bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mincle signaling contributes to the inflammatory response after traumatic brain injury (P1297)
Authors: de Rivero Vaccari Juan Pablo , Brand III Frank , Berti Aldo , de Rivero Vaccari Juan Carlos 
Published: January 2023

Abstract
The inflammatory response after traumatic brain injury (TBI) contributes to neuronal death and progressive axonal loss to areas undamaged by the initial trauma. Moreover, anti-inflammatory therapies offer great hope to improve outcomes after TBI. Thus, it is important to find therapeutic targets to better control the inflammatory response after injury. In this study we used cells in culture, an animal model of TBI as well as brain and CSF samples obtained from patients with TBI to investigate, after injury, the role of the CLR family member, macrophage-inducible C-type lectin (mincle) in the production of the inflammatory cytokine TNF. Here we report that mincle is activated after TBI in both, humans and rodents. Our data show that moderate fluid percussion brain injury (using male Sprague-Dawley rats (250-350 g) subjected to moderate fluid-percussion brain injury (1.8-2.2 atm)) is associated with increase expression of SAP130, which activates mincle signaling resulting in phosphorylation of the enzyme syk. Moreover, we identified increased expression of SAP130 and mincle in cortical neurons after TBI. In addition, we show that SAP130 stimulates mincle signaling in primary neuronal cultures, resulting in increase expression of the inflammatory cytokine TNF. Acute TNF expression after TBI is associated with deleterious outcomes. In conclusion, these findings suggest mincle as a novel therapeutic target that can be used to lower the levels of TNF after CNS injury.
MicroRNA-let-7a promotes E2F-dependent proliferation in mouse macrophages and mesangial cells (P1301)
Authors: Chafin Cristen , Regna Nicole , Caudell David , Reilly Christopher 
Published: January 2023

Abstract
Purpose: Cell proliferation is a common histological feature of lupus nephritis (LN) in the autoimmune disease systemic lupus erythematosus (SLE). MicroRNAs (miRNAs) are small, non-coding RNAs that post-transcriptionally regulate gene expression. In this study, we used in vitro approaches to investigate cell proliferation through E2F-mediated activation of miRNA-let-7a. Methods: A mouse macrophage and a mouse mesangial cell line were transfected with let-7a mimics or a non-targeting control and treated with control or stimulating media. Cell cycle distribution was measured by flow cytometry. Proliferation was measured by MTT assay. Cells were transfected with E2F2 siRNA or a non-targeting control. Let-7a and E2F2 expression were measured by real-time RT-PCR. NFκB activation and IL-6 protein levels were measured by ELISA. Results: Let-7a decreased the number of stimulated cells in G0-G1 phase, resulting in an increase in S and G2-M phase. Cell proliferation was increased in stimulated cells transfected with let-7a. Let-7a significantly increased NFκB activation and E2F2 mRNA expression in stimulated cells. Silencing E2F2 significantly decreased let-7a expression and IL-6 production in stimulated cells. Conclusion: Let-7a induces macrophage and mesangial cell growth and proliferation, in part by activating the transcription factors NFκB and E2F2. This positive feedback loop may explain one mechanism of hyperplasia in lupus.
Cooperation of CD86 signaling with TLR-2 in the activation of resting B cells: a novel BCR independent pathway of stimulating B cells (P1126)
Authors: Chodisetti Sathi Babu , Jain Shweta , Agrewala Javed 
Published: January 2023

Abstract
The molecules of adaptive and innate immunity cooperate with each other in inducing effective immune response. It is well established fact that B cells can be activated through BCR mediated signaling. Here, we report an alternative strategy of activating resting B cells (RB cells) through concurrent engagement of CD86 and Toll like receptor-2. Concomitant signaling through CD86 and TLR-2 significantly augmented the longevity and activation profile of RB cells. Further, these cells showed enhanced pinocytosis and receptor mediated endocytosis. Furthermore, these cells secreted high levels of IgM and IgG1 and showed better ratio of surface IgG/IgM. Moreover, this process of activation amplifies the population of marginal zone precursors, which connect innate and adaptive arms of immune response. There was also upregulation of set of genes involved in the B cell activation, proliferation, maturation and endocytosis. The data are well in agreement with the gene expression profiles and insinuates to further decipher the molecular mechanism behind this concurrence. An insight into the detailed mechanism may pave way to design vaccines based on this signaling strategy.
A unique DNA methylation signature defines human CD4 memory T cells (P1148)
Authors: Komori Heather , Hart Glenn , LaMere Sarah , Mark Adam , Salomon Daniel 
Published: January 2023

Abstract
Memory CD4 T cells are “primed” for rapid responses to antigen; however, the molecular basis of this “priming” is incompletely understood. Methylation of CpG dinucleotides in gene promoters is an epigenetic modification known to regulate gene transcription. Using a new protocol we developed for targeted bisulfite sequencing (Genome Res, 2011), we assessed promoter methylation in human naïve and memory CD4 T cells of 2100 genes (56,000 CpG) linked to activation by deep RNAseq. 466 individual CpGs representing 132 genes are differentially methylated between resting naïve and memory cells. 21 genes exhibited both differential methylation and gene expression before activation, linking promoter DNA methylation states to gene regulation; 6 genes encode proteins closely studied in T cells while 15 genes represent novel targets for further study. 39 genes exhibited reduced methylation in resting memory cells coupled with increased gene expression with activation compared to naïve cells including 8 chemokines and cytokines. These data reveal specific genes more rapidly expressed in memory compared to naïve cells and regulated by DNA methylation. Our findings define a DNA methylation signature unique to memory CD4 T cells and correlated with activation-induced gene expression. We are just completing a new analysis of naïve vs. memory at 3 time points (day 0, 5, 14) using a new protocol for genome-wide CpG capture and deep bisulfite DNA sequencing to validate and extend this work.
Control of the molecular behavior of SHIP by FcγRIIB and Nck (P1122)
Authors: Pauls Samantha , Ray Arnab , Marshall Aaron 
Published: January 2023

Abstract
Leukocyte activity is controlled by balancing activating and inhibitory signals in order to generate effective immune responses while avoiding tissue damage and autoimmunity. This project examines an important regulatory circuit in B cells that involves the lipid phosphatase SHIP. Localization, phosphorylation, and binding to interaction partners are important factors in the regulation of this protein, however knowledge of their inter-relationships is incomplete. Here, we examine the influence of two interaction partners on the molecular behaviour and function of SHIP. First, we examine in detail how the inhibitory receptor FcγRIIB controls SHIP localization dynamics. We have observed that co-engagement of FcγRIIB along with the B cell receptor in A20 cells does not influence the magnitude or kinetics of SHIP-EGFP recruitment to the membrane as assessed by confocal microscopy, however it does alter mobility at the cell periphery as measured by fluorescence recovery after photobleaching. Next, we probe the influence of a novel binding partner, the adaptor protein Nck. We have demonstrated an interaction between SHIP and Nck by both Biacore affinity analysis and pull-down assays. Functional relevance will be addressed through mutagenesis experiments. With these aims we hope to enhance our understanding of an interaction that is already recognized as relevant and investigate the function of a novel interaction that occurs at a previously uncharacterized regulatory site.
Tregs suppress CD8 T cell effector function in inflamed tissue in a mouse model of autoimmune diabetes (P1018)
Authors: Mahne Ashley , Chou Annie , Tang Qizhi 
Published: January 2023

Abstract
The aim of this study was to determine the means by which Tregs control disease in the NOD mouse model of autoimmune diabetes. Treatment of pre-diabetic mice with ongoing islet destruction with islet antigen-specific Tregs expanded from BDC2.5 TCR transgenic mice led to a rapid restoration of insulin production and a concomitant selective downregulation of a cytotoxic T lymphocyte signature, which included CD8α, IFNγ, granzymes A and B, and Cxcl9. This downregulation could be partially attributed to a decrease in intra-islet CD8 T cell numbers due to impaired recruitment of CD8 T cells to the islets. In addition, Treg treatment also reduced effector function of persisting CD8 T cells. Two-photon imaging showed that Tregs did not significantly disrupt interactions between islet CD8 T cells and dendritic cells, but abrogated IFNγ production by CD8 T cells. This suppression appeared to take place at multiple stages of CD8 T cell differentiation. Tregs inhibited differentiation of newly recruited CD8 T cells to IFNγ producers completely, but prevented IFNγ protein expression without affecting IFNγ message in pre-activated CD8 T cells. Altogether, these results indicate that CD8 T cells are the primary early targets of Treg control of tissue inflammation and rapid disease resolution by Tregs can be attributed to efficient control of CD8 T cell infiltration, reactivation in the tissue, and their expression of effector functions.
The innate-like T cells are required to modulate acute inflammatory response (P1050)
Authors: Tang Hong , Feng JIn , Zhang Chao 
Published: January 2023

Abstract
Macrophages infiltration and activation in myocardium is a pivotal immunopathological lead to hypertensive cardiac micro-injury, but underlying mechanisms remain elusive. We have found that CD8 KO or CD8+ T cells depletion by antibody significantly reduce cardiac pro-fibrotic inflammatory responses induced by angiotensin II (Ang II) infusion, whereas CD8 KO mice reconstituted with CD8+ T cells became sensitive. More importantly, CD8+ T cells are required for macrophage infiltration in myocardium and subsequent activation to express pro-inflammatory cytokines and chemokines, including MCP-1. Furthermore, transwell experiments showed that macrophage activation requires direct contact with activated CD8+ T cells, but with TCR dispensable. TCR-independent activation of macrophage is further confirmed in vivo, where OT-I transgenic mice show a similar cardiac pro-inflammatory response to Ang II as wt mice. Finally, IFNγ seems required for influx and activation of CD8+ T cells in myocardium in response to Ang II, that subsequently activate macrophages in the onset of cardiac inflammation. Thus, TCR-independent innate nature of CD8+ T cells is both necessary and sufficient for macrophage-induced hypertensive cardiac fibrosis. In conclusion, TCR-independent activation of macrophages by CD8+ T cells casts yet a novel innate function of T cells that is required to activate inflammatory response of macrophages to danger signals.
Choline acetyltransferase+ T cells represent a discrete lymphocyte subset that regulates innate immunity. (P1008)
Authors: Olofsson Peder , Oswald Michaela , Hanes William , Lu Ben , Chavan Sangeeta , Gregersen Peter , Tracey Kevin 
Published: January 2023

Abstract
Neural circuits regulate cytokine production and inflammation. A vagus nerve circuit inhibits TNF production by acetylcholine signaling through the α7 nicotinic acetylcholine receptor on macrophages in spleen, but spleen nerves lack the machinery for acetylcholine biosynthesis. Instead, the nerve signal is relayed by CD4+CD44hiCD62Llow choline acetyltransferase+ (ChAT+) T cells, which produce the required acetylcholine. Understanding of the ChAT+ T cell phenotype is incomplete. mRNA levels in ChAT+ and ChAT- splenic T cells were measured by gene array. Hierarchical clustering of significantly different transcripts rendered four clusters. Expression data from the ChAT+/ChAT- dataset was also analyzed together with publicly available data on immune cell mRNA expression from the ImmGen consortium. In principal component analysis of this dataset, NK-cells, dendtritic cells/macropages, B cells and T cells formed separate clusters. ChAT+ T cells clustered among T cells, but separately from other T cell subpopulations, and were distinguished by the first principal component. Gene ontology analysis of this component showed enrichment of the G-protein coupled receptor signaling pathway, proliferation, and metabolism terms. In vitro, presence of ChAT+ T cells inhibited macrophage TNF production. Taken together, these data indicate that ChAT+ T cells are phenotypically distinct from other cell types in the ImmGen dataset and suggests that ChAT+ T cells are a novel T cell subset.
Host resistance to a gastrointestinal parasite regulated by Mina, an immunoregulatory member of the JmjC protein family (P1014)
Authors: R.Pillai Meenu , Van Wyhe Renae , Vogel Peter , Glaysher Bridget , Coleman Mathew , Ratcliffe Peter , Coles Mark , Bix Mark 
Published: January 2023

Abstract
T helper (Th) 1 and Th2 responses while critical for clearance of pathogens, when dysregulated results in autoimmune disease or allergy and asthma respectively. The genetic predisposition of certain inbred mouse strains towards developing Th2 as opposed to Th1 response (Th2-bias) has long been associated with differential susceptibility across a wide range of immunological diseases. However, its molecular genetic basis and thus the causal relationship to disease states are poorly understood. Prior investigations into the nature of this relationship resulted in the physical isolation of Dice1.2 on chromosome 16 and identification of Mina as a Dice1.2 resident gene exhibiting Il4 repressor activity. Thus, we hypothesized that Mina’s physiological role is to negatively regulate Th2 responses. To test this, we generated Mina KO mice and studied their response to the intestinal nematode T. muris. We found that Mina KO mice are hyper-resistant to T. muris infection. Surprisingly, the elevated T. muris resistance of Mina KO mice was not associated with an increased Th2 response but a defect in Th1 response. This correlated well with reduced inflammation in the cecum and reduced expression of antigen specific serum IgG2a. Further analysis revealed diminished expression of epithelial cell-specific Relmβ in Mina KO mice. These results highlight the importance of Mina in immune regulation and provide insight into the complex immune network perturbed by its expression and function.
Enhancement of vaccine-induced respiratory syncytial virus-specific memory CD8 T cell responses by targeting 41BB (P6158)
Authors: Lee Sujin , Moore Martin 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) causes significant morbidity and mortality in children and the elderly. There is no RSV vaccine in use. One of the major obstacles to RSV vaccine development is that RSV re-infects humans throughout life due to lack of durable protective immunity. We hypothesized that maintenance of robust and long-lasting memory CD8 T cells would be the key factor to prevent RSV infection. Recently, we generated a TriVax vaccination approach (peptide, polyI:C, and agonistic α-CD40 mAb). TriVax vaccination in mice completely protected against RSV infection at the effector phase, but protection against RSV was partial during the memory phase. 41BB (CD137) plays a key role in the survival of memory CD8 T cells. To determine whether co-administration of α-41BB in conjunction with TriVax could improve the maintenance of RSV-specific CD8 T cells at memory phase, we administered α-41BB (clone 3H3) or control IgG on the day of TriVax vaccination, and again 2 days later. Data indicated that co-administration of α-41BB and TriVax maintained 30% RSV-specific CD8 T cells until day 35 after the boost. Administration of TriVax+α-41BB followed by challenge using RSV rA2-line19F strain resulted in increased CD8 T cell cytokine expression and protection against RSV infection at memory phase as compared to TriVax+isotype control. Collectively, these data suggest that 41BB may be a promising candidate for the induction of long-lasting RSV-specific memory CD8 T cell immunity.
Dissecting the potential of resident CD11c-positive and CD11c-negative cells to support vitamin A-driven virus-specific immune responses in the upper respiratory tract (P6139)
Authors: Rudraraju Rajeev , Jones Bart , Surman Sherri , Sealy Robert , Woodland David , Hurwitz Julia 
Published: January 2023

Abstract
Morbidity and mortality due to viral infections are major health concerns, particularly when individuals are vitamin A (VitA) deficient (VAD). Previous literature has suggested that CD11c+ dendritic cells in the gastrointestinal tract are largely responsible for VitA metabolism and the associated homing and function of gut lymphocytes. To determine if/how VitA impacts virus-specific immunity in the upper respiratory tract (RT), we administered virus intranasally (IN) to VAD mice. We found that virus-specific B- and T-cell responses were altered, with a significant down-regulation of IgA in the nasal wash of VAD mice compared to controls. We next asked if RT cells could autonomously support VitA-driven IgA production. We harvested CD11c+ and Cd11c- cells from diffuse nasal associated lymphoid tissues and found that both expressed RALDH mRNA, as required for VitA metabolism. Furthermore, preliminary data showed that both populations enhanced IgA in activated splenocyte cultures. To investigate which CD11c- cells were pertinent to function, we tested a RT epithelial cell line. These cells: (i) expressed RALDH mRNA and (ii) enhanced IgA in the presence of VitA A derivatives. In total, data highlighted the autonomous potential of RT cells to support a VitA-driven immune response. Results may ultimately instruct a clinical healthcare initiative to supplement VAD individuals with VitA by the oral or IN route, to enhance RT immunity toward viral infections and viral vaccines.
The DNA sensing inflammasome, AIM2, is critical for protection from influenza infection (P6140)
Authors: Schattgen Stefan , Kurt-Jones Evelyn , Fitzgerald Katherine 
Published: January 2023

Abstract
Sensing of viral-derived nucleic acids following entry into cytosol of the host cell triggers inflammasome activation leading to maturation of interleukin 1β (IL-1β) and IL-18. In addition to activating the inflammasome directly within infected cells, viral infection can cause extensive tissue damage resulting in the accumulation of endogenous danger-associated molecular patterns (DAMPs), including self-DNA, which may provide signals in triggering inflammasome activation in neighboring uninfected cells. Though several studies have shown inflammasome activation within infected cells, it is unknown whether the sensing of DAMPs by uninfected bystander cells and the subsequent inflammasome activation provides an important proinflammatory signal in establishing an effective antiviral response. Using mice deficient in the DNA-sensing inflammasome component AIM2, we demonstrate AIM2 is required for protective immunity to influenza A virus (IAV), an orthomyxovirus with an ssRNA genome and a strictly RNA replication cycle. In addition to their increase in mortality and morbidity, Aim2-deficient mice show a defect in levels caspase-1-dependent cytokines IL-1β and IL-18, both important in the activation of cellular antiviral responses, following IAV challenge. Collectively, these data findings suggest that the sensing of self-DNA by AIM2 plays a critical role in triggering innate immune responses following infection with an RNA virus.
Stay off my lawn: T cell territorialism in virus-infected tissues of the respiratory tract (P6150)
Authors: Hurwitz Julia , Sealy Robert , Surman Sherri , Thomas Paul , Dayton Barry , Rudraraju Rajeev 
Published: January 2023

Abstract
Little is known of the population dynamics that influence lymphocyte positioning in virus- infected tissues. To address this issue, we primed animals with Sendai virus (SeV) and harvested cervical lymph nodes (CLN) or spleens 10 days later. Cells were unenriched or enriched for CD3, CD4 or CD8 phenotypes and transferred intraorbitally to GFP-SeV infected mice. Cell positions were then examined by confocal microscopy. Results showed that donor cells were configured in infected tissues with limited overlap. In most infected fields, the mean distance between a donor cell and its nearest neighbor was greater than would be predicted by random uniform distribution. To explain these findings, we hypothesized that a newly resident lymphocyte may instruct expression of unique chemokines/cytokines (c/c) in its immediate vicinity to mark its territory and inhibit co-localization by other cells. In vitro tests supported this hypothesis in that when virus-primed CLN were co-cultured with stroma and SeV, there was a synergistic increase in c/c production. Transwell experiments then showed that cell migration was intricately affected by the c/c milieu at local and distant sites. We therefore propose that a newly-resident lymphocyte in an infected tissue creates a c/c halo to identify and claim its territory. Lymphocyte cross-talk may ensure a spatial distribution of effectors to avoid redundancies and enhance the efficiency with which effector cells clear their virus infected targets.
Exhaustion of influenza-specific CD8+ T cells during primary infection leads to severe disease (P6152)
Authors: Rutigliano John , Morris Melissa , Oguin Thomas , McClaren Jennifer , Doherty Peter , Thomas Paul 
Published: January 2023

Abstract
It remains unclear why some strains of influenza induce severe disease while others cause more mild illness. We approached this question by investigating the CD8+ T cell response in a mouse model system comparing high and low pathological influenza infections. We discovered that early viral titer was not the determining factor in the outcome of disease. Instead, increased numbers of antigen-specific CD8+ T cells and elevated effector function on a per cell basis were found in low pathological infection and correlated with reduced illness and later time point viral titer. High pathological infection was associated with increased PD-1 expression on influenza-specific CD8+ T cells, but blockade of PD1-L in vivo did not rescue severe disease, and CD8+ T cell effector function was not restored. However, after anti-PD-1L treatment CD8+ T cell numbers were dramatically increased in high pathological infection, but not in low pathological infection. Additional studies in IL-6-deficient mice and WT mice treated with anti-G-CSF antibody showed a partial reversal of the severe disease phenotype, but these treatments failed to fully rescue severe disease. These data suggest that high pathological acute influenza infection is associated with CD8+ T cell exhaustion and an attendant increase in inflammation that was observed in the airway microenvironment during the early stages of infection. As a result, therapeutic approaches specifically aimed at modulating airway inflammation may promote efficient CD8+ T cell activity, with improved morbidity and mortality of severe influenza infections.