bluuebunny/crossref_metadata_2025_split huggingface dataset:
DC-specific HMGB1 KO mice clear vaccinia viral infection more effectively (VIR5P.1018)
Authors: Wang Wenqian , Vernon Philip , LI Guanqiao , Billiar Timothy , Sampath Padmavathi , Stephen Thorne , Morelli Adrian , Liang Xiaoyan , Lotze Michael 
Published: January 2023

Abstract
High-mobility group box 1 (HMGB1), an evolutionarily ancient and abundant DNA-binding protein within the nucleus, acts as a Damage Associated Molecular Pattern (DAMP) molecule extracellularly to promote immunity. It also functions in the cytosol to promote autophagy, a critical mechanism in viral clearance. HMGB1 is required for dendritic cell (DC) maturation and chemotaxis but how HMGB1 regulates DC function particularly in viral clearance is unclear. We constructed mice with HMGB1-knockout (KO) DCs (DCH) by crossing HMGB1 floxed mice with CD11c Cre mice. Although KO DCs differentiate normally from the bone marrow, they have diminished lymphocyte stimulatory capacity and polarize naive T cells towards Th17 and Treg phenotypes while wide type (WT) DCs induce IFNγ and IL-4 secretion at a ratio of 1:1 in allogeneic assays (p&lt;.05). We infected DCH and WT mice with luc+ vaccinia and assayed viral infection over 9 days. Surprisingly, the virus is cleared more rapidly in the DCH cohort (p&lt;.05). This is associated with increased T cells (p&lt;.01) and CD11c+ cells (p&lt;.05) within the spleen. We hypothesize that the proliferation of vaccinia depends on intact autophagy which may be impaired in KO DCs. We also find DCs release HMGB1 through exosomes, which are membrane vesicles enriched in MHC molecules and responsible for virus transmission. These results suggest that HMGB1 is not only critical for function but also contributes to the persistance of vaccinia virus in this murine model.
Increased pathology in herpes stromal keratitis results from differences in local, and not systemic, immune responses (VIR5P.1028)
Authors: Sullivan Brooke , Rowe Alexander , Hendricks Robert 
Published: January 2023

Abstract
Harboring latent herpes simplex virus type 1 (HSV-1) in the trigeminal ganglion (TG) and periodic shedding of virus at the corneal surface can be asymptomatic or can trigger an immunopathological response called herpes stromal keratitis that leads to progressive scarring and visual impairment. Our goal is to understand the events that differentiate an asymptomatic from a pathological HSV-1 infection. To this end we infected mice with two different strains of HSV-1: RE that causes progressive corneal opacity, and KOS that induces no overt corneal pathology. The kinetics of viral clearance and the latent viral load in the TG at 30 days post-infection (dpi) were similar following corneal infection with the two HSV-1 strains. However, the pathogenic RE strain induced a stronger CD4 and CD8 T cell response in the draining lymph nodes and TG at 8 dpi that persisted in the TG through 30 dpi. Interestingly, bilateral infections with RE in one eye and KOS in the other resulted in pathology only in the RE infected eye, suggesting that the increased immunopathology caused by the RE strain reflects local differences in the cornea or ipsilateral TG, and not differences in the systemic immune response. We are currently testing the hypothesis that greater viral gene expression by the RE strain during latency results in a larger T cell infiltrate in both the cornea and the TG.
Regulation of lupus-antigen specific B cells by follicular dendritic cells (BA12P.113)
Authors: Das Abhishek , Dwivedi Nishant , Chatterjee Priyadarshini , Carroll Michael 
Published: January 2023

Abstract
Humoral responses are central to many autoimmune diseases such as systemic lupus erythematosus (SLE). Yet little is known about the cells and factors that are vital for maturation and sustenance of autoreactive B cells. In this study, we used a B cell receptor (specific for RNP complexes) knock-in strain 564Igi to demonstrate an important role of follicular dendritic cells (FDCs) in loss of tolerance of autoreactive B cells. Breeding the lupus-prone strain to a C4-deficient background resulted in escape of tolerance of auto-reactive idiotype+ (Id+) B cells. Total ablation or in vivo de-differentiation of FDCs in the C4-/- 564Igi mice resulted in a dramatic decrease in auto-reactive B cells and germinal centers suggesting their role in promoting loss of tolerance. Gene expression analysis of FDCs isolated from C4 -/- 564Igi mice show an increased expression of IFN-alpha (IFN-a) and downstream response genes. Treatment of the mice with anti-IFN-a receptor (IFNAR) resulted in restoration of tolerance. Blockade of myeloid cell IFNAR was identified as the critical pathway as mixed marrow chimeras of C4-/- 564Igi and IFNAR- deficient bone marrow into C4-/- recipients have a significantly lower frequency of auto-reactive B cells, which underlines the importance of IFN-a signaling in myeloid cells. Hence, we propose a model in which IFN-a secreted by activated FDCs act on dendritic cells, which in turn supports auto-reactive B cells by producing factors like IFN-a and BAFF.
Elucidating the role of TNF-like weak inducer of apoptosis (TWEAK) in the pathogenesis of cutaneous lupus (BA12P.114)
Authors: Doerner Jessica , Wen Jing , Xia Yumin , Friedman Adam , Michaelson Jennifer , Burkly Linda , Wu Lan , Putterman Chaim 
Published: January 2023

Abstract
Cutaneous manifestations are very common in the systemic autoimmune disease systemic lupus erythematosus (SLE). TWEAK is a soluble cytokine member of the TNF superfamily that binds to a sole receptor, Fn14. TWEAK/Fn14 signaling is involved in cell survival, apoptosis, cytokine production, and angiogenesis, and as such has been found to be important in both tissue repair and inflammatory diseases. Whether TWEAK is involved in autoimmune skin disease is not known. We found that lupus-prone MRL/lpr mice deficient for Fn14 (Fn14KO) had attenuated cutaneous disease as compared to Fn14WT mice. MRL/lpr Fn14KO mice had significantly less epidermal acanthosis and follicular plugging, and there was decreased infiltration of T cells and macrophages as compared to Fn14WT mice. In vitro, TWEAK treatment of murine keratinocytes stimulated the secretion of RANTES (via Fn14), and promoted apoptosis. Parthenolide decreased production of RANTES, indicating that this effect of TWEAK is mediated via NF-KB. Ultraviolet light, specifically UVB, is recognized as a potent trigger of cutaneous lupus. We found that TWEAK co-treatment exacerbates UVB light- induced keratinocyte apoptosis and increases the amount of RANTES secreted, when compared to cells treated with UVB alone. These synergistic effects of TWEAK+UVB on keratinocytes were likely due to upregulation of Fn14 expression by UVB. Our data strongly implicate TWEAK/Fn14 signaling in the pathogenesis of the cutaneous manifestations of lupus.
Reciprocal interaction between NK cells and DCs regulates the Th17 response by controlling the innate IFN-γ/IL-27 axis (BA13P.124)
Authors: Chong Wai Po , Chen Jun , Silver Phyllis , Caspi Rachel 
Published: January 2023

Abstract
IFN-γ is a pathogenic cytokine involved in inflammation. Paradoxically, its deficiency exacerbates experimental autoimmune encephalomyelitis, uveitis and arthritis. We previously showed that it is innate IFN-γ that is protective, whereas adaptive IFN-γ is pathogenic. Here we demonstrate by immunological and imaging methods, using IFN-γ-deficient mice repleted with IFN-γ-sufficient NK cells, that innate production of IFN-γ from NK cells is necessary and sufficient to limit autoimmunity and to reduce the number of IL-17A and GM-CSF producing effector T cells in the eye. Mechanistic studies demonstrated that, following immunization for uveitis, DC recruited IFN-γ-producing NK cells to the draining lymph node (DLN) and interacted with them in a CXCR3-dependent fashion. During the interaction, IFN-γ from NK cells induced DCs to produce IL-27, which in turn enhanced IFN-γ production by NK cells, forming a positive feedback loop. The NK-DC-dependent IL-27 inhibited development of the adaptive pathogenic IL-17 response and induced Tr1-like cells, which upon adoptive transfer ameliorated disease in an IL-10-dependent manner. Our data reveal that an early NK-DC interaction controls the adaptive Th17 response and limits tissue-specific autoimmunity through an innate IFN-γ/IL-27 axis.
Autoimmunity in interferon-gamma AU-rich element deleted-mice is associated with loss of marginal zone B cells and macrophages. (BA15P.217)
Authors: Hodge Deborah , Berthet Cyril , Coppola Vincenzo , Kastenmüller Wolfgang , Buschman Matthew , Shirota Hidekazu , Schaughency Paul , Anver Miriam , Reynolds Della , Sanford Michael , Kaldis Philipp , Young Howard 
Published: January 2023

Abstract
We generated a mouse model with a 162 nt AU-rich element (ARE) region deletion in the 3’ untranslated region (3’UTR) of the interferon (IFN)-gamma gene that results in chronic circulating serum IFN-gamma levels. Mice homozygous for the ARE deletion present both serologic and cellular abnormalities typical of patients with systemic lupus erythematosus (SLE). Marginal zone B (MZB) cells and marginal zone macrophages (MZMs) are absent in ARE-deleted (ARE-Del)-/- mice. ARE-Del+/- mice retain both MZB cells and MZMs and develop no or mild autoimmunity. However, low dose clodronate treatment in ARE-Del+/- mice specifically eliminates MZMs and promotes anti-DNA antibody development and glomerulonephritis. Our findings demonstrate the consequences of a chronic IFN-gamma milieu on MZB cell loss and MZM function in autoimmunity. Furthermore, similarities between disease states in ARE-Del-/- mice and SLE patients suggest that IFN-gamma may not only be a product of SLE but may be critical for disease onset and progression.
Sustained PI3K activation destabilized tTreg (BA15P.227)
Authors: Zhou Xuyu 
Published: January 2023

Abstract
Regulatory T cells (Treg) play a central role in immune homeostasis, whose dysfunction has been implicated in autoimmune disease pathogenesis. Two subsets Tregs have been defined in vivo: thymus derived tTregs and peripherally derived pTreg. The plasticity of Treg, especially the thymically derived tTreg remains considerable controversial. Since the conserved non-coding sequence 1(CNS1) of Foxp3 gene is only critical for the generation of induced pTreg cells, we generated a new BAC-transgenic mouse expressing a thy1.1 2A Cre fusion protein driven from the CNS1 deleted Foxp3 promoter (del CNS1 Foxp3-thy1.1-Cre). Characterization of these mice demonstrated the thy1.1 2A Cre fusion protein expressed exclusively in the Nrp1+Foxp3+ peripheral tTreg subset. Moreover, thy1.1 expression in the thymus was delayed compare with endogenous Foxp3. Stability of thy1.1+ Treg was tested by breeding the thy1.1 2A Cre Tg mouse line to a conditional YFP knock-in reporter mouse, termed ROSA26-stop-YFP. A subset range from 2% to 8% Cre mapped cells spontaneously lost Foxp3 expression in healthy animals, thus even the late developed tTreg cells have some extent of plasticity. Final, we found sustained PI3K activation by breeding with a conditional active PI3K Rosa26 knockin mouse, can significant destabilized late developed tTreg and generated pathogenic exFoxp3 cells in the homeostatic condition.
Development of alpha4beta1+MBP-speific T cells in the GALT (BA15P.225)
Authors: Ito Naoko , Yadav Sudhir , Mindur John , Ito Kouichi 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is a CNS autoimmune disease caused by activated myelin-specific T cells. To examine how Myelin Basic Protein (MBP)-specific T cells spontaneously differentiate into encephalitogenic T cells, we created transgenic mice that express MS-associated HLA-DR, DRB5*0101, and MBP-specific TCR isolated from an MS patient. We found that the MBP-specific T cells differentiated into GM-CSF+ Th1 and Th17 cells in the thymus spontaneously and migrate into the periphery including mesenteric lymph nodes and the gut-associated lymphoid tissue (GALT). Interestingly, we found that alpha4beta1 integrin was up-regulated in the GALT of spontaneous EAE mice although up-regulation of alpha4beta1 was not observed in mice that did not develop spontaneous EAE. Furthermore, reduction of gut commensal bacteria with antibiotics reduced the development of spontaneous EAE. This data suggest that commensal bacteria-mediated up-regulation of alpha4/beta1 integrin on the MBP-specific T cells in the GALT may be associated with the development of spontaneous EAE.
Exacerbated tissue homing of neutrophils during sepsis and TLR2-induced cytokine production are regulated by integrin α3β1. (CAM5P.230)
Authors: Lerman Yelena , Sarangi Pranita , Hyun Young-Min , Lim Kihong , Falkner Kathleen , Pietropaoli Anthony , Kim Minsoo 
Published: January 2023

Abstract
Neutrophil migration to sites of infection is vital for pathogen clearance, but massive neutrophil infiltration during systemic inflammation leads to tissue injury and host mortality. While β2 integrins are well known to mediate neutrophil firm adhesion and crawling, the role of β1 integrins in vascular extravasation has been less explored. In this study, we evaluated the surface expression kinetics of β1 and β3 integrin heterodimers on neutrophils during sepsis using mouse models (CLP and endotoxemia) and human samples. We show that only integrin α3β1 becomes significantly upregulated. Moreover, human subjects with sepsis, but not SIRS (systemic inflammation w/o infection) upregulated α3β1. The increased expression of α3β1 on neutrophils correlated with their activation state. Administration of synthetic peptide (LXY2), which binds at the ligand binding site on α3β1, and conditional gene deletion of the integrin in granulocytes significantly reduced the number of extravasated neutrophils and improved mouse survival. Host bacterial clearance remained unaffected. Interestingly, LXY2 binding to α3β1 on neutrophils was associated with a reduction in TLR2-, but not TLR4-induced IL-10 secretion in vitro. Previous studies suggest a role for IL-10 as a regulator of the transition from mild sepsis to irreversible septic shock. Thus, integrin α3β1 upregulation promotes enhanced neutrophil migration and, in turn, could regulate sepsis progression by modulating neutrophil IL-10 release.
CD13 is a novel regulator of TLR4 endocytosis in dendritic cells (CAM5P.241)
Authors: Ghosh Mallika , Subramani Jaganathan , Rahman Mamunur , Shapiro Linda 
Published: January 2023

Abstract
Toll like receptors (TLRs) recognize antigens produced by infection or injury to induce innate immunity via cell surface or endocytic pathways. TLR4 sequentially triggers both pathways, leading to TLR4 activation of the pro-inflammatory NF-kB pathway from the cell surface followed by dynamin-mediated TLR4 endocytosis, activation of TRIF and production of anti-inflammatory IRF (Interferon Regulatory Factor)3-regulated genes. We found that myeloid cell surface peptidase CD13 regulates TLR4 signal transduction by controlling dynamin-dependent endocytosis. In response to LPS, CD13KO DCs showed enhanced activation of IRF3, increase in its target gene IFN-β and inducible nitric oxide activity in a dynamin dependent manner. Endogenous TLR4 ligands produced in response to ischemic injury provoked hyperactivation of pIRF3 and its target cytokines in CD13KO animals. CD13KO DCs internalize higher levels of cell-associated TLR4 ligands released by dying cells or the TLR4 ligand LPS, indicating that CD13 regulates inflammatory TLR4 responses in vivo by modulating DC antigen uptake. Our results further suggest that CD13 and the LPS binding protein, CD14, may potentially act as opposing regulators of TLR4 endocytosis in inflammation. We propose that in response to injury or infection, CD13 serves to balance the immune response by modulating TLR4 induced pro- and anti-inflammatory cytokine profiles and maintain inflammatory equilibrium that is critical to proper wound healing.
Lymphocytes rapidly infiltrate into the brain following cardiac arrest and cardiopulmonary resuscitation (CA/CPR) (CAM5P.246)
Authors: Carter Jessica , Deng Guiying , Wagner David , Herson Paco 
Published: January 2023

Abstract
Isolated ischemic episodes, such as occurs with stroke, result in long-term neural damage. It is known that lymphocytes along with other immune cells infiltrate brain tissue promoting that process. During cardiac arrest, global ischemia then reperfusion occurs. Although inflammatory mechanisms have been linked to neuronal injury following global cerebral ischemia, the presence of infiltrating immune cells remains understudied. We examined the brains of mice at different time points after global cerebral ischemia induced by cardiac arrest and cardiopulmonary resuscitation (CA/CPR) and characterized the influx of lymphocytes into the injured brain. We observed that CA/CPR caused a rapid influx of lymphocytes within 3 hours of resuscitation that was maintained for the duration of our experiments. The large majority of infiltrating lymphocytes were CD4+ T cells that expressed the Th1 characteristic cytokines TNFα or INFγ. Of the CD4+ infiltrating cells, a large portion of them were also CD40+ (Th40). Brain injury models attribute much of the neural damage to microglia and subsequent oxidative stress. We found that the CA/CPR injury model was largely dependent upon T cells, and the lack of functional T cells in TCRα knockout mice significantly reduced neuronal injury. This data indicate that studies investigating the neuro-immune response after global cerebral ischemia should consider the role of infiltrating T cells in orchestrating the acute and sustained immune response.
Changes in plasma angiopoietin-like protein 2 levels and macrophage migration associated with hibernation in thirteen-lined ground squirrels (CAM5P.232)
Authors: Taylor Bernadette , Hanson Erika , Shaheen Bayan , Cooper Scott 
Published: January 2023

Abstract
Thirteen-lined ground squirrels in the northern United States hibernate from around late September until March. In late summer they gain a substantial amount of fat that is gradually expended as fuel over the course of hibernation. Immune system changes during hibernation include a 90% drop in circulating leukocytes and a reduction in immune responsiveness. Adipose tissue in obese humans is associated with changes in leukocyte migration including macrophage infiltration. This study explored an adipose-associated inflammatory protein, angiopoeitin-like protein 2 (ANGPTL2) as a potential mediator of leukocyte migration in hibernating ground squirrels. Plasma ANGPTL2 in eight ground squirrels was measured by quantitative Western blot at non-hibernating, entering hibernation, hibernating, interbout arousal and post arousal time points. ANGPTL2 levels were higher in hibernation than all other time points and were significantly higher in hibernation compared to interbout arousal or post arousal time points. Macrophages were detected in adipose tissue of hibernating ground squirrels using a ground squirrel macrophage specific monoclonal antibody in immunoperoxidase stained frozen sections. Macrophage-staining in the spleen appeared to be unusually dense in venous sinuses of hibernating compared to non-hibernating squirrels.
Super-hydrophilic nanostructured titanium surface improves implant osseointegration by mediating macrophage polarization (CAM5P.233)
Authors: Chen Lihua , Ma Qianli , Fang Liang , Wang Ying , Liu Rongrong , Zhang Yusi , Jin Boquan , Zhang Yumei 
Published: January 2023

Abstract
Aims: The purpose of this study was to fabricate a nanostructured surface on a titanium (Ti) implant that can regulate the macrophage inflammatory response and thereby improve the performance of the implant. Materials and Methods: Pure Ti samples were anodized and then subjected to UV irradiation to generate super-hydrophilic, nanostructured TiO2 surfaces. Their surface characteristics were analyzed, and osteointegration as well as the inflammatory response of modified implants were evaluated in vivo and in vitro. Results: Both nanostructured TiO2 surfaces showed super-hydrophilic and multi-tubular surface topography with diameters of 30 nm (NT5) and 80 nm (NT20). Although the in vitro osteogenic activity of bMSCs was similar on the two surfaces, the NT5 surface showed more bone formation, less inflammation, and a reduced CD68+ macrophage distribution in vivo compared to the NT20 and polished Ti surfaces. Further experiments revealed that the NT5 surfaces suppressed macrophage secretion of inflammatory cytokines and induced M2 polarization, which is associated with improved healing. By contrast, promoted M1 polarization could further impair bone regeneration. Conclusion: The results demonstrate the dominant role of macrophage-related inflammation in bone healing around implants and that nano-scale surface topography may have an immune-regulating effect that can support the success of implants.
IL-13 heteroreceptor and Th1 cells: the race for control of neonatal immunity (CCR5P.256)
Authors: Miller Mindy , Dhakal Mermagya , Chen Weirong , Ellis Jason , Khairallah Marie-Therese , Ukah Tobechukwu , Zaghouani Habib 
Published: January 2023

Abstract
Neonates are susceptible to microbial infections because they lack Th1 cells, the major effectors against microbes. Also, newborns are prone to allergic reactions perhaps due to their tendency to develop Th2 biased immunity. Fine analysis of primary neonatal immunity indicated a balance among Th1 and Th2 cells but the former display an unusual expression of IL-13 receptor alpha 1 (IL-13Rα1). Furthermore, this chain heterodimerizes with IL-4Rα and the resulting IL-4α/IL-13Rα1 provides a death receptor through which IL-4 of Th2 cells drives apoptosis of Th1 counterparts during secondary immunity. Herein, we observed that IL-4 and IL-13 signaling through the heteroreceptor shifts antigen-induced activation towards the mitochondrial apoptotic pathway by recruiting interferon-regulatory factor 1 (IRF-1). Further investigation of heteroreceptor signaling of neonatal Th1 cells should yield information useful for the development of pediatric vaccines and drugs against allergies.
IL-23 Maintains Tc17 CD8 T Cell Phenotype but not Commitment to IL-17 production (CCR6P.282)
Authors: Yen Hung-Rong , Lin Bo-Shiou , Huang Wan-Chun , Liao Cha-Hui , Yeh Yueh-Chiao , Liang Kai-Li 
Published: January 2023

Abstract
CD8 T cells that produce IL-17 is an unknown field with lots to explore. In our previous studies, we found a novel subset of IL-17 producing CD8 T cells persisted longer in a self-antigen autoimmune model versus conventional IFN-g producing Tc1 cells; furthermore, Tc17 can convert to an IFN-g producing phenotype in vivo. In the present study, IL-17 producing Tc17 cells were purified by the cytokine-secretion assay and re-stimulated with anti-CD3/CD28 monoclonal antibodies with exogenous IL-12 or IL-23 cytokine. We found that IL-23 maintains IL-17-producing Tc17 CD8 T cell phenotype. However, administration of IL-23 is not enough to maintain the Tc17 phenotype. Addition of other cytokines such as IL-12 repressed IL-17 production and gene expression. In summary, IL-23 can maintain the Tc17 phenotype, but cannot promote the commitment of Tc17 CD8 T cells to an IL-17-secreting lineage.