bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interleukin-4 directly promotes intestinal mast cells expansion in the context of food allergy (P3256)
Authors: Burton Oliver , Darling Alanna , Zhou Joseph , Noval-Rivas Magali , Jones Tatiana , Gurish Michael , Chatila Talal , Oettgen Hans 
Published: January 2023

Abstract
Interleukin-4 is a pivotal cytokine in the development of allergic disorders. Mast cells are key effectors of food allergen-driven anaphylaxis, and IL-4 has been implicated in their function. More recently, however, we and others have shown that IL-4 has a mix of inhibitory and stimulatory effects on bone marrow-derived mast cells. We therefore sought to clarify the direct effects of IL-4 on intestinal mast cell function and growth. In this study, we demonstrate that IL-4 drives a marked intestinal mastocytosis in mice. Similar effects were observed in animals expressing Il4rαF709, an activating variant of the IL-4 receptor α-chain that confers susceptibility to food allergen-driven anaphylaxis. Il4rαF709 mice show heightened sensitivity to IgE-mediated anaphylaxis without increased sensitivity to mast cell mediators. Exposure to IL-4 upregulates anti-apoptotic Bcl proteins in intestinal mast cells, leading to enhanced survival and proliferation via STAT6. These effects are intensified by the Il4rαF709 signaling variant. Mixed bone marrow chimera experiments demonstrate that Il4rαF709 mast cells hold a substantial competitive advantage over wild-type cells in populating the intestine in response to food allergen stimulation. IL-4Rα-deficient mast cells do not effectively expand, establishing a cell-intrinsic effect of IL-4 in intestinal mast cell homeostasis. Our results demonstrate that IL-4-signaling is a key determinant of mast cell expansion in food allergy.
Use of in vivo caspase inhibitor in rat burn and infection injury prevents gut bacterial translocation and modulates CD4+ T cell effector immune response. (P3272)
Authors: Fazal Nadeem 
Published: January 2023

Abstract
Our previous studies have focused on gut-associated lymphoid tissues, failure of intestinal gut barrier, bacterial translocation, and sepsis exploring the roles of antigen-presenting cells, DCs, CD4+ T cell and regulatory T cells in immunosuppression following acute burn and/or burn-plus-infection injury. In an established rodent model of burn plus infection we tested in vivo use of caspase inhibitor and studied T cell effector immune response and gut barrier function. Z-VAD-FMK, a highly specific, cell-permeable, and irreversible inhibitor of caspases was given to rats and were then assessed for mortality, bacterial translocation, T cell receptor expression and apoptosis. The results suggested an increase in CD4+CD25+ T cell phenotype in splenocytes of burn as compared to sham rats, however such an elevation in CD4+CD25+ expression was reduced in CLP and CLP+Burn+Caspase-Inhibitor groups. The results also showed that there was an increase in adhesion receptors (LFA) and decrease in CD62L in burn, whereas no difference in these adhesion receptor expressions was ascertained in Burn+CLP and Burn+CLP+Caspase-Inhibitor treated rats. A decrease in homing receptor (CD49d) was also noticed in Burn+CLP and Burn+CLP+Caspase-Inhibitor-treated animals. The Colony Forming Unit (CFU) results showed absence of bacterial translocation in rats that were subjected to burn+CLP+Caspase-Inhibitor, however there was significant bacterial translocation in rats from the group of CLP and burn only.
Allergen-induced lysosomal CCL20 release from bronchial epithelial cells (P3284)
Authors: Lajoie Stephane , Webb Mark , Wills-Karp Marsha 
Published: January 2023

Abstract
Although allergic asthma is a chronic inflammatory disease of the lung mediated by Th2 cells, the pathways leading to this response are not clear. Recent studies have highlighted influence of pulmonary dendritic cells (DCs) on T-cell differentiation. Rapid DC recruitment into the bronchial mucosa has been well documented in animals and humans in response to allergen. Central to early DC recruitment, CCL20 is the only known ligand for CCR6, a receptor preferentially expressed on immature DCs. The potential importance of CCL20/CCR6 in allergic sensitization was demonstrated in a cockroach model of allergic asthma in which Ccr6-/- mice were protected against asthma. At present, the mechanisms by which allergen induces secretion of DC-attractant chemokines are unknown. We previously showed that bronchial epithelial cells release CCL20 relatively rapidly after house dust mite (HDM) (Nathan, 2009). This induction was not prevented by the inhibiting protein synthesis (cycloheximide) suggesting release through a transcriptionally independent mechanism. These data suggested that HDM may induced the release of preformed stores of CCL20
Glucocorticoid-induced TNF receptor family related protein ligand regulates the egress of monocytes from the spleen (P3288)
Authors: Liao Gongxian , van Driel Boaz , Magelky Erica , O’Keeffe Michael , Herzog Roland , Mizoguchi Emiko , Bhan Atul , Terhorst Cox 
Published: January 2023

Abstract
Glucocorticoid-Induced TNF Receptor family-related protein Ligand (GITR-L) is expressed on antigen presenting cells. Surprisingly, GITR-L−/−Rag−/− mice develop a markedly milder colitis than Rag−/− mice upon transferring CD4+CD45RBhi T cells or by administering agonistic anti-CD40 antibody. In both colitis models as well as in a peritonitis model a reduced number of Ly6C+CD11b+MHCII+ macrophages was found in the inflamed sites of GITR-L−/− mice. By contrast, the number of non-differentiated monocytes was approximately three-fold higher in the spleen of inflamed GITR-L−/−Rag−/− mice than in Rag−/− mice. The formation of an active dimer of the angiotensin II type 1 receptor (AT-1) by splenic monocytes, which regulates their egress, was dynamically affected by the GITR-L deficiency. Infusion of Angiotensin II caused retention of splenic monocytes in GITR-L−/− but not wt mice. We conclude that upon induction of inflammation GITR-L regulates egress of monocytes from the spleen, thus impacting intestinal homeostasis in mice.
Resveratrol and its derivatives modulates TH17 differentiation and attenuates experimental autoimmune encephalitis (P5139)
Authors: Yen Hung-Rong , Chung Pei-Chen , Yang Hsu-Ling 
Published: January 2023

Abstract
The pathogenic role of IL-17 and TH17 cells in inflammation and autoimmunity has prompted the development of new inhibitors by pharmacologic approaches. Here we demonstrated resveratrol (3,5,4'-trihydroxy-trans-stilbene) and its derivatives from natural products and traditional Chinese medicine, inhibit TH17 differentiation in a concentration-dependent manner in vitro. CD4 T cells polarized in the TH17 condition with resveratrol expressed a relatively lower level of IL-17, ROR-gt, and TH17-related gene expression compared to the non-treated cells. To further determine its efficacy in vivo, we administered candidate drugs in a murine model of experimental autoimmune encephalitis, resveratrol and its derivatives ameliorates the disease severity and the production of TH17 in vivo. Our preliminary data fit the niche that is required to modulate TH17 for immunotherapy and we suggest that, to treat autoimmune disease, administering resveratrol and its derivative could be a promising pharmacologic approach to achieve the goal. Additional mechanisms are being explored.
Resveratrol ameliorates lupus by epigenetic modulation of Foxp3 and IL-17 genes resulting in differential regulation of regulatory T cells and Th17 cells via activation of aryl hydrocarbon receptor (P5129)
Authors: Singh Narendra , Singh Udai , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Resveratrol (RES), a polyphenolic compound found in plant products, possesses anti-inflammatory, antioxidant, and anticancer properties. We investigated the use of RES for the treatment of lupus using an animal model. RES treatment significantly suppressed Pristane-induced lupus in SJL/J mice. There was significant downregulation of IL-17 but upregulation of FoxP3 expression. RES initiates its early signaling via AhR and recent studies have suggested that AhR may regulate T cell differentiation. We examined the effect of RES on differentiation of Tregs and Th17 cells, microRNA (miR) profile in T cells, and epigenetic modulation of Foxp3 and IL-17 genes. There was a significant difference in the miR profiles in T cells of RES-treated mice, when compared to vehicle-treated mice. Analysis of 609 miRs revealed dysregulation of 112 miRs including downregulation of miRs (miR-31,-490, -217, and -190) that regulate FoxP3 and upregulation of miRs (miR-200, -203, -431, -494, and -495) that regulate IL-17. Upon examination of epigenetic effects of RES on Foxp3 and IL-17 promoters, we observed significant demethylation of Foxp3 promoter but methylation of IL-17 promoter. Culture of T cells in the presence of RES led to significant increase in Tregs but decrease in Th17 cells in vitro and in vivo. Together, these data demonstrated that RES via AhR regulates epigenetic modulation and can have differential effects on T cell differentiation into Tregs/Th17 cells causing suppression of lupus.
Critical role of all-trans retinoic acid in stabilizing human nTregs under inflammatory conditions (P5133)
Authors: Zheng Song Guo , Wang Julie , Horwitz David , Quismorio Francisco , Lu Ling 
Published: January 2023

Abstract
Recent studies have demonstrated that thymus-derived naturally-occurring CD4+Foxp3+ regulatory T cells (nTregs) in both human and mouse are unstable and dysfunctional in the presence of pro-inflammatory cytokines. All-trans Retinoic Acid (atRA), the active derivative of vitamin A, has been shown to regulate Treg and T effector cell differentiation. Herein, we report that atRA prevents human nTregs from converting to Th1 and/or Th17 cells and sustains their Foxp3 expression and suppressive function in vitro or in vivo following encountering with IL-1 and IL-6. Interestingly, adoptive transfer of human nTregs pre-treated with atRA significantly enhanced their suppressive effect on xeno-graft versus host diseases (xGVHD) and only these cells after stimulating with IL-1/IL-6 sustained the functional activity against xGVHD in a humanized animal model. atRA suppresses IL-1R upregulation and accelerates IL-6R downregulation and diminishes their signaling events, as well as increases histone acetylation on FOXP3 gene promoter and CpG demethylation in CNS regions of FOXP3 gene locus. These results strongly implicate that nTregs primed with atRA may represent a novel treatment strategy to control established chronic immune-mediated inflammatory diseases.
Investigating TLR4-signaling mechanisms in CHME-5 human microglial cells and the effects of β-funaltrexamine treatment (P1236)
Authors: Figueroa Leandra , Das Subhas , Buck Daniel , Stevens Craig , Davis Randall 
Published: January 2023

Abstract
Neuroinflammation is instrumental in neuronal damage observed in various neuropathologies. Thus, understanding pro-inflammatory pathways of human glial cells is important in identifying potential pharmacological strategies to modulate inflammatory signaling in these cells. Microglia-derived cytokines and chemokines are instrumental in neuroinflammation and several proinflammatory mediators activate these cells to include bacterial lipopolysaccharide (LPS) that signals through toll-like receptor 4 (TLR4). Microglial TLR4 signaling is not well studied; therefore, our goal is to investigate the molecular mechanisms of TLR4 microglial signaling, which entailed treatment of human microglial cells (CHME-5) with LPS. LPS induced NF-κB activation, demonstrated by increased NF-κB p65 binding activity and phosphorylated p65 expression in CHME-5 cells. TLR4 expression also increased alongside NF-κB activation. On the other hand, co-exposure to the μ-opioid receptor antagonist, β-funaltrexamine (β-FNA), down-regulated LPS-induced NF-κB activation. To date, our data suggest that the inflammatory actions of microglial TLR4 signaling may be partly due to modulation of NF-κB activation. Thus, we have also initiated studies to elucidate the effects of LPS and β-FNA on other key TLR4 signal transduction proteins. Together, these ongoing studies are expected to further elucidate the mechanisms of LPS-induced TLR4 signaling in human microglia.
Herpes simplex virus type 2 induces innate immune response via TLR4-MyD88/Mal-NF-kB axis in human cervical epithelial cells (P1220)
Authors: Li Hui , Liu Hongya , Chen Kai , Feng Wenqiang , Wu Xinxing , Liu Xuefeng 
Published: January 2023

Abstract
Toll-like receptors (TLRs) play a crucial role in regulating innate immune response to HSV-2 infection. Current studies on TLRs and viral infection have used either immuno-competent cells and/or murine systems. The innate immune pathway in response to this virus in its natural host cells remains unclear. We have established an in vitro HSV-2 acute infection model with Human cervical epithelial (HCE cells, the primary target and natural host cells for HSV-2) to investigate the role of TLRs-mediated innate immune response to HSV-2. In current study, we found that HSV-2 infection induced NF-kB reporter activity and expression of cytokines are TLR4-dependent. Knockdown experiments demonstrated that the adaptor molecules MyD88 and Mal of the TLRs signaling pathways are required in the HSV-2 induced TLR4-dependent NF-kB activation in HCE cells. Western blot assay suggested that knockdown of TLR4 decreased the phosphorylation of IRAK1 and inhibitor of NF-kB (IkB-a) upon HSV-2 infection. Finally, decreased expression of either TLR4 or MyD88/Mal alone or both significantly abolished productions of IL-6 and IFN-beta by ELISA analysis. Taken together, our results reveal for the first time that there exists the pathway via TLR4-Mal/MyD88-IRAK1-NF-kB axis in response to HSV-2 infection in its natural host cells.
MeCP2 enforces Foxp3 expression to determine natural regulatory T cells’ resilience to inflammation (P1023)
Authors: Li Chaoran , Jiang Shan , Sevilla Jose , Li Qi-Jing 
Published: January 2023

Abstract
Foxp3+ regulatory T cells are crucial for peripheral tolerance and immune homeostasis. During inflammation, the steady expression of Foxp3 in mature regulatory T cells is essential for maintenance of their lineage identity and suppressive function. However, the molecular machinery governing regulatory T cells’ resilience to inflammation induced Foxp3 destabilization is not fully understood. Here, we demonstrate that Methyl-CpG binding protein 2, an eminent epigenetic regulator known primarily as the etiological factor of the neurological disorder Rett syndrome, is essential to sustain regulatory T cells’ Foxp3 expression during inflammation. In response to inflammatory stimuli, Methyl-CpG binding protein 2 is specifically recruited to the CNS2 region of the foxp3 locus, where it collaborates with CAMP responsive element binding protein 1 to promote local histone H3 acetylation, thereby counteracting inflammation induced epigenetic silencing of foxp3. Consequently, regulatory T cell specific deletion of Methyl-CpG binding protein 2 is sufficient to induce spontaneous immune activation in young adult mice. Furthermore, we demonstrate that Foxp3 expression diminishes with time in Methyl-CpG binding protein 2 deficient regulatory T cells, resulting in their failure to suppress Th17-mediated colitis. Thus, Methyl-CpG binding protein 2 serves as a critical safeguard that confers regulatory T cells with resilience against inflammation.
The induction of anterior chamber associated immune deviation with Type II collagen (P1045)
Authors: Ashour Hossam , Farooq Shukkur 
Published: January 2023

Abstract
The administration of antigens into the anterior chamber (AC) of the eye induces a special form of antigen-specific peripheral immune tolerance termed AC-associated immune deviation (ACAID), which prevents delayed-type hypersensitivity (DTH) responses and other inflammatory responses. Type-II collagen (CII) is highly expressed in cartilage tissues and has been linked to Rheumatoid arthritis, aging, and osteoarthritis. To explore the potential for ACAID induction via CII, we checked for different signs of ACAID generation following the AC injection of CII in BALB/c mice. We hypothesized that the mechanism of ACAID induction involves efferent T regulatory cells (Tregs). Both local adoptive transfer (LAT) assays and DTH assays were performed. Results indicated that ACAID induction was driven by the AC injection of CII. Spleen cells of mice injected with CII in the AC significantly suppressed DTH responses. ACAID induction was mediated by efferent Tregs in the spleen. CII-mediated ACAID induction could have multiple therapeutic implications, especially in autoimmunity.
Establishing the molecular size threshold of immune synapse penetration: Implications for drug access (P4263)
Authors: Cartwright Adam , Griggs Jeremy , Davis Daniel 
Published: January 2023

Abstract
Natural Killer (NK) cells survey target cells through activating, inhibitory and adhesion receptors that bind their ligands at the immune synapse. This dense mesh of protein-protein interactions at the intercellular contact may create a seal, or gasket, around the synapse to prevent the leakage of secreted proteins, but this remains poorly understood. Nanometer scale fluorescent dextrans of known sizes were used to determine the size threshold of molecules that could penetrate the immunological synapse between NK and target cells, by measuring fluorescence accumulation within the intercellular region. Dextran was completely excluded from the synapse when its hydrodynamic diameter was above 13nm, whereas a 4nm dextran was able to diffuse in and out of the synapse. This showed that the synapse represents a physical gasket through which molecules can penetrate in a size-dependent manner. This establishes a size threshold of molecules that are able to enter the synapse. Drugs which target proteins within the synapse should be engineered to these dimensions in order to penetrate the synapse.
Topical CpG adjuvant enhances immune response to subcutaneous antigen by modulating the cutaneous lymph node environment (P4267)
Authors: Cheng Wing-Ki (Vicki) , Plumb Adam , Wee Kathleen , Kollmann Tobias , Abraham Ninan , Dutz Jan 
Published: January 2023

Abstract
Current vaccines are mainly administered by intramuscular route but the skin harbors many immune cells and can be used as a site of immunization. Topical CpG oligodeoxynucleotide (ODN), a Toll-like receptor 9 (TLR9) agonist, improves humoral and cell-mediated immune responses to locally injected protein-based vaccines in mice but the mechanisms are still unclear. Topical CpG ODN is internalized by CD11c+ cells that are detected within the skin draining lymph nodes (SLN) at 48 hours post treatment. Ablation of TLR9 in the hematopoietic compartment of bone marrow chimeric mice abrogates antigen-specific CD8+ T cells production. TLR9 ablation in the stromal compartment also decreases topical CpG adjuvant effect. Topical CpG ODN differentially modulates the environment of SLN compared to subcutaneous administration within 24 hours of treatment (with 4-7 fold increased expression of 4 genes: Ccl4, Cxcl3, Ifng and Il11). A higher proportion of antigen-specific CD4+ T cells in the SLN express tissue-homing molecules (P- and E-selectin ligand) when adjuvant is administered topically compared to subcutaneously. Egress of lymphocytes from SLN is necessary for optimal CpG adjuvant effect using a contact hypersensitivity model. We propose that topical CpG ODN activates stromal cells as well as hematopoietic-derived cells in the skin, to subsequently modulate the environment of SLN. These changes enhance protective humoral and cell-mediated immune responses.
B7-H1 blockade plus regulatory T cell depletion improves clinical efficacy over either strategy alone in a melanoma model (P4268)
Authors: Curiel Tyler , Murthy Kruthi , Kious Mark , Sun Lishi , Zhang Bin , Pandaswara Srilakshmi , Hurez Vincent 
Published: January 2023

Abstract
Denileukin diftitox (DD) is an FDA-approved conjugate of human IL-2 + diphtheria toxin. It depletes Tregs and improves clinical and immune outcomes, but Treg generation can reduce efficacy. B7-H1 is an immune co-signaling molecule that promotes Treg generation and hinders anti-cancer immunity. αB7-H1 or DD each slowed tumor growth and improved anti-tumor immunity in B16 melanoma. DD reduced Treg numbers but αB7-H1 also reduced Treg suppression. In vitro, αB7-H1 reduced Treg generation from naïve CD4+CD25- T cells. We thus hypothesized that αB7-H1 would synergistically improve clinical efficacy of Treg depletion with DD by slowing regeneration or function of Tregs after depletion. We treated mice 7 days after B16 challenge when detectable tumors were present. DD+αB7-H1 significantly slower tumor growth at 25 days (avg tumor ~600 mm3 vs either single agent treatments (avg 1500-2000 mm3; p &lt;0.05). Spleen CD4+ and CD8+ T cells with combined therapy showed significantly more activated CD44+CD62L- cells vs individual treatments, but we found no significant difference in T cell interferon-γ or IL-17 in treatment groups. Strikingly, we also did not detect reduced Treg function or numbers in DD + αB7-H1 combination vs individual treatments in spleen cells. Preliminary data suggest that effects are primarily in tumor draining lymph nodes, which are under current study. We use FDA approved agents like DD or agents in clinical trials such as αB7-H1 for rapid clinical translation.
Adiponectin deficiency suppress murine fibrosarcoma growth via modulation of tumor-associated macrophages polarization (P2127)
Authors: Peng Jiao , Ho Derek , Chan Vera , Tam Paul , Chen Yan 
Published: January 2023

Abstract
Adiponectin was reported to modulate macrophage polarization towards an anti-inflammatory phenotype (M2-type), but it remains unclear whether APN deficiency could modulate tumor development via tumor-associated macrophages. In this study, effects of APN on TAMs polarization and tumor development were investigated in MN/MCA1 inoculated mouse fibrosarcoma model. Our results indicated that APN deficiency exerted suppressive activities on the development of MN/MCA1 fibrosarcoma in vivo, whereas no pronounced APN-induced cyto-toxicity was observed in vitro. Phenotypic analysis demonstrated that M1-like TAMs were accumulated in tumor bearing APN-/- mice, which is evidenced by expression of M1-like marker, MHCII molecule. Moreover, M1-like markers, such as iNOS, TNF-α and IL-12 were up-regulated and the M2-like makers IL10, YM1, IL4Rα, MgI-1and MMP-9 mRNA were down-regulated in TAMs isolated from APN-deficient mice. Immunostimulatory effects were also depicted by increasing tumor infiltrated cytotoxic T lymphocytes and natural killer cells. In summary, adiponectin deficiency could promote TAMs polarization towards M1-like phenotype which might provide insight of modulation of TAMs in cancer therapy.