bluuebunny/crossref_metadata_2025_split huggingface dataset:
Apocynin inhibits reactive oxygen species (ROS) and downregulates chemokine and cytokine production induced by avian influenza H5N1and H7N9 virus infection. (INC8P.433)
Authors: Stambas John , Lowther Sue , Ye Siying 
Published: January 2023

Abstract
Influenza A virus infection causes acute inflammation resulting in increased reactive oxygen species (ROS) production and lung injury. ROS are produced following activation of the NADPH oxidase enzyme complex. Apocynin, an inhibitor of NADPH oxidase complex formation has been shown to reduce the accumulation of inflammatory cells in the lungs of influenza-infected mice. As hypercytokinemia is thought to contribute to severe disease pathogenesis of both H5N1 and H7N9 infection in humans, we hypothesized that apocynin could reduce virus-induced cytokine/chemokine production. Human lung epithelial and chicken cell lines were infected with low pathogenic avian influenza H7N9 (A/Anhui/1/2013) and HPAI H5N1 (A/chicken/Vietnam/0008/2004) viruses in the absence or presence of apocynin. Quantitative real-time PCR demonstrated that apocynin inhibited influenza-induced cytokines/chemokines and ROS production following infection. Interestingly, addition of apocynin to in vitro cultures significantly increased SOCS1 and SOCS3 mRNA and protein expression, contributing to the negative regulation of overall cytokine expression. We suggest that intervention strategies targeting both the virus (current therapeutics) and the host e.g. apocynin (future therapeutics) could be used to ameliorate disease in infected individuals, reducing hypercytokinemia and pathology and allowing full development of adaptive immune responses.
Dendritic cell CD209a expression is critical for the development of pathogenic Th17 cell responses in murine schistosomiasis (INC9P.443)
Authors: Ponichtera Holly , Shainheit Mara , Liu Beiyun , Raychowdhury Raktima , Larkin Bridget , Russo Joanne , Salantes Danielle , Lai Chao-Qiang , Parnell Laurence , Yun Tae , Cheong Cheolho , Bunnell Stephen , Hacohen Nir , Stadecker Miguel 
Published: January 2023

Abstract
The magnitude of immunopathology and pro-inflammatory cytokine production in murine Schistosoma mansoni infection is strain-dependent. Severe hepatic egg-induced granulomatous inflammation in CBA mice is associated with Th1 and Th17 cytokine responses, whereas BL/6 mice develop milder lesions in a Th2-polarized cytokine environment. Pathogenic Th17 cell responses in CBA mice are dependent on the production of IL-1β and IL-23 by egg-stimulated dendritic cells (DC); by comparison, such Th17 cells fail to develop in BL/6 mice. The reasons for strain-dependent differences in DC reactivity to eggs remain unclear. Genome-wide gene profiling revealed significant differences between CBA vs. BL/6 DCs in C-type lectin receptors (CLRs), a family of pattern recognition receptors that binds glycans such as those produced by schistosome eggs. Expression of the CLR CD209a, a murine homologue of human DC-specific ICAM-3-grabbing non-integrin (DC-SIGN), was strikingly higher in several APC populations from CBA mice; however, only CBA DC, but not macrophages, B cells, or granulocytes elicited Th17 cell differentiation in response to schistosome eggs. Gene silencing in CBA DC, and over-expression in BL/6 DC, demonstrated CD209a to be necessary for egg-induced DC production of ERK1/2 map kinase-dependent IL-1β and IL-23 as well as subsequent Th17 cell development. These findings reveal a novel mechanism controlling the development of Th17 cell-mediated immunopathology in helminthic disease.
Inhibition of TLR2 signaling by small molecule inhibitors targeting a novel putative pocket within the TLR2 TIR domain (INM9P.448)
Authors: Mistry Pragnesh , Laird Michelle , Schwarz Ryan , Greene Shannon , Dyson Tristan , Jabre Sandra , Snyder Greg , Deredge Daniel , Chauhan Jamal , Wintrode Patrick , Xiao Tsan , Sundberg Eric , Fletcher Steven , Toshchakov Vladimir , MacKerell Alexander , Vogel Stefanie 
Published: January 2023

Abstract
Overexuberant TLR2 signaling has been implicated in numerous diseases. Visual analysis of the TLR2 TIR domain crystal structure suggested the presence of a “pocket” adjacent to the highly conserved and functionally important proline and glycine residues of the BB loop. Given the importance of the BB loop in mediating TLR signal transduction, we hypothesized that Computer-Aided Drug Design (CADD) could be used to identify a small molecule inhibitor(s) that would fit within this “pocket” and blunt TLR2 signaling. Using CADD, ~150 small compounds were identified based on their predicted ability to bind in this pocket. Based on inhibition of IL-8 mRNA induced by TLR2 agonists, compound “C29” was found to inhibit TLR2/1 and TLR2/6 signaling in human HEK-TLR2 and THP-1 cells, but only TLR2/1 signaling in murine macrophages. C29 blocks heat-killed and live bacterial TLR2 agonist-induced proinflammatory cytokine mRNA. C29 prevents the early activation of TLR2-mediated signaling, including NF-κB and MAPKs, as well as MyD88 recruitment to TLR2 in THP-1 cells. C29 is cleavable and the byproduct, o-vanillin, reproduces comparable TLR2 inhibitory activity. O-vanillin covalently interacts with recombinant human TLR2 TIR domain dose-dependently. Mutagenesis of “pocket” residues revealed an indispensable role for TLR2/1, but not TLR2/6, signaling, suggesting divergent roles. Collectively, these results provide proof-of-principle for using CADD to identify inhibitors of TLR2 signaling.
CD40 ligation by T cells downregulates macrophage TNF production by IRAK1-mediated IL-10 expression and controls innate hyperinflammation (INM9P.462)
Authors: Inoue Makoto , Shinohara Mari 
Published: January 2023

Abstract
Endotoxemia is caused by excessive inflammation, but immune system has various mechanisms to avoid collateral organ damages in endotoxemia. A handful of reports have shown that innate immune responses are suppressed by the adaptive immune system. However, the molecular mechanism by which adaptive immune cells suppress innate inflammatory responses is not clear. Here, we found that T cell-macrophage interaction prolonged survival of mice in lipopolysaccharide (LPS)-induced enodotoxemia through controlling TNF levels, demonstrated by T cell reconstitution in Rag2-deficient recipients. Downregulation of macrophage TNF expression was achieved by crosstalk between CD40 and TLR4 signaling pathways, which mediated IRAK1 nuclear translocation and its binding to the Il10 promoter in macrophages, without interfering the NFκB pathway. IL-10 is then detected by macrophages in an autocrine fashion to destabilize Tnfa mRNA, resulting in attenuated TNF levels. Simultaneous stimulation of CD40 and TLR4 signaling pathways was essential to induce IRAK1-mediated IL-10 expression; because CD40 signaling licenses IRAK1 nuclear translocation by IRAK1 sumoylation in the presence of TRAF2 and intracellular isoform of osteopontin (iOPN), while TLR4 signaling provides IRF5 as a chaperone of sumoylated IRAK1 for its nuclear translocation. Our study demonstrates a mechanistic basis of the immunosuppressive role of macrophage CD40 in LPS endotoxemia.
Molecular mechanisms responsible for the dynamic modulation of macrophage responses to varying dosages of LPS (INM9P.453)
Authors: Morris Matt , Li Liwu 
Published: January 2023

Abstract
Monocytes and macrophages can be dynamically programmed into distinct states depending upon the strength of external stimulants. Innate programming may bear significant relevance to the pathogenesis and resolution of human inflammatory diseases. However, systems analyses of the molecular mechanisms responsible for the dynamic programming of innate leukocytes are lacking. In this study, we focused on the macrophage responses to lipopolysaccharide (LPS), a ubiquitous bacterial endotoxin recognized by Toll-like receptor 4 (TLR4). We employed both immortalized human monocytic THP-1 cells and primary murine bone-marrow derived macrophages (BMDM). We observed that varying doses of LPS induce distinct expression profiles of selected pro- and anti- inflammatory cytokines. Lower doses of LPS preferentially suppressed anti-inflammatory mediators, and selectively induced pro-inflammatory mediators. Rising LPS dosages caused a shift in expression profiles, giving rise to double-positive cells. Higher LPS dosages shifted the landscape to anti-inflammatory mediators and drove down the expression of pro-inflammatory mediators. By utilizing BMDM from IRAK-M knockout mice, we identified IRAK-M as a key player in this process, as IRAK-M deletion skews the dynamic responses to varying dosages of LPS. Our study provides a systems view of macrophage programming, and may shed light on our understanding of the dynamic balance between pathogenesis and resolution of inflammatory diseases.
Mesenchymal stem cells differentially modulate effector CD8+ T cells and exacerbate EAE. (IRC8P.484)
Authors: Glenn Justin , Smith Matthew , Whartenby Katharine , Calabresi Peter 
Published: January 2023

Abstract
Auto-reactive T cells are a significant cause of myelin destruction and eventual axon loss in EAE and probably multiple sclerosis (MS). Both CD4+ and CD8+ T cells acquire pathogenic IFNγ-producing- (Type I) and IL-17A-producing- (Type 17) effector phenotypes in MS and EAE. On the frontier of MS therapy, mesenchymal stem cells (MSC) have emerged as a promising candidate for inflammatory suppression and disease amelioration. Although MSC have been extensively demonstrated to suppress pathogenic effector CD4+ T cells and CD4+ T cell-mediated EAE, surprisingly few studies have addressed their modulation of effector CD8+ T cells in MS or EAE. We find that MSC differentially modulate CD8+ T cell development depending on effector T cell subtype. MSC drive activated low-IFNγ producers toward an enhanced high-IFNγ Tc1-like phenotype but strongly inhibit the production of IL-17A and Tc17 polarization in vitro. These observations are underscored by differential MSC modulation of T cell activation, proliferation, and signature transcription factor up-regulation. In addition, MSC constitutively produced a panel of Type I inflammatory cytokines associated with the IFNγ/IL-17A axis.Based on these in vitro effects on CD8+ T cells, we next evaluated their impact on the severity of EAE. To better evaluate CD8+ T cells, we immunized mice with MOG 37-50, which is a CD8-targeted epitope. Our results revealed a disease worsening, consistent with the in vitro stimulation of Tc1 cells.
Local intravaginal microencapsulated interleukin 12 treatment enhances adaptive immunity to Neisseria gonorrhoeae and induces cross-protection against heterologous strains of gonococci (IRC8P.492)
Authors: Liu Yingru , Phillips Tara , Egilmez Nejat , Russell Michael 
Published: January 2023

Abstract
Uncomplicated genital infection with N. gonorrhoeae does not induce specific protective immunity or immune memory. Our previous studies in a mouse genital tract infection model have demonstrated that N. gonorrhoeae selectively elicits Th17-mediated innate host defenses that it can survive and concomitantly suppresses Th1/Th2-driven adaptive immune responses that would eliminate it. Here we show that intravaginally administered IL-12 encapsulated in sustained-release polymer microspheres led to significantly faster clearance of infection. Enhanced resistance to secondary infection with the same strain of N. gonorrhoeae induced by intravaginal treatment with IL-12 microspheres during primary infection persisted for 6 months. Furthermore, treatment of mice with IL-12 microspheres during gonococcal challenge induced resistance to reinfection with unrelated strains of N. gonorrhoeae. ELISA, flow cytometry and PCR studies demonstrated that IL-12 microsphere treatment in infected mice effectively enhanced Th1 immune responses in the vaginal tract and the draining lymph nodes, without hindering Th17 responses. Although IL-12 microspheres did not change Th2 responses, the treatment significantly increased the production of gonococcus-specific circulating IgG and vaginal IgA and IgG antibodies. These results suggest that local administration of microencapsulated IL-12 may serve as a novel, effective immunomodulatory and therapeutic strategy against gonorrhea.
Pneumococcal choline-binding proteins contain an activity that mediates Streptococcus pneumoniae-induced immunosuppression (IRC8P.478)
Authors: Saumyaa Saumyaa , Colino Jesus , Snapper Clifford 
Published: January 2023

Abstract
We previously demonstrated that intact, inactivated Streptococcus pneumoniae (strain R36A) inhibits murine IgG, T follicular helper, germinal center and plasma cell responses to co-immunized chicken ovalbumin (cOVA), whereas intact, inactivated Streptococcus agalactiae and Neisseria meningitides did not. Since phosphorylcholine (PC), reported to mediate immunosuppression by filarial organisms, was selectively expressed by R36A, we determined whether R36A lacking PC (R36APC-) lost this inhibitory activity. R36APC- indeed, exhibited a significant loss of inhibition of the anti-cOVA response. However, R36APC- is also devoid of a family of choline binding proteins (CBPs) that are attached to the bacterial cell surface via PC binding. We thus prepared CBP-depleted R36A (R36ACBP-) by stripping CBPs from the cell surface using choline chloride. R36ACBP-, which continues to express PC, also exhibited a significant reversal in inhibition of the anti-cOVA response. Inhibition of the anti-cOVA response was also mediated by isolated R36A cell walls, supernatant from choline chloride-treated R36A, and purified CBPs. In contrast, trypsinized intact R36A and trypsinized CBP-containing supernatant lost their inhibitory activity. These results suggest a novel immunosuppressive property expressed by CBPs and CBP-expressing S. pneumoniae. Studies are continuing to specifically identify this activity.
Bias in synapse stability leads to suppression of naive CD8 T cell activation by memory CD8 T cells during secondary responses (IRC9P.709)
Authors: Mayya Viveka , Judokusumo Edward , Medina Ricardo , Zairis Sakellarios , Wiggins Chris , Kam Lance , Dustin Michael 
Published: January 2023

Abstract
T cells can form either a stable synapse or a mobile junction termed kinapse when interacting with antigen presenting cells during an immune response. Stable synapses can lead to long dwell times with antigen presenting cells, whereas mobile kinapses likely favor transient interactions with antigen presenting cells. The underlying cell-intrinsic mechanisms determining the mode of interaction as well as their specific immunological consequences are not known. We have implemented methods borrowed from computer-vision and machine-learning disciplines to accurately quantify the positional stability of T cell junctions formed on model antigen presenting surfaces coated with ICAM-1 and anti-CD3. We have found that human memory CD8 T cells form stable synapses whereas naive CD8 T cells form kinapses or transient contacts. This bias in synapse stability is neither observed in human CD4 T cell subsets nor in murine CD8 T cell subsets under the conditions we have tested. Increased surface expression of LFA1 in human memory CD8 T cells contributes to their increased synapse stability. Using patterned surfaces to simulate competition for antigen presenting cells, we show that synapse stability exhibited by memory CD8 T cells causes suppression of naive cell activation by preventing access to antigen. Increased synapse stability of memory CD8 T cells is expected to promote the development of CTL escape variants during hyper-mutable viral infections and thus prove detrimental to the host.
Helper T cell differentiation and regulation of inflammatory lymphangiogenesis (IRC10P.468)
Authors: Elhadad Sonia , Mehrara Babak 
Published: January 2023

Abstract
Lymph node lymphangiogenesis (LNL) is a critical regulator of inflammation. Although it is clear that pro lymphangiogenic factors VEGF-A and C play a central role in the regulation of this process, recent studies showed that these effects are balanced with anti-lymphangiogenic forces like IFN-g. However, although it is clear that T cells play a negative regulatory role in LNL, the effect of T cell differentiation remains unknown. We analyzed the effects of T cell differentiation on LNL in a mouse model of inflammation and analyzed draining lymph nodes at various time point by flow cytometry, histology, and RT-PCR. We found that induced inflammation, resulted in a robust inflammatory response in the draining lymph nodes beginning 7 days after injection and lasting until day 21. This process was associated with generation of a mixed Th1,Th2 responses, decreased expression of IFN-g, and increased expression of VEGF-A/C. Loss of CD4 or CD8 T cells resulted in augmentation of lymphangiogenesis, increased expression of IFN-g, and VEGF-C. Treatment of mice with IFN-g, IL13, or IL17A neutralizing antibodies resulted in augmented LNL and increased expression of VEGF-C and IFN-g. Loss of either CD4 or CD8 T cells resulted in augmentation of LNL as a result of increased VEGF-A/C expression. Similarly, inhibition of Th1, Th2, or Th17 cytokines increase LNL and increased VEGF-A/C expression. In conclusion, we showed that T cell differentiation modulates inflammatory LNL.
Evaluation of IL-2 therapy on human immune system homeostasis in humanized mice (LYM8P.731)
Authors: Durost Philip , Shultz Leonard , Brehm Michael 
Published: January 2023

Abstract
Regulatory T cells (Treg) are an IL-2 dependent subset of CD4 T cells with the capacity to suppress immune responses. In mice, IL-2 treatment can prevent autoimmune disease and rejection of allogeneic tissues by selective expansion of Treg populations. Moreover clinical trials have shown that low dose IL-2 therapy increases Treg in humans and is effective in ameliorating immune disorders. However, low dose IL-2/rapamycin therapy in type 1 diabetic individuals was less effective and had potentially detrimental effects on beta cell function. The unpredictability of IL-2 therapy in humans emphasizes the need for novel models to fully evaluate therapies targeting human Treg before being used in the clinic. Here we have evaluated the impact of IL-2 treatment on human immune systems using a humanized mouse model. For these studies we used NOD-scid IL2rγnull (NSG) mice engrafted with fetal thymus and liver tissues and human HSC [NSG-BLT (bone marrow, liver, thymus) mice]. Human T cell populations develop efficiently in NSG-BLT mice making this model ideal to study T cell specific therapies. NSG-BLT mice treated with an AAV vector expressing human IL-2 showed a significant increase in the number of both functional human Tregs and human NK cells. IL-2 expression did not alter T effector/memory T cells subsets. Collectively, these data show that humanized mouse models are an ideal platform for testing human-specific immune therapies.
The role of CD4+ T cell subsets in the clearance of Pneumocystis murina pneumonia in mice (MPF6P.734)
Authors: Zheng Mingquan , Ripple Michael , Yu Amy , Cleveland Hillary , Stefanovski Matthew , Chen Kong , Kolls Jay 
Published: January 2023

Abstract
Pneumocystis (PC) Pneumonia remains a serious complication in the immuno-compromised host. The host defense against PC is critically dependent upon CD4+ T cells. The role of specific T-cell subsets that mediate host resistance against PC infection remains unclear. We use STAT transcription factor knockout (KO) mice to determine the role of Th1, Th2, and Th17 cells in PC infection. After 28 days infection with PC, serum, lung tissue, and mediastinal lymph node (LN) were collected. LN and lung tissue cells were measured for cytokines by ELISpot and intracellular staining. Total lung RNA was measured for PC burden by RT-qPCR. Cytokines were assayed from cell culture supernatant. Anti-PC antibody was measured by ELISA.Stat4, Stat6 single KO, and Stat4/6 double KO mice all were resistant to PC compared to the Rag2/gc KO mice. Stat4 was required for a Th1 response and Stat6 was required for a Th2 response. STAT3fl/fl/CD4Cre+ mice had partial susceptibility of PC. Stat3 expression was required in CD4+ T cells for PC specific Th17 responses. STAT4/6 double KO crossed to STAT3fl/fl/CD4Cre+ (Triple KO) mice were highly susceptible to PC infection compare to single and double KO mice. Individual Stat4 and 6 and double mutants clear PC. STAT3fl/fl/CD4Cre+ conferred partial susceptibility to PC. Triple KO were as susceptible for PC as Rag2/gc KO mice. These data support the hypothesis that PC is a true opportunist and that various T-cell subsets can confer protection against infection.
Cell stress molecules expression in periodontal disease (MUC7P.762)
Authors: Aguilera Galaviz Luis , Aceves Medina Maria , Hernandez Montoya Martha , Diaz Rosas Cristal , Frausto Esparza Silverio , Estrada Garcia Iris 
Published: January 2023

Abstract
Periodontal Disease is bacterial infection of tooth-supporting tissues leading to chronic inflammation and loss of teeth, the immunopathologic mechanisms include inflammatory cells and proinflammatory cytokines; persistent local inflammation develops the expression of stress molecules as Hsp90, Hsp70 and P53 also autoantibodies. The rol of cellular stress and autoimmunity in periodontal disease is not understood. Objetives: Determine the expression of cell stress molecules and auto-antibodies in periodontal disease patients. Methods: 34 patient with severe chronic periodontitis and 14 healthy people were clinical evaluated. Periodontal tissue and sera were obtained. Electroforesis, Western blot and Immunodetection to identify Hsp70, Hsp90 and P53 was set up, Hsp 90 were evaluated by RT-PCR and IFI for ANCA, antiepithelial antibodies in mouse skin and ANA using Hep2 cells. Results: 57% were positives for ANCA, total of sera were positives for nuclear and/or cytoplasmic antibodies and 57% presented anti-epithelial antibodies, Hsp90, Hsp70 and P53 is expressed according with the severity of disease. Conclusions: The expression profile of Hsps in periodontal tissue is according with the severity, and could be associated with cellular stress because of the damage mechanism induced by periodontal pathogens and trigger necrotic or apoptotic phenomena. Destruction of extracellular matrix in periodontal tissue could be mediated by auto-antibodies that recognize self structures.
Human chorionic gonadotropin supports human pregnancy by induction of regulatory T cells (MUC7P.766)
Authors: Schumacher Anne , Poloski Eileen , Zenclussen Ana 
Published: January 2023

Abstract
Regulatory T cells (Treg) are proposed to be involved in fetal tolerance induction. However, the precise mechanisms underlying their generation and function are still not defined. Recently, we proved that the pregnancy hormone human Chorionic Gonadotropin (hCG) efficiently attracts human Treg to trophoblasts favoring a local Treg accumulation. Here we analyzed whether hCG is also involved in Treg generation and activity. Naïve T cells were isolated from blood of pregnant women and co-cultured with either hCG-producing trophoblasts (JEG-3, primary cells) or non-hCG-producing trophoblasts (HTR8, Swan71) and keratinocytes (HaCat). To confirm hCG specificity, we either blocked hCG in JEG-3 co-cultures or used urine-purified or recombinant hCG as culture supplements. Treg induction was then assessed by the expression of specific Treg markers. Furthermore suppressive function of hCG-treated naïve T cells was evaluated in a mixed leukocyte reaction. Co-culture of naïve T cells with JEG-3, primary trophoblasts or supplemented hCG resulted in a statistically significant increase in the expression of specific Treg markers as compared to co-cultures with HTR8, Swan71 or HaCat cells. Moreover, blockage of hCG in JEG-3 co-cultures impaired the up-regulation of Treg markers. Interestingly, we further confirmed an increased suppressive activity of naïve T cells after hCG treatment. Our results strongly suggest a key role for hCG in Treg generation and induction of suppressive activity.