bluuebunny/crossref_metadata_2025_split huggingface dataset:
The novel cytokine IL-17D at the intersection of cellular stress and immune activation (P1364)
Authors: Saddawi-Konefka Robert , O'Sullivan Timothy , Gross Emilie , Tran Miller , Bui Jack 
Published: January 2023

Abstract
Our laboratory has recently identified a novel role for the cytokine IL-17D as an important mediator of the antitumor immune response, and the loss of IL-17D as a key component of tumor progression. Our data show for the first time that overexpression of the cytokine IL-17D is sufficient to induce tumor rejection or growth delay. Tumors expressing IL-17D displayed an increased infiltration of natural killer cells, which gave way to a commensurate increase of M1-type macrophages. While examining the regulation of IL-17D, we came across the surprising finding that IL-17D can be induced by oxidative stress signals via the transcription factor nrf2. Nrf2, responsible for regulating the expression of the anti-oxidant response element genes, is known as the primary cellular responder to oxidative stress. Although the role of nrf2 in cancer biology has been well studied, the role of nrf2 in the regulation of immune responses (especially via IL-17D) has yet to be defined. We find that nrf2 is necessary to induce IL-17D expression in a variety of tumor and non-tumor cells, and that this regulation is active in both tumor progression and viral infection. These studies constitute an intriguing and novel connection between oxidative cellular stress and immune activation.
The development and effector state of innate type-2 lymphocytes is orchestrated by a Gfi1-dependent regulatory network (P1367)
Authors: Lesch Justin , Yan Donghong , Zhou Meijuan , Eastham-Anderson Jeffrey , Diehl Lauri , Lee Wyne , Xu Min , DeVoss Jason , Spoonder Chauncey , Singh Harinder 
Published: January 2023

Abstract
Innate type-2 lymphoid cells (ILC2s) have been shown to play a vital role in protective immune responses against helminthic parasites. The regulatory networks that orchestrate the expansion and effector state of ILC2s are largely unknown. Here we report that the transcriptional repressor Growth factor independence-1 (Gfi1) functions to control the development of ILC2s and the proper specification of their effector state in response to Nippostrongylus brasiliensis infection, papain challenge, and IL33- or IL25 instigated inflammation. Consistent with these findings, IL5 and IL13 levels are reduced and eosinophilia, goblet cell hyperplasia and worm clearance are severely impaired in the peritoneal cavity and lung of Gfi1-/- mice. Analysis of ILC2s reveals a stringent requirement for Gfi1 in their response to IL33 and suggests a prominent role for this signaling pathway in the lung. Strikingly, conditional inactivation of Gfi1 in ILC2s results in reduced expression of IL5 and the mis-expression of IL17. Given that Gfi1 is implicated in reciprocally regulating Th2 and Th17 cell fates, our results reveal it to be a conserved regulatory component which functions in both innate and adaptive immune cells.
A critical role for neddylation in regulating dendritic cell function. (P1369)
Authors: Mathewson Nathan , Toubai Tomomi , Sun Yaping , Tamaki Hiroya , Wang Ying , Oravecz-Wilson Katherine , Kapeles Steven , Hou Guoqing , Reddy Pavan 
Published: January 2023

Abstract
Post-translational modifications (PTMs) of proteins modulate cellular processes. However, the role, specifically that of neddylation (a type of PTM), in regulating immune cells is not known. Neddylation of Cullin-RING ligase (CRL) protein modulates the process of ubiquitination. Thus, CRL activity is critical for the quotidian functions of cells. We explored the role of neddylation in dendritic cells (DCs) utilizing the small molecule inhibitor of neddylation (MLN4924) and the knockdown of SAG, a critical adapter element of CRL. Inhibition of neddylation in the DC significantly reduced the expression and release of several proinflammatory cytokines (TNFα, IL-6) and significantly reduced the ability of DCs to stimulate T cells (P&lt;0.001) following stimulation by several TLR and NLR ligands. This suppression of DC functions was more than that caused by the known anti-inflammatory drug dexamethasone. Mechanistic studies showed that reduction of IκB degradation was critical for the suppression of DCs by the inhibitors of neddylation. This was further confirmed by the absence of NF-κB translocation and function. However, MAPK/ERK pathway was unperturbed by the inhibition of neddylation. Our results demonstrate a novel biological role of inhibition of the PTM, neddylation, in regulating DC functions.
Programmed cell death 4 is an inhibitor of autophagy via targeting ATG5 (P1371)
Authors: Zhang Lining 
Published: January 2023

Abstract
PDCD4 (programmed cell death protein 4), an inhibitor of protein transcription and translation, is involved in tumor and inflammation diseases. However, its underlining mechanisms remain unclear. Autophagy, an evolutionarily conserved catabolic process, is reported to regulate survival and death of cells. Here, we demonstrated that PDCD4 inhibits autophagy in multiple cells including immune cells and tumor cells, by which it controls cell fate. Mechanically, PDCD4 suppressed translation of an essential autophagy related gene ATG5 by its MA-3 domain. Our data indicate that PDCD4 is endogenous inhibitor of Atg5.
Phospholipid scramblase 1 activity mediates phospholipid alterations leading to an endothelial response following hypoxia and reoxygenation (P1295)
Authors: Archer Slone Emily , Fleming Sherry 
Published: January 2023

Abstract
Ischemia, lack of blood flow, and reperfusion, return of blood flow, is a common phenomenon affecting millions of Americans each year. Roughly 30,000 Americans per year experience intestinal ischemia-reperfusion (IR), which is associated with a 70-80% mortality rate. Previous studies of the intestine established a role for innate immune cells and naturally occurring antibodies (Ab) in IR-induced pathology. Furthermore, administration of two monoclonal Ab that recognize phospholipids (PL) or the PL binding protein β2-glycoprotein I (β2-GPI) restores tissue damage in IR-resistant Rag-1-/- mice to wildtype levels. These data indicate involvement of a lipid or lipid-like moiety in mediating IR-induced damage. We hypothesized that PL scramblase 1 (PLSCR1), a protein regulating bilayer asymmetry, is involved in altering the PL of endothelial cells during hypoxia leading to β2-GPI binding and subsequent inflammation. To test our hypothesis, an endothelial cell line, MS1, was subjected to hypoxia (1% O2) followed by reoxygenation as an in vitro model of IR. PLSCR1 transcription, protein expression and activity were assessed. Mass spectrometry detected PL alterations and β2-GPI binding was confirmed by immunohistochemistry. Inflammatory markers including prostaglandins were also evaluated. Our data demonstrate hypoxia and reoxygenation-induced PLSCR1 activity in PL scrambling of endothelial cells. The altered PL arrangement enhances β2-GPI binding and leads to intestinal IR pathology.
Regulation of lytic granules for normal cytotoxic T lymphocyte function (P1140)
Authors: Pattu Varsha , Halimani Mahantappa , Peuschel Monika , Krause Elmar , Rettig Jens 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTLs) function to kill bacterial and viral infected target cells by releasing cytotoxic components such as perforin and granzymes that are contained within lytic granules, into the infected cell. Release occurs via the fusion of lytic granules at the contact zone between infected cell and CTL, the immunological synapse (IS). Defect in the fusion of lytic granules results in the dysfunction of CTLs, the underlying cause of the fatal disease Familial Hemophagocytic Lymphohistiocytosis (FHL). In order for fusion to occur, lytic granules must first arrive, dock and prime at the IS. Mutations within Munc 13-4, a priming factor, for lytic granules, results in FHL subtype-3. We have identified the presence of additional priming factors in CTLs. Targeted gene knockouts of these priming factors resulted in defects in lytic granule fusion at the immunological synapse. These findings demonstrate the presence of more regulatory mechanisms for the most critical event in CTLs, lytic granule fusion. Using high-resolution microscopy and evanescent wave imaging we aim to dissect the precise regulation of lytic granule fusion, which in turn would give us more insight into the regulation and function of CTLs.
Context specific regulation of microRNA targets in the immune system (P1142)
Authors: Loeb Gabriel , Canner David , Khan Aly , Darnell Robert , Leslie Christina , Rudensky Alexander 
Published: January 2023

Abstract
MicroRNAs (miRNAs) are key gene expression regulators—their misexpression can cause both immune dysfunction and cancer. However, most miRNA targets remain unknown, limiting mechanistic insight. Combining genetic and biochemical approaches we generated a transcriptome-wide map of miR-155 targets in T cells. High throughput sequencing of cross-linked immunoprecipitated (HITS-CLIP) Argonaute bound transcript fragments in miR-155 sufficient and deficient cells identified hundreds of direct targets for this miRNA in T cells. This map shed insight on known and novel miRNA phenotypes and demonstrated that miRNAs directly regulate transcripts without canonical target motifs. In addition to T cells, miR-155 is expressed in many innate and adaptive immune cells following activation and its deletion leads to varied cell-type specific phenotypes. We hypothesized that miRNAs have diverse functions in different contexts through regulation of different targets. To reveal the extent and mechanism of context specific miRNA targeting, we employed RNA-seq and HITS-CLIP in wild type and miR-155 deficient T cells, B cells, and dendritic cells. Furthermore, we employed polyA-seq to determine how alternative polyadenylation affects context specific targeting. Immune system complexity is generated from a limited set of genes; by revealing how regulatory factors are repurposed in different cell types, our studies will provide insight into fundamental mechanisms of immunological gene regulation.
NBR1 in the regulation of T cell development and differentiation (P1000)
Authors: Yang Jun-Qi , Liu Hongzhu , Diaz-Meco Maria , Moscat Jorge 
Published: January 2023

Abstract
NBR1 (neighbor of BRCA1 gene 1) is a newly discovered signaling molecule containing a Phox and Bem1 (PB1) domain. PB1-domains are protein interaction modules. PB1 domain-containing molecules participate in diverse biological processes, which also include p62, Par-6, MEK5a, MEKK3, PKCζ and PKC λ/ι besides NBR1. We have recently reported that NBR1 regulates Th2 function and allergic airway inflammation. However, the role of NBR1 in the T cell development and in the regulation of other CD4+ T cell subsets remains to be elucidated. With the use of CD4+ T cell specific conditional NBR1-deficient mice, we found that NBR1 did not affect T cell development. Thymocyte subpopulations (double negative, double positive, CD4 or CD8 single positive cells) were similar between conditional NBR1-deficient mice and their WT littermates. CD4+ and CD8+ cells in the spleen and blood also did not differ significantly between these two groups. Furthermore, NBR1-deficiency did not affect the development of natural Treg in vivo and the differentiation of induced Treg in vitro. However, NBR1 was found selectively regulate Th2 function without affect Th1 cells. Interestingly, our preliminary data indicated that NBR1-deficiency might affect Th17 differentiation in vitro. These data suggest that NBR1 has no effect on T cell development, whereas it may selectively affect some CD4+ T cell subsets, especially Th2 and Th17. Further study is needed to elucidate the underlying mechanisms.
Insulin impairs regulatory T cell function: implications for obesity (P1020)
Authors: Han Jonathan , Patterson Scott , Ehses Jan , Levings Megan 
Published: January 2023

Abstract
Chronic inflammation is known to drive metabolic dysregulation in obesity and type 2 diabetes. Although the precise origin of the unchecked inflammatory responses in obesity is unclear, it is known that over-production of pro-inflammatory cytokines such as TNF-α by innate immune cells has a key role in the development of metabolic dysfunction. One key hallmark of obesity is high levels of the pancreatic hormone insulin, and we hypothesized that there may be an unknown link between hyperinsulinemia and chronic inflammation. Here we show that high levels of insulin impair the ability of regulatory T cells to suppress inflammatory responses via effects on the AKT/mTOR signaling pathway. Insulin strongly activates AKT/mTOR signalling in regulatory T cells, leading to specific inhibition of the production of the anti-inflammatory cytokine IL-10. As a result, insulin hinders the ability of regulatory T cells to suppress the production of TNF-α by macrophages. Regulatory T cells from the visceral adipose tissue of hyperinsulinemic, obese mice also have a decrease in IL-10 production and a parallel increase in production of IFN-γ. These data suggest that the hyperinsulinemia associated with obesity may contribute to the development of obesity-associated inflammation via a previously unknown effect on regulatory T cells function.
PD-1, B7-H1 and IL-10 in protection from kidney ischemia reperfusion injury (P1022)
Authors: Kinsey Gilbert , Jaworska Katarzyna , Ratajczak Joanna , Stevens Brain , Huang Liping , Okusa Mark 
Published: January 2023

Abstract
Regulatory T cells (Tregs) protect the kidney from inflammation, tissue damage and loss of function induced by ischemia-reperfusion (IR) in an IL-10 and PD-1 dependent manner, but the mechanisms of action for Tregs in IR are not completely understood. We hypothesized that Treg PD-1 is engaged by its ligand B7-H1, leading to increased IL-10 production by Tregs, which suppresses inflammation and protects from kidney IR. Similar to our previous observation that WT Tregs lose their ability to protect mice from kidney IR if treated with a PD-1 blocking antibody (ex vivo, prior to adoptive transfer), PD-1 KO Tregs did not offer any protection from kidney IR. In a model of mild renal IR that does not result in significant renal dysfunction in isotype control antibody-treated mice (plasma creatinine (PCr) level: 0.5±0.1, n=5 vs. sham PCr: 0.3±0.1, n=3) blockade of B7-H1 prior to IR causes a marked reduction in renal function (PCr: 1.3±0.3, n=6) accompanied by increased acute tubular necrosis and accumulation of neutrophils in the kidney. Stimulation of isolated WT Tregs with plate-bound anti-CD3, in the presence of IL-2, resulted in significant production of IL-10. In contrast to our hypothesis, addition of plate-bound B7-H1 Ig fusion protein dose-dependently inhibited IL-10 production by WT Tregs. Taken together these results suggest PD-1 and its ligand B7-H1 interact to protect from ischemic kidney injury, but argue against enhanced IL-10 production by Tregs as the mechanism.
Farnesyltransferase inhibitors differentially affect cytokine secretion from CD4+ T cell subsets in mice (P1004)
Authors: Field Kenneth , McMichael James , Dieck Chelsea , Hayes Michael , Gaylo Alison , Crosby Erika 
Published: January 2023

Abstract
Farnesyltransferase inhibitors (FTIs) have been shown to modulate alloreactive immune responses in mouse models of transplantation (A.E. Gaylo, et al., Transplant Immunology 20 (2009) 163-170) and graft versus host disease (A.-K. Hechinger, et al. Haematoligica 98 (2013) 31-40). FTIs can block cytokine secretion from T cells and this activity may explain the drug’s immunomodulatory mechanism. FTIs could modulate immune responses by blocking T cell cytokine secretion thereby affecting the differentiation of stimulated CD4+ T cells into Th1, Th2, or other effector subtypes. To test this hypothesis, we have examined the ability of FTIs to affect cytokine secreation from various T cell subsets and T cell polarization in mice during an allorejection response. Using qPCR we quantified cytokine gene expression in graft-draining lymph nodes (GDLNs) isolated from mice during the rejection of an MHC class II-mismatched skin allograft (bm12 donors to B6 recipients). Using ELISpot assays, we also measured the number of alloreactive Th1 (IFN-γ producing) and Th2 (IL-4-producing) cells in GDLNs. In vitro T cell polarization of polyclonally-stimulated CD4+ T cells isolated from spleen was measured using IFN-γ, IL-4, IL-17, and FoxP3 intracellular staining. We found that FTI treatment differentially affects cytokine secretion from the CD4+ T cell subsets studied. These effects lead to differences in Th1/Th2 cell polarization that could explain the immunomodulatory activity of FTIs.
Intravenous transfer of MOG-pulsed, bone marrow-derived dendritic cells modulates expression of co-stimulatory molecules and cytokines by CD4+ T cells (P5217)
Authors: Zhang Guang-Xian , Zhou Joseph , Ciric Bogolijub , Rostami Abdolmohamad 
Published: January 2023

Abstract
Dendritic cells (DCs) can induce immune tolerance in vivo; however, the mechanisms of DC-mediated tolerance have not been fully elucidated. Here we demonstrate that intravenous (i.v.) transfer of bone marrow-derived DCs pulsed with MOG peptide blocks the development of experimental autoimmune encephalomyelitis (EAE). Transfer of MOG-pulsed DCs leads to down-regulation of IL-17A and IFN-γ production, while the production of IL-10 is up-regulated. The numbers of CD4+IL-10+IFN-γ+ T cells and CD4+FoxP3+IL-10+ regulatory T cells are also increased after MOG-pulsed DC transfer. Intravenous transfer of MOG-pulsed DCs down-regulates protein expression of OX40, CD154 and CD28 on CD4+ T cells, whereas expression of CD152, CD80, PD-1, ICOS and BTLA is up-regulated. Our results indicate that DCs pulsed with MOG inhibit CD4+ T cell activity via modulation of co-stimulatory signaling.
Characterization of a small molecule compound VPR-66 as an inverse agonist for ROR proteins (P5167)
Authors: Lee Hyun-Ku , McAlpine Jennifer , Chitta Sriram , Bouzida Djamal , Singh Gita , Rosenberg Jonathan , Alton Gordon , Singh Sujay 
Published: January 2023

Abstract
Retinoic Acid Receptor (RAR)-Related Orphan Receptors RORα, RORγ and RORγt are key transcription factors required for differentiation of naive CD4+ helper cells into Th17 cells that play an important role in driving autoimmune and inflammatory diseases by secretion of IL-17 and other pro-inflammatory cytokines. Recently development of the Th17 immunomodulators as potential therapeutics for autoimmune diseases has been focused on ROR proteins to regulate differentiation and IL-17 production of Th17 cells. We designed a small molecule compound VPR-66 via the molecular modeling study, which is hypothesized to bind to the ligand-binding domain (LBD) of ROR proteins. VPR-66 was characterized by two different cell-based reporter assays, in which the LBD or full-length of ROR proteins were utilized. In the LBD-based assay, VPR-66 selectively suppressed the RORα and RORγt LBD-mediated transcriptional activity but showed little or no effects on VP16 control protein- or RARα LBD-mediated transcriptional activity. Similarly, in the full-length ROR-based assay, VPR-66 significantly inhibited RORγ- or RORγt-induced IL-17A promoter activity while VPR-66 did not affect the promoter activity in the absence of ROR protein expression. Taken together, it is proposed that VPR-66 acts as an inverse agonist specific for ROR proteins and its binding to ROR proteins interferes with transcriptional complex formation for IL-17 induction.
GABA exerts anti-inflammatory and immunosuppressive effects (P5175)
Authors: Prud'homme Gerald , Glinka Yelena , Wang Qinghua 
Published: January 2023

Abstract
Gamma amino-butyric acid (GABA) is an inhibitory neurotransmitter in the CNS, but it also exerts important functions in the immune system and the islets of Langerhans. Thus, functional GABA receptors have been identified on immune cells and islet cells. Previously, we found that in both autoimmune (NOD) and streptozotocin (STZ)-induced diabetes GABA increases islet-cell mass, while exerting anti-apoptotic effects. GABA induced the regeneration of islet beta cells. It also suppressed inflammatory cytokine production, which is likely important in allowing survival of new islet beta cells. In this study, we examine how GABA exerts immunoprotective effects. We report that GABA suppresses both T cells and macrophages. It increases TGF-beta production and regulatory T cells (Tr or Treg). Notably, GABA inhibits NF-kB activation in both lymphocytes and pancreatic islet beta cells. It also protects beta cells from apoptosis induced by various mechanisms, and this may be related to NF-kB inhibition. The mechanisms by which GABA exerts these effects are largely unknown. However, in immune cells it stimulates GABA type A receptors, which act as ligand-gated chloride channels. This may open voltage-dependent calcium channels in the membrane and decrease intra-cellular calcium levels. Conclusion: GABA acts on both T cells and macrophages and exerts potent anti-inflammatory effects, which are protective in models of type 1 diabetes.
Modulation of immune function occurs within hours of therapy initiation for multiple sclerosis (P5189)
Authors: Sinha Sushmita , Ayers Chris , Mendoza Jason , Cunnusamy Khrishen , Greenberg Benjamin , Frohman Elliot , Karandikar Nitin 
Published: January 2023

Abstract
Daily subcutaneous administration of FDA-approved glatiramer acetate (GA) has beneficial effects on clinical course of relapsing remitting multiple sclerosis (RRMS). Although mechanism of GA-action has been widely investigated and partially understood, immediate immune dynamics following GA-therapy and their significance are unknown. In the present study we characterized immediate effects of GA on phenotype, quantity and function of immune cell subsets in MS patients. PBMC from RRMS patients, either untreated or initiating GA, and healthy donors were obtained at time of admission and subsequently at 4h, 12h, 24h and 72h after first GA-injection. Prominent changes in immune cells were detected within 4-12h post first GA-injection. T-cell modulation included significantly decreased CD4/CD8 ratio, perturbation in homeostasis of predominantly CD8+ T-cells, significant enhancement in CD8+ T-cell mediated suppression and inhibitory potential of induced CD4-suppressors. Changes in APCs were restricted to monocytes and included reduced stimulatory capacity in MLR and significantly increased IL-10 and TNF-α. Our study provides first evidence that GA induced rapid immunologic changes even after first dose. Interestingly, these responses are not restricted to APCs but also include complex modulation of T-cell functionality. Long-term studies will evaluate sustenance of these effects over time and their significance in clinical efficacy of therapy.