in RAW264.7 macrophages exposed to LPS. Tec and phosphorylated Tec expression were upregulated in a dose- and time-dependent manner after LPS stimulation. LPS increased monocyte chemotactic protein (MCP)-1 secretion and intercellular adhesion molecule (ICAM)-1 expression, and increasing mRNA expression was consistently observed. LPS also induced IκBα phoshporylaytion and its degradation, increased NF-κB p65 phoshporylaytion and translocation to nuclei in RAW264.7 cells. Pretreatment with LFM-A13 decreased LPS-induced cytokines and chemokines production and mRNA levels, blocked NF-κB transactivation. These effects of LPS were also prevented by Tec-siRNA. Additionally, LFM-A13 or Tec-siRNA obviously inhibited LPS-induced TGFβ-activated kinase 1(TAK1) phosphorylation. Taken together, our results suggest that Tec kinase involves in acute inflammation process in LPS-stimulated RAW264.7 cells, at least mediated by activating TAK1/ NF-κB signal pathway.

// Lei Tu 1,* , Jie Chen 2,* , Dandan Xu 2 , Zhongming Xie 2 , Bing Yu 3 , Ying Tao 3 , Guixiu Shi 3 and Lihua Duan 3 1 Division of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China 2 College of Medicine, Xiamen University, Fujian, China 3 Department of Rheumatology and Clinical Immunology, The First Affiliated Hospital of Xiamen University, Xiamen, Fujian, China * These authors have contributed equally to this work Correspondence to: Lihua Duan, email: // Guixiu Shi, email: // Keywords : IL-33, inflammatory bowel disease, macrophage, M2, Arginase-1, Immunology and Microbiology Section, Immune response, Immunity Received : January 17, 2017 Accepted : February 28, 2017 Published : March 07, 2017 Abstract Accumulated data have shown that alternatively activated macrophage exerts a modulatory role in many diseases, including colitis. Interleukin-33 (IL-33), a critical modulator in adaptive and innate immune, has been implicated in autoimmunity and inflammation. Previously, we have reported that IL-33 functions as a protective modulator in TNBS-induced colitis, which is closely related to a Th1-to-Th2/Treg switch. Here, we present novel evidence suggesting that IL-33 primes macrophage into alternatively activated macrophages (AAM) in TNBS-induced colitis. The strong polarized effect of IL-33 was tightly associated with the markedly increased induction of Th2-type cytokines. To confirm the beneficial effects of AAM induced by IL-33, peritoneal AAMs isolated from IL-33-treated mice were transferred to recipient mice with TNBS colitis. The adoptive transfer resulted in prominent inhibition of disease activity and inflammatory cytokines in the TNBS-treated mice. In conclusion, our data provide clear evidence that IL-33 plays a protective role in TNBS-induced colitis, which is closely related to AAM polarization.

// 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.