bluuebunny/crossref_metadata_2025_split huggingface dataset:
Elucidating a role for a hematopoietic-tropic microRNA in T cell (IRM12P.650)
Authors: Menoret Antoine , Kumar Sanjeev , Palakodeti Dasaradhi , Graveley Brenton , Vella Anthony 
Published: January 2023

Abstract
To gain new insight into how miRNA impact T cell responses, mice were immunized with costimulation and adjuvant to generate potent T effector responses. We generated a cDNA library from lymphoid tissue. Using sequencing of size-fractionated cDNA we cloned 6 potential new miRNAs with characteristic stem loop folding. One of these candidates, termed miR-R89 but not yet registered in miRBase, was unique since its expression was hematopoietic specific and regulated during immune activation. The targets and function of miR-R89 are currently unknown, but our preliminary data show it is expressed in a T cell clone known to cause experimental autoimmune encephalitis. Transfection with a locked nucleic acid (LNA) mimic of miR-R89 in EL4 thymoma cells increased IL-2 secretion after PMA-ionomycin (PI) stimulation compared to transfection with control LNA. RNAseq analysis of the transfected cells revealed that mRNA expression of two transcription factors involved in metabolism, Tfe3 and Tef, was reduced by miR-R89 mimic. Interestingly, 3’UTR mRNA of Tfe3 and Tef have several theoretical binding sites for miR-R89. Treatment of PI-stimulated EL4 cells with UK5099, an inhibitor of mitochondrial pyruvate transport and metabolism, mediated a similar increase of IL-2 secretion than observed after transfection with miR-R89. These data suggest the putative miRNA miR-R89 could target Tfe3 and Tef mRNA and function as a metabolic regulator of T cell activation.
The level of antigen expression in the liver predicts the fate and functional outcome of antigen-specific CD8 T cells (LYM5P.706)
Authors: Bertolino Patrick , Wong Yik Chun , Tay Szun Szun , McDonald David , Sierro Frederic , Roediger Ben , Wood Nicole , McGuffog Claire , Bremner Kate , Alexander Ian , Weninger Wolfgang , McCaughan Geoffrey , Bowen David 
Published: January 2023

Abstract
A wide spectrum of CD8 T cell responses can be elicited against antigens (Ag) expressed in the liver, ranging from tolerance to full effector function. Identifying the factors involved would allow us to predict the outcome of liver transplantation and viral hepatitis. To investigate how CD8 T cells respond to liver Ags, we have developed recombinant adeno-associated viral vectors that, when injected into mice, allow de novo Ag expression in a variable number of hepatocytes. Within the first day of Ag expression by at least 70% of hepatocytes, most naïve Ag-specific CD8 T cells were activated in the liver via direct presentation by hepatocytes. CD8 T cell activation in lymph nodes and spleen via cross presentation was detected a day later, suggesting that primary activation of CD8 T cells against hepatocyte-expressed Ag is compartmentalised. These T cells proliferated, developed into CTLs in the first week but were unable to kill all Ag-expressing hepatocytes. Most effector T cells were rapidly deleted while those surviving became exhausted and unresponsive over time. In contrast, when less than 10% of hepatocytes expressed the targeted Ag, CD8 T cell were gradually activated in liver and lymphoid tissues but were not deleted. Instead, they eliminated all Ag-expressing hepatocytes and maintained their effector function overtime. In summary, this study shows that intrahepatic Ag load dictates both the fate and function of CD8 T cells recognising hepatocyte-expressed Ags.
Resveratrol regulates the miR-124/sphingosine kinase 1 axis in encephalitogenic T cells and protects from experimental multiple sclerosis (LYM5P.709)
Authors: Gandy K. , Nagarkatti Mitzi , Nagarkatti Prakash 
Published: January 2023

Abstract
Resveratrol (RES) is a naturally-occurring phytoestrogen with anti-inflammatory properties. In this study, we used a mouse model of multiple sclerosis, experimental autoimmune encephalitis (EAE), to evaluate the effect of RES on T helper cell function. We hypothesized that RES protects against EAE via regulation of encephalitogenic T cell miRNA expression. We found that RES treatment diminished clinical parameters of EAE and altered immune cell composition and activation in the brain. Analysis of brain-derived CD4+ T cells identified significant up-regulation of several miRNAs with RES treatment. In silico analysis of target genes revealed overlap with cell growth and proliferative functions. We identified brain-specific alteration of the miR-124/sphingosine kinsase 1 (SK1) axis in RES-treated mice, with no significant changes in peripheral CD4+ cells. In vitro RES treatment of peripherally-derived mononuclear cells had no effect on miR-124 expression, supporting the brain-specific regulation of CD4+ T cell miRNA expression by RES. Lastly, we evaluated the consequence(s) of RES-mediated alteration of the miR-124/SK1 axis by assessing encephalitogenic CD4+ T cell function. The current study demonstrates a role for the miR-124/SK1 axis in RES-mediated protection from EAE, suggesting that RES may be an ideal candidate for the treatment of neuroinflammatory diseases.
Calmodulin and PI(3,4,5)P3 cooperatively bind to the Itk Pleckstrin homology domain to promote efficient calcium signaling and IL-17A production in T cells (LYM5P.710)
Authors: Huang Yina , Wang Xinxin , Boyken Scott , Shea Madeline , Andreotti Amy 
Published: January 2023

Abstract
Precise regulation of the kinetics and magnitude of Ca2+ signaling enables this signal to mediate diverse responses, such as cell migration, differentiation, vesicular trafficking, and cell death. Here, we showed that the Ca2+-binding protein calmodulin (CaM) acted in a positive feedback loop to potentiate Ca2+ signaling downstream of the Tec kinase family member Itk. Using NMR (nuclear magnetic resonance), we mapped CaM binding to two loops adjacent to the lipid-binding pocket within the Itk pleckstrin homology (PH) domain. The Itk PH domain bound synergistically to Ca2+/CaM and the lipid phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P3], such that binding to Ca2+/CaM enhanced the binding to PI(3,4,5)P3 and vice versa. Disruption of CaM binding attenuated Itk recruitment to the membrane and diminished release of Ca2+ from the endoplasmic reticulum. Moreover, disruption of this feedback loop abrogated Itk-dependent production of the proinflammatory cytokine IL-17A (interleukin-17A) by CD4+ T cells. Additionally, we found that CaM associated with PH domains from other proteins, indicating that CaM may regulate other PH domain-containing proteins.
Determining the contribution of antigen-specific, CD4+ recent thymic emigrants to Salmonella persistence in vivo (LYM5P.661)
Authors: Goggins J. , Kurtz Jonathan , Israel Kate , McLachlan James 
Published: January 2023

Abstract
Typhoid fever, caused by infection with Salmonella bacteria affects approximately 21 million people annually, killing nearly 200,000. Recent thymic emigrants (RTEs) are the youngest subset of peripheral T cells, which are known to differ in how they functionally contribute to the naïve T cell pool. This distinctive cell population is known to contribute to the maintenance of T cell receptor diversity in the peripheral T cell pool, but their involvement in combating chronic bacterial infections, such as typhoid fever, has not been explored. In the present study, we hypothesized that RTEs are essential contributors to the CD4 T cell response during persistent Salmonella Typhimurium (ST) infection, which predominantly relies on helper T cell immunity to control bacteria. Here, we show that thymectomy of adult mice prior to oral infection with ST led to significantly increased bacterial burdens in the spleen, mesenteric lymph nodes and liver 30 days post-infection compared to sham surgical controls, suggesting Salmonella-specific RTEs may contribute to bacterial clearance. Surprisingly, existing peripheral Salmonella-specific T cells also increased significantly in number, implying that these cells are somehow incapable of controlling infection in the absence of RTEs. The results of this study provide insight into how new pathogen-specific T cells leaving the thymus participate in combatting a persistent bacterial infection.
Effects of natural supplementation on human T cell longevity pathways (LYM6P.723)
Authors: Valenzuela Hector , Casarez Nicholas , Hom Lisa , Daoudi Sarah 
Published: January 2023

Abstract
Nutraceuticals are natural compounds that contain both nutritional as well as medicinal benefits. Various nutraceuticals (curcumin and resveratrol) obtained from plant extracts have been shown to improve fitness and extend lifespan by various mechanisms. In addition other compounds have also been shown to extend lifespan (rapamycin, spermidine and nicotinamide mononucleotide). Although the end result for these compounds maybe extension of lifespan, comparison of their signaling pathways remains to be determined. The objective of our research was to compare the signaling pathway of these compounds on both normal human T cells and Jurkat cell lines. We used RT-PCR to measure gene expression of NF-Κβ, AMPK, sirtuin 1-7 and autophagy specific genes (ATG5 and MAPILC3B/LC3) for all of these compounds. In addition, we show a direct correlation between concentrations of treatment and the upregulation of gene expression to the viability of the cells.
STAT3 regulates iNKT cell development and function but is not required for NKT17 cell development (LYM6P.715)
Authors: Mi Qing-Sheng , Zhang Xilin , Li Guihua , Koralov Sergei , Zhou Li 
Published: January 2023

Abstract
Signal transducer and activator of transcription 3 (STAT3) controls the expansion of T help 17 cell (Th17) lineage through induction of transcription factor orphan nuclear receptor (RORγt). CD1d-restricted invariant natural killer T (iNKT) cells are potent regulators of diverse immune responses and have three sublineages, NKT1, NKT2, and NKT17 cells. However, it still remains unclear how to regulate NKT17 cell development. Using lymphocyte-specific STAT3 hyperactivation mice, we found that overactivation of STAT3 universally suppressed iNKT cell cytokine secretion, including IL-17, while it enhanced conventional Th17 cell development. Consistently, STAT3 hyperactivation reduces the RORγt expression of iNKT cells. Surprisingly, bone marrow chimera experiments revealed that STAT3 overactivation disturbed iNKT cell maturation and increased IFNγ- and IL-4-secreting iNKT cells while it did not affect IL-17 secreting iNKT cells and did not alter RORγt expression of iNKT cells. Our results demonstrate that STAT3 regulates the development and function of iNKT cells, but unlike conventional Th17 cells, STAT3 is not required for IL-17 production and RORγt expression in iNKT cells.
A role for dectin-1 in the CNS during chronic Toxoplasma gondii infection (MPF5P.739)
Authors: Worth Danielle , Nance John Philip , Jang Jessica , David Clement , Nair Meera , Wilson Emma 
Published: January 2023

Abstract
The latent stage of T.gondii infection is characterized by the formation of tissue cysts within the CNS. These tissue cysts are formed by manipulation of the parasitophorous vacuole, and chitin is a structural component of the cyst wall. Our lab has shown that Toxoplasma cysts induce an M2 phenotype in macrophages in a contact-dependent manner. Upon contact, these macrophages actively secrete the mammalian chitinase, AMCase, to break down the cyst wall. We have previously demonstrated that AMCase production by macrophages is essential for control of cyst burden in the brain during T. gondii infection. The molecular interactions involved in macrophage-cyst recognition, and the signaling pathway for AMCase production are as yet unknown. Although chitin recognition has been poorly defined, studies suggest dectin-1 may be a receptor. Here, we present data on a role for dectin-1 during T. gondii infection. Dectin-1 is upregulated in the brain in response to infection and is expressed on arginase-1+, M2 macrophages. Consistent with dectin-1 expression on M2 macrophages, dectin-1 is not required to control parasite burden or induce an inflammatory immune response in vivo. However, dectin-1 deficient mice do exhibit trends towards decreased cyst numbers in the brain and a concomitant increase in AMCase transcripts and AMCase activity ex vivo. Thus, this data suggests that dectin-1 is modulating the immune response and chitinase production during chronic Toxoplasma infection.
Intravital imaging of the colonic mucosa: analysis of iNKT physiology during health and disease (MUC5P.752)
Authors: Zemp Franz , Waterhouse Christopher , Kubes Paul 
Published: January 2023

Abstract
Invariant natural killer T-cells (iNKTs) are specialized innate-like T cells that respond to glycolipid antigens with the rapid secretion of polarizing cytokines. Recent literature has shown that iNKTs are important in mouse models of IBD, but have failed to demonstrate the role of colonic iNKTs specifically. Further, the behavior of these cells has never been observed in the gut. Using intravital spinning disk confocal microscopy and transgenic mice that label iNKT cells, we found that iNKTs actively survey the colonic lamina propria. These cells arrested upon administration of a glycolipid antigen, which was largely presented by colonic epithelial cells. Activation of iNKTs with this ligand produced a rapid inflammatory response typified by proximal colon edema and neutrophil recruitment in the submucosal venules. We extended these studies to the DSS model of colitis, and found iNKT deficiency exacerbates disease. This is even more apparent in CD1d deficient animals, which lack iNKT cells and glycolipid antigen presentation. Current work is evaluating the activation of colonic iNKT cells in this model of colitis. Our data suggest that iNKT activation in the colon results in an acute inflammatory response. However, during colitis, iNKTs and possibly another CD1d-utilizing cell type, play a protective role. Current studies are examining the mechanism(s) behind iNKT activation in the colon and how this activation orchestrates colonic inflammation.
Vitamin A deficiency creates persistent shedding of Citrobacter rodentium and potential reservoirs for the spread of enteric pathogens (MUC5P.755)
Authors: McDaniel Kaitlin , Restori Katherine , Dodds Jeffery , Kennett Mary , Ross A. , Cantorna Margherita 
Published: January 2023

Abstract
Vitamin A deficiency remains a public health concern in developing countries and is associated with increased susceptibility to infectious disease and childhood mortality. We utilized Citrobacter rodentium to model human enteric infections. 40% of vitamin A deficient (A-) mice developed a severe and lethal infection. All of the vitamin A sufficient (A+) mice survived and cleared the infection by day 25. Retinoic acid (RA, active form of vitamin A) treatment of A- mice before and at the time of infection resulted in clearance of C. rodentium with the same kinetics as the A+ mice. Additionally, RA treatment of A- mice at the peak of the infection eliminated C. rodentium within 16 days of treatment. Although the A- mice remained infected at day 37, colon inflammation had resolved to that found in A+ mice. All of the A- and A+ mice were able to clear systemic challenge with C. rodentium indicating that A- mice are able to control systemic bacterial infection. Furthermore, the increased mortality of A- mice during infection was not due to systemic cytokine production or increased C. rodentium pathogenicity. Instead the A- mice seem to have developed a severe gut infection but 60% of the A- mice were able to survive and resolve the inflammation. Our work suggests that improvements in vitamin A status and/or RA treatments may decrease susceptibility to enteric pathogens and eliminate potential carriers from spreading infection to susceptible populations.
Intestine lamina propria CD4+T cells promote the bactericidal activity of macrophages via galectin-9 and Tim-3 interaction during Salmonella typhimurium infection (MUC5P.760)
Authors: Zhang Cai , Zhang Hongru , Zuo Zengyan , Li Zhiyuan , Han Qiuju , Zhang Jian 
Published: January 2023

Abstract
The intestinal immune system is crucial for protection the host from pathogenic infection and maintenance of mucosal homeostasis. The intestine lamina propria macrophages are main effector cells in innate resistance to intracellular microbial pathogens. In the present study, we found that Salmonella typhimurium infection augmented the Tim-3 expression on intestinal lamina propria CD4+T cells and enhanced galectin-9 expression on F4/80+CD11c+macrophages. Moreover, CD4+T cells promote the activation and bactericidal activity of F4/80+CD11c+macrophages via galectin-9 and Tim-3 interaction during Salmonella typhimurium infection. Blockade the interaction of Tim-3 and galectin-9 with a-Lactose significantly attenuated the killing of intracellular bacteria by macrophages. Furthermore, the interaction of Tim-3 and galectin-9 also promoted the formation and activation of inflammasome, which leads to the cleavage of caspase-1 and IL-1β. The secretion of active IL-1β further improved bactericidal activity of macrophages and expression of galectin-9 on macrophages. These results demonstrated the critical role of the cross-talk between CD4+T cell and macrophage, in particularly the interaction of Tim-3 and galectin-9, in antimicrobial immunity and control intestinal pathogen infection.
Effector CD4+CD25- T cells from Stearoyl CoA Desaturase deficient mice drive chronic colitis in RAG-1KO mice (MUC5P.761)
Authors: Yeoh Beng San , Singh Vishal , Vijay-Kumar Matam 
Published: January 2023

Abstract
Fatty acids in lipid bilayer substantially influence membrane fluidity and thus dictate cellular functions. Stearoyl CoA Desaturase (SCD-1) is a lipogenic enzyme that assists in the de novo biosynthesis of oleate (C18:1, n9), a major fatty acid in the phospholipids of lipid bilayer. Accordingly SCD-1KO mice display substantially reduced oleate in cell membranes. We investigated the colitogenic capacity of effector T-cells from SCD-1KO mice and their WT littermates in adoptive T-cell transfer colitis model. Splenic effector T-cells (CD4+CD25-) from age and gender matched WT and SCD-1KO mice were isolated by cell sorting using magnetic beads and flow cytometry. RAG-1KO (n=5) were administered 5.0x105 CD4+CD25- cells/mouse intraperitoneally and monitored for body weights and colitis development. RAG-1KO mice which received CD4+CD25- cells from SCD-1KO mice displayed colomegaly, splenomegaly, elevated colonic, systemic lipocalin-2, a general inflammatory marker and hallmarks of colitis at histologic level. Interestingly, colonic TNFα and IL-10 gene transcripts were significantly reduced in SCD-1KO CD4+CD25- recipients whereas the opposite is true for iNOS and lipocalin-2. Further, systemic sIL-1Ra was significantly reduced in SCD-1KO CD4+CD25- recipients when compared to WT T cell recipients. Our results demonstrate that effector T-cells from oleate poor environment attain colitogenic properties and may be associated with increased immunoreactivity to gut microbiotal antigens.
Supplementation of the aryl hydrocarbon receptor ligand indole-3-carbinol protects mice from Clostridium difficult associated disease. (MUC5P.762)
Authors: Julliard Walker , Fechner John , O'Driscoll Chelsea , Safdar Nasia , Mezrich Joshua 
Published: January 2023

Abstract
We have previously presented the dramatic benefit in survival that the dietary aryl hydrocarbon receptor (AHR) ligand indole-3-carbinol (I3C) confers in C. difficile disease in mice. Other groups have demonstrated that γδ T cells and Tregs play an important role in preventing other models of colitis. The current study sought to determine whether dietary I3C could protect mice from C. difficile by increasing the amounts of protective γδ T cells and Tregs. Furthermore, we examined the role of I3C in preventing antibiotic mediated dysbiosis. Mice were fed an experimental AHR ligand free diet (“base”) or the same diet supplemented with I3C and then infected with C. difficile. Pre-infection, base mice had less γδ T cells and Tregs in their cecum compared to I3C mice. Furthermore, I3C mice had a more robust and less pathologic microbiome pre-infection. Post-infection, I3C mice continued to have higher levels of γδ T cells and Tregs. These data suggest that AHR ligand supplementation can protect the host from C. difficile by reducing antibiotic associated dysbiosis as well as augmenting the local immune system to be more protective from C. difficile associated inflammation. Given the poor treatment options currently available as well as the high morbidity of infection, this represents an exciting new therapeutic possibility.
Differential metabolic features and pathogenic potential of effector CD4 T cells associated with IFNγ production during intestinal inflammation (MUC5P.749)
Authors: Shin Boyoung , Kramer Philip , Darley-Usmar Victor , Harrington Laurie 
Published: January 2023

Abstract
Interferon γ (IFNγ) production is a hallmark feature of mucosal CD4 T cells in inflammatory bowel disease (IBD) patients and multiple mouse models of colitis. However, the contribution of the IFNγ producing cell population to chronic intestinal inflammation remains controversial and requires better understanding. Recent studies have linked cellular metabolism to effector functions of immune cells, but it is unknown if discrete effector T cell subsets present during IBD possess divergent metabolic properties and differential T cell pathogenicity. Using IFNγ reporter mice, viable IFNγ secreting and non-secreting CD4 T cells were isolated from mice with active colitis and examined for metabolic characteristics. While both IFNγ producing and non-producing cells exhibit a high rate of glycolysis, interestingly, the mitochondrial function differs; IFNγ producing cells have lower oxidative phosphorylation and blunted spare respiratory capacity, indicating defective mitochondrial function compared to IFNγ non-producing cells. Consistent with decreased mitochondrial function, IFNγ producing cells have lower cell recovery upon secondary transfer and result in more attenuated intestinal inflammation. Therefore, these data suggest that CD4 T cells that upregulate IFNγ mark a unique effector cell subset with distinctive metabolic features, impaired survival, and reduced pathogenicity, which is paradoxical to the proinflammatory nature of IFNγ during chronic intestinal inflammation.
Non-human primate model of collagen II-induced arthritis: a comprehensive system simulating human rheumatoid arthritis for evaluation of novel therapeutic candidates (THER5P.901)
Authors: Zeng Jia , Dai Feihong , Cheng Yu , Yang Nan , Yang Ping , Zheng Hellen , Fu Yunfeng , Duan Jeff 
Published: January 2023

Abstract
Non-human primate (NHP) model provides the advantage of closer phylogenetic relationship to human and high cross-reactivity to the human target specific biological molecules that do not react with rodent counterpart. PharmaLegacy has been dedicated in developing NHP inflammation models since 2009. Here, we report a NHP model of Collagen II induced Arthritis in Cynomolgus monkey, which developed similar symptoms as human RA in terms of body weight loss, clinical score lifting, joint swelling enlargement and rising PIP incidence. Terminal analysis by radiographic imaging and the multi-dimensional scaling for histopathology and histomorphometry on joints demonstrated similar characteristics of human RA joint deformity, including the significant cell infiltration, loss of proteoglycans (PG) in cartilage, Pannus development, osteophyte formation, loss of articular structure and bone erosion, well supporting for the arthritic disease. Therapeutic treatment with mAb HSF082D significantly reversed the above disease index. In a study of biomarkers profile in Serum or RA joint, CRP/IL-1β/IL-6/IL-17A/RANKL/GM-CSF were found to correlate with the arthritic severity at mRNA or protein levels. Th17 cell activities in synovial fluid were also detected.