bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of maternal silver acetate exposure on immune biomarkers in a rodent model (P1030)
Authors: Babu Uma , Balan Kannan , Bigley Elmer , Pereira Marion , Black Thomas , Olejnik Nicholas , Keltner Zachary , Scott Michael , Sprnado Robert 
Published: January 2023

Abstract
The bacteriocidal effects of silver mixtures are well characterized and as a consequence they are used worldwide in many food contact substances. However, the long-term effect of silver ion exposure on the developing immune system has not been well studied. In this study, 50 d old male and female rats were exposed to 0.4, 4 or 40 mg/Kg body weight silver acetate (AgAc) in drinking water for 10 wk prior to mating and during the 3 week mating period. Sperm positive females remained within their dose groups and were exposed to AgAc during gestation and lactation. Splenic and thymic lymphocyte subsets from postnatal (PN) 4 and 25 d old pups were assessed by flow cytometry for changes in immune biomarker profile. Functional indices included natural killer (NK) activity and mitogen-induced lymphocyte proliferation. Spleens from PN 4 d pups had lower percentages of CD8+ lymphocytes in 4 and 40 AgAc groups; lower CD3 in 40 AgAc group, and reduced Con A response in all 3 AgAc groups. In PN 25 d pups, splenocytes from 40 mg AgAc fed rats had a higher B cell population. However, Con A and LPS mediated lymphocytic responses were significantly reduced in this group. Further, splenic NK activity was higher in PN 25 d old pups exposed to 0.4 AgAc diet. In conclusion, maternal exposure to AgAc had a significant impact on rat immune system development especially in the early lactation period.
SNARE protein requirements for cytotoxic T lymphocyte-mediated cell killing (P1032)
Authors: Giraudo Claudio , Spessott Waldo , McCormick Margaret 
Published: January 2023

Abstract
Cytotoxic T Lymphocytes (CTLs) and Natural Killer (NK) cells selectively kill virally infected or cancer cells by releasing the content of lytic granules at the contact area with target cells called immunological synapse (IS). SNAREs mediate the maturation and exocytosis of “fully-armed” lytic granules at the IS. Deregulation of SNARE-mediated fusion events, such as in Familial Hemophagocytic Lymphohystiocytosis-4 and -5, in which Syntaxin11 and Syntaxin Binding Protein-2 are mutated, respectively; severely impaired lytic granule release and cytotoxicity of CTL and NK-cells. We have identified cognate SNAREs that specifically interact with Syntaxin11 to perform its function during CTL activation and lytic granule release. Biochemical experiments showed that these interacting proteins can form stable SNARE complexes and confirmed the specificity of these interactions. Stimulated Emission Depletion (STED) Superresolution microscopy studies revealed two pools of Syntaxin 11 in CTLs, one localized to a M6PR-containing vesicular compartment, and one localized at the plasma membrane. We have precisely visualized the localization patterns Sytanxin11-interacting partners and other membrane trafficking proteins involved in this process. These results support the conclusion that the identified Syntaxin11-containing SNARE complex plays a crucial role during CTL-mediated cytotoxicity and FHL pathophysiology.
Expression of redox-regulating molecules rather than mitochondrial biogenesis correlate with susceptibility to apoptotic stimuli in human CD62L T cell subsets (P1034)
Authors: Kesarwani Pravin , Al-Khami Amir , Kaur Navtej , Fang Quan , Naga Osama , Beeson Gyda , Johnson Christina , Mayer Elizabeth , Beeson Craig , Nishimura Michael , Mehrotra Shikhar 
Published: January 2023

Abstract
Ex vivo-expanded CD8+ T cells used for adoptive immunotherapy generally acquire an effector memory-like phenotype (TEM cells) and exhibit impaired in vivo persistence and anti-tumor efficacy, compared to T cells with central memory-like phenotype (TCM cells). However, the intrinsic and metabolic factors that attribute to this differential outcome of T cell subsets are not well understood. Here we show that CD62Lhi TCM-like T cells exhibited increased expression of cell surface sulphydryl groups (-SH; thiols), compared to TEM phenotype like CD62Llo TEM-like T cells. This increase in thiols coincided with an up-regulation of the redox-regulating anti-oxidant proteins catalase, SOD1, NRF2 and inversely correlated to the generation of ROS, RNS, extent of T cell proliferation and glycolytic enzymes. Further, we observed that T cells pretreated with rapamycin, a drug that favors the generation of a CD62Lhi TCM-like phenotype of T cells and renders reduced susceptibility to apoptosis, also up-regulated thiol expression and level of anti-oxidative genes. Conversely, thiol donor L-NAC pretreatment reduced mTOR activation. However, both rapamycin and L-NAC treated T cells showed a decrease in oxygen consumption rate, key glycolytic enzymes and expression of PGC1α, a key regulator of mitochondrial biogenesis. These data suggest that modulation of anti-oxidant capacity, mitochondrial metabolism and glycolytic pathway of T cell subsets could increase persistence and improve immunotherapy.
Elucidating immunological synapse organisation: a CAZ-protein study (P1036)
Authors: Maier-Peuschel Monika , Pattu Varsha , Krause Elmar , Rettig Jens 
Published: January 2023

Abstract
The highly adaptive and sophisticated network of our immune system ensures a potent defense against a variety of possible threats. As a part of this machinery, cytotoxic T lymphocytes (CTLs), upon antigen recognition, are inducing programmed cell death by forming an immunological synapse (IS) between CTL and target cell, followed by release of granzyme B and perforin from lytic granules into the synaptic cleft. Whereas some mechanisms underlying the fusion of lytic granules with the plasma membrane have already been determined, temporal and spatial organization of these vesicles right before their fusion at the IS still remains elusive. From chemical synapses in the hippocampus it is known, that vesicle fusion occurs only at the active zone which is being formed by an exclusive number of CAZ (cytomatrix of the active zone) proteins like the 420 kDa protein bassoon. Our qRT-PCR data from primary human and murine CTLs show that CAZ proteins are not only expressed, but are specifically upregulated in activated CTLs, suggesting a possible role during the killing process for example as scaffolding proteins. As specific target killing is a process of paramount importance for survival, it seems likely that vesicle fusion at the IS is precisely organized as well. We present data from primary CTLs derived from CAZ-deficient mice that investigate a possible contribution of CAZ proteins in the organisation and function of the IS.
Protective function of FoxP3+ regulatory T cells in experimental necrotizing enterocolitis (P1010)
Authors: Tran Dat , Dingle Bridgette , Liu Yuying , Fatheree Nicole , Rhoads J Marc 
Published: January 2023

Abstract
Necrotizing enterocolitis (NEC) results from severe intestinal inflammation in premature infants. FoxP3+ regulatory T cells (Tregs) are central to gut homeostasis. While Tregs are reduced in the ileums of premature infants with NEC, it is unknown whether they play a critical function in preventing NEC. This study investigated Treg development in newborn rat pups and their function in regulating experimental NEC induction. Utilizing a rat model of experimental NEC, the ontogeny of T cells and Tregs in newborn pups was characterized by flow cytometry. To investigate the functions of Tregs, newborn pups were given Tregs harvested from adult rats prior to NEC induction to assess clinical improvement and mechanisms of immune regulation. The results revealed that there were few Treg numbers in the terminal ileums of newborn rats and 8-fold reduction after NEC (p=0.002). Adoptive transfer of Tregs significantly improved weight loss, survival from 53% to 88%, and NEC incidence from 87% to 35% (p&lt;0.05). The Tregs suppressed the upregulation of costimulatory molecule CD80 on dendritic cells and inhibited the trafficking of effector cells from leaving the MLN. These findings suggest that there is a developmental delay in Tregs which contributes to the excessive inflammatory state in NEC. Adoptive transfer of Tregs attenuates the induction of NEC by regulating DCs and modulating effector cell trafficking. Treg immunotherapy has a therapeutic potential in the prevention of NEC.
Use of a recombinant miRNA expressing influenza A virus to modulate SOCS1 expression (P6166)
Authors: Stambas John , Jenkins Kristie , Tizard Mark , Izzard Leonard 
Published: January 2023

Abstract
The impact of influenza infection is a major public health concern. Significant morbidity and mortality is associated with infection especially in susceptible populations such as the elderly. Efforts to improve vaccine efficacy and long-term immunological memory are currently areas of significant research interest. miRNAs are naturally occurring 19 to 25 nucleotide (nt) long single stranded RNAs that bind to mRNA and cause inhibition of translation. It is predicted that over 30% of all human genes are targeted by microRNAs (miRNAs), including many immune related genes. For this reason, the artificial use of miRNAs for RNA interference (RNAi) is a rapidly emerging field. However, key limitations exist within the field; most notably delivery of miRNAs to the desired target cell populations in vivo. The suppressors of cytokine signaling (SOCS) proteins are important in the regulation of cytokine production, with SOCS1 and SOCS3 having key roles in innate immunity. miR-155 directly targets SOCS1 and is implicated in the proliferation of various T-cell subsets. This study used a reverse engineered influenza A virus to overexpress miR-155 in infected VERO cells. Using a dual luciferase assay we were able to show a successful reduction in SOCS1 expression. Our long-term goal is to use this viral miRNA delivery system to manipulate the immune system, with the aim of enhancing the efficacy and potency of vaccine induced immune responses.
Generation of memory CD4+ T cells after influenza virus infection requires PSGL-1 and selectin binding activity (P6148)
Authors: Tinoco Roberto , Nguyen Jennifer , Lin Melissa , Bradley Linda 
Published: January 2023

Abstract
CD4+ memory T cells have a critical role in immunity to new influenza viruses. In particular, lung-resident memory CD4+ T cells are essential for production of innate inflammatory cytokines that promote early virus control. However, the contribution of adhesion molecules in regulating CD4+ T cell memory remains unclear. We assessed the role of PSGL-1, a ligand for the selectin family of adhesion molecules that is highly upregulated on CD4+ T cells during infection. In adoptive transfer studies, we found that PSGL-1 KO CD4+ T cells had increased accumulation in the lungs. However, compared to WT cells, few PSGL-1 KO cells persisted into the memory phase and these cells failed to re-expand upon secondary challenge. When PSGL-1 KO mice were re-challenged with a high dose influenza infection, early virus levels in the lungs were increased and KO mice had prolonged morbibity. To address whether functional selectin binding activity was necessary for the development of memory CD4+ T cell responses we used mice deficient in two α-(1,3)-fucosyltransferases, IV and VII (FuT KO) that generate biologically active PSGL-1. FuT KO CD4+ T cells did not display impaired primary responses in the lymph node but showed altered accumulation in the lungs. Like PSGL-1 KO CD4+ T cells, FuT KO cells failed to re-expand upon secondary challenge. Our findings demonstrate a critical role for PSGL-1 and functionally active selectin ligand generation in the development of effective memory CD4+ T cells.
Myeloid derived suppressor cells as therapy for experimental myasthenia gravis (P5229)
Authors: Lin Feng , Li Yan , Qian Shiguan , Fung John , Kusner Linda , Kaminski Henry , Lu Lina 
Published: January 2023

Abstract
We recently demonstrated that hepatic stellate cells (HSC) induce myeloid derived suppressor cells (MDSC) differentiation from myeloid progenitors. In this report, we found that adoptive transfer of the MDSC was effective in reversing disease progression in experimental autoimmune myasthenia gravis (EAMG), a T cell-dependent, and B cell-mediated model for myasthenia gravis (MG). In accordance with ameliorated disease severity, the MDSC-treated EAMG mice showed suppressed AchR-specific T cell responses, decreased levels of serum anti-AChR IgGs and reduced complement activation at the neuromuscular junctions. We also discovered that the MDSC inhibited the proliferation of, and antibody production from B cells activated by anti-IgM or anti-CD40 antibodies in vitro, and inhibited the production of antibodies against a T cell-independent antigen in vivo, indicating that in addition to their T cell inhibitory activities, MDSC also directly inhibit B cells. These results demonstrated that HSC-induced MDSC concurrently suppress T- and B- cell autoimmunity, leading to effective treatment of established EAMG. These data also suggest that the MDSC could be further developed as a novel approach for MG treatment, and even more broadly, for other diseases in which T and B cells are involved in the pathogenesis.
Myeloid resident cells in the islets of Langerhans are key players in presentation of islet antigens and islet trophic function: identification of their phenotype and ontogeny (P5232)
Authors: Calderon Boris , Unanue Emil 
Published: January 2023

Abstract
The entry of autoreactive T cells to their target organ is important in autoimmunity since this initiates the inflammatory process. Islets of Langerhans contain a monocyte/macrophage-derived cell with important features and properties. This islet myeloid cell plays a central role in the interaction with diabetogenic T cells. We have studied in-depth the phenotype of the islet myeloid cells in the NOD and non-diabetic mouse strains and find 2 myeloid subsets: one subset (~85%), expressing MHC-II, CD11c, CD11b, F4/80 and LysM; and a minor subset, (~15%) expressing MHC-II, CD11c, CD11b, CD103, BTLA and LysM. Islet myeloid cells are stable in numbers and decrease 30% 7 days after whole body irradiation. Studies testing eGFP bone marrow transplantation has shown that islet myeloid cells can be replaced as early as 2 weeks post-transplant. Finally, we consider that in addition to their role of antigen presentation, the islet myeloid cell has a different and perhaps a more important role in islet physiology, which is to maintain the health of the islet. When a genetic mutation affects the presence of myeloid cells in islets (CSF-1-deficient mice), islets are reduced in size and show an altered glucose homeostasis. Our findings highlight the importance of the islet myeloid cell as central in presentation of islet antigen and trophic function of the islet. Their phenotype, ontogeny and trophic identification, will bring insights into diagnostics and point of interventions in diabetes.
A novel regulatory pathway for autoimmune disease: binding of partial MHC class II constructs to monocytes reduces CD74 expression and induces both specific and bystander T cell tolerance (P5219)
Authors: Vandenbark Arthur , Meza-Romero Roberto , Benedek Gil , Andrew Shayne , Yu Xiaolin , Mooney Jeffrey , Burrows Gregory 
Published: January 2023

Abstract
Treatment with partial MHC class II-β1α1 constructs (also referred to as recombinant T-cell receptor ligands - RTL) linked to antigenic peptides can induce T-cell tolerance, inhibit recruitment of inflammatory cells and reverse autoimmune diseases. New studies revealed a novel regulatory pathway that involves RTL binding to CD11b+ monocytes through a receptor comprised of MHC class II invariant chain (CD74), cell-surface histones and MHC class II itself for treatment of experimental autoimmune encephalomyelitis (EAE). Binding of RTL constructs with CD74 involved a previously unrecognized DR-α1/CD74 interaction that inhibited CD74 expression on the cell surface and blocked binding and activity of its ligand, macrophage migration inhibitory factor (MIF), resulting in reduced severity of clinical and histological EAE. These findings implicate binding of RTL constructs to CD74 as a key step in both antigen-driven and bystander T-cell tolerance. Furthermore, the natural anti-inflammatory activity and universal expression of the DR-α1 domain supports its possible application as a new therapeutic agent suitable for treatment of a wide variety of human inflammatory conditions.
Small molecule blockers reveal an important role for Orai1-NFAT-orphan receptor pathway in Th17 differentiation (P5169)
Authors: Kim Kyun-Do , Srikanth Sonal , Tan Yossan-Var , Yee Ma-Khin , Jew Marcus , Damoiseaux Robert , Jung Michael E , Shimizu Saki , An Dong Sung , Waschek James A , Ribalet Bernard , Gwack Yousang 
Published: January 2023

Abstract
Orai1 is the pore subunit of the Calcium Release Activated Channels (CRAC) that activate downstream calcineurin-NFAT pathway. Loss of function mutation in Orai1 causes lethality due to severe defects in immune system, which can be rescued by bone marrow transplantation. Using a high throughput chemical library screening, we have identified a class of small molecule blockers that inhibit Orai1. One of the small molecules, compound 5D blocked the activity of a constitutively active mutant of Orai1, suggesting a direct block of ion permeation via Orai1. Primary murine Th17 cells showed highest sensitivity to block by this compound. Treatment with compound 5D not only suppressed cytokine production, but also inhibited differentiation of Th17 cells by suppression of the retinoic-acid-receptor-related orphan receptors RORα and RORγt. Exogenous expression of RORα, RORγt or a constitutive active mutant of NFAT (CA-NFAT) rescued the blocking effect, identifying the Orai1-NFAT- ROR signaling pathway to be important for Th17 differentiation. The defects in cytokine production and differentiation were recapitulated in Th17 cells from Orai1-deficient mice suggesting Orai1 as a specific target of the blockers. In vivo administration of Orai1 blockers effectively delayed the onset and reduced the severity of autoimmune disease in mice. These data indicate that derivatives of compound 5D can be used as chemical templates for development of therapeutic agents to alleviate autoimmune diseases.
A novel CMKLR1 small molecule antagonist suppresses experimental autoimmune demyelinating disease (P5179)
Authors: Zabel Brian , Zhang Jian , Lewen Susanna , Burke Thomas , Carlson Jeffrey , Zoudilova Maria , Sobel Raymond , Butcher Eugene , Graham Kareem 
Published: January 2023

Abstract
Experimental autoimmune encephalomyelitis (EAE) is a widely studied mouse model that shares many clinical and histological features with multiple sclerosis (MS). Chemokine-like receptor-1 (CMKLR1) is a chemoattractant receptor that is expressed by key effector leukocytes in EAE and MS, including macrophages, subsets of dendritic cells, and natural killer cells. We previously showed that clinical and histological EAE is less severe in CMKLR1-deficient (CMKLR1 KO) mice compared with wild-type. In this study, we sought to identify small molecular CMKLR1 inhibitors that recapitulate the CMKLR1 KO phenotype in EAE. We identified α-naphthoyl ethyltrimethylammonium iodide (α-NETA) as a selective CMKLR1 small molecule antagonist that inhibits chemerin-stimulated β-arrestin2 association with CMKLR1, as well as chemerin-triggered CMKLR1+ cell migration. Structure-activity relationship studies indicated that the trimethylamine region is critical for CMKLR1 potency. Prophylactic dosing with α-NETA (i.e. prior to the onset of clinical signs) significantly delayed the onset of EAE induced in C57BL/6 mice by active immunization with myelin oligodendrocyte glycoprotein peptide 35-55. In addition, α-NETA treatment significantly reduced mononuclear cell infiltrates within the CNS. This study provides proof-of-concept data that pharmaceutical targeting of CMKLR1:chemerin interactions may be beneficial in preventing or treating MS.
The RNA virus sensor melanoma differentiation-associated protein 5 mitigates protection from Coxsackievirus B4 and type 1 diabetes in non-obese diabetic mice. (P5211)
Authors: Lincez Pamela , Shanina Iryna , Horwitz Marc 
Published: January 2023

Abstract
Type 1 diabetes is a pancreatic autoimmune disease affecting children and results from both genetic and environmental factors where coxsackievirus B4 (CB4) is the most associated pathogen. Genome-wide association studies have linked variants in MDA5, an RNA virus sensor, with T1D protection. As CB4 is an RNA virus and a likely factor in T1D etiology, we chose to use CB4 to study the role of MDA5 in T1D. CB4 infection enhances T1D in NODs through activation of autoreactive T cells. We hypothesized that MDA5 signals the strength, polarization and regulation of the T cell response after stimulation by agents like CB4 and dampening MDA5 function would alter the T cell response to favour T1D protection. MDA5ko(-/-) were backcrossed to the NOD background and confirmed by SNP analysis. NOD-MDA5ko(10/10) failed to develop spontaneous T1D by 24 weeks when T1D occurred in MDA5het(+/-)(2/10) and wt(+/+)(9/33). While decreased MDA5 expression (confirmed by Western) in MDA5het did not affect spontaneous T1D, it protected MDA5het(4/4 ) from CB4 induced T1D compared to infected wt(2/4). FACS analysis revealed significantly fewer activated effector CD4+ T cells in the spleen and PLNs of MDA5hets. Further, MDA5het PLNs had increased numbers of natural Tregs. These results support our premise that reduced MDA-5 expression alters the T cell response limiting the development of autoreactive effectors and T1D and suggests that dampening MDA-5 function could be a key therapeutic target.
Chemical inhibition of the RORγt-dependent transcriptional network in Th17 cells (P5163)
Authors: Xiao Sheng , Yosef Nir , Kuchroo Vijay 
Published: January 2023

Abstract
RORγt is a master transcription factor required for Th17 development. We identified two novel RORγt-specific inhibitors that suppress Th17 response and Th17-mediated autoimmune disease. To assess their mechanisms of action, we systemically characterize their function within the context of RORγt-directed transcriptional circuitry by integrating RORγt binding data in the presence and absence of drug with corresponding whole-transcriptome sequencing for wildtype and RORγt-deficient cells. RORγt is central in a densely interconnected regulatory network, acting both as a direct activator of genes important for Th17 differentiation and as a direct inhibitor of genes from other T-cell lineages. These inhibitors reversed both of these modes of action, but to varying extents and through distinct mechanisms. While one inhibitor displaced RORγt from its target-loci, the more potent inhibitor affected transcription predominantly without removing DNA-binding. Our work illustrates the power of a system-scale analysis of transcriptional regulation to characterize potential therapeutic compounds that inhibit pathogenic Th17 cells and suppress autoimmunity.