bluuebunny/crossref_metadata_2025_split huggingface dataset:
GILT-mediated processing of a self and melanoma antigen in thymic epithelial cells promotes deletion and regulatory T cells (APP3P.113)
Authors: Rausch Matthew , Metzger Todd , Waterfield Michael , Cortez Jessica , Anderson Mark , Hastings Karen 
Published: December 2022

Abstract
Central tolerance is critical to prevent autoimmunity, but limits T cell responses to tumor Ag that are self Ag. We have identified that gamma-interferon-inducible lysosomal thiol reductase (GILT) is required for thymic deletion of CD4+ T cells specific for the self and melanoma Ag, tyrosinase-related protein 1 (TRP1). To define GILT’s role in central tolerance, we show that GILT expression is enriched in thymic APC capable of mediating deletion, medullary thymic epithelial cells (mTEC) and dendritic cells. TRP1 expression is restricted to mTEC. GILT facilitates MHC class II-restricted presentation of endogenous TRP1 by pooled thymic APC. Bone marrow (BM) chimeras demonstrate that GILT expression in TEC is necessary and sufficient for efficient deletion of TRP1-specific thymocytes. In chimeras that express GILT in TEC, TRP1-specific T cells undergo deletion, and chimeras are not protected from melanoma challenge. Although an intermediate level of TRP1-specific T cells develop in chimeras with GILT expression limited to BM-derived cells, only chimeras lacking GILT in both populations are protected from melanoma challenge. Chimeras that express GILT in TECs or in which GILT is limited to BM-derived cells have a substantially higher percentage of TRP1-specific regulatory T (Treg) cells and lower effector:Treg cell ratio. These findings suggest that GILT operates in mTEC to facilitate the presentation of tissue-specific self Ag and promote deletion and development of Treg cells.
Towards the development of artificial antigen-presenting cell (APP3P.116)
Authors: Anikeeva Nadia , Sykulev Yuri , Blanchette Craig , Fischer Nicholas 
Published: December 2022

Abstract
T cell programming into distinct T cell subsets depends in a large extent from the quality and quantity of the signal delivered to T cells by professional antigen presenting cells (APC). However, it is difficult to vary parameters of T cell stimulation using live APC. To calibrate the strength of naïve T cell stimulation, we are developing artificial antigen-presenting cell (APC) that display nanolipoprotein particles (NLPs) presenting pMHC and other ligands incorporated into lipid bilayers built on a glass beads. Soluble analogs of the pMHC assembled on NLPs permits to control the extent of the homotypic ligand clustering and to influence the strength of T cell stimulation. We have found that the apparent equilibrium binding constants of an agonist pMHC/NiNLPs were strongly dependent on pMHC density. A higher density of pMHC on NiNLPs resulted in a much stronger response even though the concentration of the pMHC/NiNLPs at the lower pMHC density was 10-fold higher. We have also demonstrated that bilayers on glass beads were fluid allowing the mobility of pMHC/NLPs on the surface of engineered APC. In a model experiment, T cells concentrated the ICAM-1/pMHC/NLPs conjugates at the contact surface indicating that these conjugates were able to move on the bilayers mimicking direction stimulation of the T cells with live APC. These and other data will be discussed to demonstrate the utility of engineered APC for stimulation of naïve T cells ex vivo.
Neuropsychiatric lupus occurs independently of systemic autoimmunity (BA6P.120)
Authors: Stock Ariel , Wen Jing , Doerner Jessica , Putterman Chaim 
Published: December 2022

Abstract
Neuropsychiatric disease is a common manifestation of systemic lupus erythematosus (SLE). Potential mechanisms underlying neuropsychiatric SLE (NPSLE) include thrombosis, complement deposition, brain-reactive autoantibodies, and cytokine mediated inflammation. However, it is not clear whether the neuropsychiatric manifestations of SLE are secondary to systemic disease, or a primary pathogenic process in the brain. In order to distinguish between central nervous system (CNS) and hematopoietic contributions, we generated bone marrow chimeras between lupus-prone MRL/lpr and congenic control MRL/+ mice. We monitored systemic disease progression, performed extensive behavioral testing, and analyzed brain tissue for evidence of inflammation. MRL/lpr (donor) → MRL/lpr (recipient) chimeric mice showed significantly elevated autoantibodies and marked proteinuria while MRL/+→MRL/+ and MRL/+→MRL/lpr did not, indicating abrogation of the MRL/lpr systemic disease phenotype. Surprisingly, MRL/+→MRL/lpr mice displayed a behavioral phenotype consistent with MRL/lpr→MRL/lpr mice, including depression-like behavior and increased spatial memory deficits. In contrast, MRL/+→MRL/+ mice behaved normally. Additionally, there was elevated RANTES expression, cellular infiltration, and neurodegeneration in MRL/lpr→MRL/lpr and MRL/+→MRL/lpr chimeric mice. The data presented herein indicate that the MRL/lpr CNS is sufficient to mediate NPSLE development absent significant hematopoietic contribution.
33D1+ dendritic cells in the maintenance of Il-2 dependent regulatory T cells (BA6P.130)
Authors: Stolley Michael 
Published: December 2022

Abstract
Regulatory T cells (Tr cells) are potent anti-inflammatory cells that restrain autoimmune and inflammatory disease by dampening immune responses to self- and foreign-antigens. Given their potent immunosuppressive properties, manipulation of the Tr cell compartment is an attractive therapeutic strategy to either boost or inhibit immune responses in a variety of clinical settings. Proper implementation of Tr cell based immunotherapies necessitates a clearer understanding of how these cells are maintained in different anatomical niches under homeostatic conditions. Although it has been recognized for several years that dendritic cells (DC) help control Tr cell abundance and function, the mechanisms of this cross-talk, and a more specific definition of the cellular and molecular components underlying Tr cell maintenance by DCs, has not been elucidated. Here we show Il-2 signaling in Tr cells, which is critical for their proper homeostasis and function, is DC dependent. Intriguingly, CD8α+ DCs are not required for Il-2 activation in Tr cells and their frequencies are in fact elevated in BATf3-/- mice lacking CD8α+ DCs. Instead 33D1+ DCs have a critical yet indirect function in the generation of Tr cell supportive Il-2, and this effect is largely mediated by the micro-anatomical positioning of these cells within T cell zones of secondary lymphoid organs.
Effector T cell resistance to suppression by regulatory T cells as a mechanism of SLE-accelerated atherosclerosis (BA6P.132)
Authors: Wilhelm Ashley , Rhoads Jillian , Moore Jared , Major Amy 
Published: December 2022

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by T and B cell activation and circulating autoantibodies. Common early causes of mortality include kidney disease and infection, while there is a striking increase in atherosclerosis risk for those surviving more than a year after diagnosis. Despite identification of this increased risk more than 40 years ago, mechanisms for SLE-accelerated atherosclerosis remain elusive. The B6.Sle1.2.3 (B6.SLE) mouse model of SLE contains three genetic loci that confer SLE susceptibility. Transplantation of bone marrow from B6.Sle1.2.3 into LDLr-/- mice results in increased atherosclerosis accompanied by more CD4+ T cells in the lesions, pointing to T cell dysregulation as a contributing factor. We confirmed a central role for CD4+ T cells in this process by demonstrating that transfer of B6.SLE CD4+ T cells into Rag-/-, LDLr-/- mice resulted in increased atherosclerosis compared to B6 CD4+ T cells. In the current study, we hypothesized that acceleration of atherosclerosis in SLE is due to resistance of B6.SLE CD4+ effector T cells (Teff) to suppression by regulatory T cells (Treg). Our data show that, although B6.SLE Treg have normal suppressive capacity, B6.SLE Teff are resistant to this suppression. In co-transfer experiments, results suggested an IL-17-mediated mechanism, by which B6.SLE Teff elicit increased IL-17 production by B6 Treg, much like levels produced by B6.SLE Treg.
NLRP12 negatively regulates autoinflammatory disease by modulating T cell responses and IL-4 production (BA6P.133)
Authors: Lukens John , Gurung Prajwal , Kanneganti Thirumala-Devi 
Published: December 2022

Abstract
Missense mutations in the Nod-like receptor Nlrp12 are associated with periodic fever syndromes and atopic dermatitis in humans, however the cellular and molecular mechanisms involved in NLRP12-mediated regulation of immune responses remain poorly defined. In this study, we focused on identifying the roles of NLRP12 in shaping inflammatory T cell responses and T cell-mediated disease. We found that immunization of NLRP12-deficient mice with myelin antigen to induce experimental autoimmune encephalomyelitis (EAE) provoked the generation of hyperinflammatory T cell responses in the periphery. Surprisingly, NLRP12-deficiency did not cause exacerbated forms of paralyzing EAE disease, instead Nlrp12-/- mice developed atypical neuroinflammatory symptoms that were characterized by ataxia and loss of balance. Non-classical EAE pathogenesis was characterized by altered regulation of canonical and non-canonical NF-kB signaling in the CNS. Furthermore, enhanced IL-4 production was identified to promote the development of atypical EAE disease in Nlrp12-/- mice. Interestingly, Nlrp12 was found to be highly expressed in T cells and purified NLRP12-deficient T cells exhibited hyperproliferation and enhanced NF-kB signaling in response to TCR stimulation. These results define an unexpected role for NLRP12 as an intrinsic negative regulator of T cell-mediated immunity, and identify altered NF-kB regulation and IL-4 production as key mediators of NLRP12-associated disease.
Fn14 deficiency protects lupus-prone mice from cutaneous lesions induced by UVB irradiation (BA6P.135)
Authors: Doerner Jessica , Friedman Adam , Burkly Linda , Putterman Chaim 
Published: December 2022

Abstract
Cutaneous lupus erythematosus (CLE) in one of its various forms is seen in the majority of patients with systemic lupus. Photosensitivity is common in lupus, and sunlight (via UVB irradiation) is a well-recognized trigger of CLE. A TNF superfamily member, TNF-like weak inducer of apoptosis (TWEAK), is a soluble cytokine that binds to its sole known signaling receptor, Fn14. TWEAK and Fn14 are upregulated with inflammation and/or injury, and this pathway is important in critical biologic processes including angiogenesis, cell survival, and local inflammation. Previously, we found that lupus-prone MRL/lpr mice deficient for Fn14 (Fn14 KO) had markedly attenuated skin disease (which develops spontaneously in this strain) as compared to Fn14 wild-type (WT) mice. Here, we demonstrate that absence of the Fn14 receptor is protective against the expected UVB-induced eruption. MRL/lpr Fn14 WT mice developed significantly more severe skin involvement following exposure to UVB when compared to strain, age, and gender matched Fn14 KO mice. Histologically, MRL/lpr Fn14 WT mice had significantly more apoptotic keratinocytes, basement membrane degeneration, and dermal inflammation comprised of macrophages following UVB. RT-PCR revealed higher levels of CXCL9, CXCL10, CXCL11 and RANTES in MRL/lpr Fn14 WT mice. Our data suggests a novel role for the TWEAK/Fn14 signaling pathway in the induction of cutaneous lupus lesions following UVB.
Exposure to inhaled atmospheric particulate matter enhances autoimmunity. (BA6P.125)
Authors: Fechner John , Julliard Walker , O'Driscoll Chelsea , Mezrich Joshua 
Published: December 2022

Abstract
Epidemiological studies have demonstrated a relationship between exposure to atmospheric particulate matter (PM) and autoimmune disease. Multiple mechanisms have been proposed, including our research showing PM exposure in vitro can enhance Th17 differentiation in an aryl hydrocarbon receptor-dependent manner. The current study sought to determine the impact of PM exposure in vivo on immunity outside of the lung. In the 1st experiment, male B6 mice (n = 4 per group) were treated every 3 days with either vehicle or 800ug of SRM1949b, a representative sample of urban dust particles, in 20ul PBS i.n. for a total of 5 doses. One day after the last dose, splenocytes were cultured for 4 days with anti-CD3 and cytokines were measured. Treatment with PM increased IL-17A 3-fold but had no impact on IFNγ expression. In the 2nd experiment, female B6 mice (n = 4 per group) were treated every 3 days with vehicle or PM for a total of 9 doses. On the day of the 5th dose, a standard EAE protocol commenced. Clinical scoring using a 5-point scale began on d7 and ended on d21. All mice in both groups displayed EAE symptoms. Mice treated with PM had significantly higher mean peak score (3.5 + 0.3 vs 2.5 + 0.3 for PBS), trended toward having a higher mean clinical score and had earlier onset of disease. Together, these data suggest that inhaled PM exposure can impact immune responses beyond the lung, and support a mechanism where inhaled pollution aggravates autoimmunity.
Mast cells play a key role in autoinflammatory disease development in PSTPIP2-deficient cmo mice (BA7P.141)
Authors: Craig Andrew , Lee J.H. , Sharma Namit , Chitu Violeta , Stanley E. 
Published: December 2022

Abstract
Chronic recurrent multifocal osteomyelitis (CRMO) is an autoinflammatory disease characterized by fevers, bone lesions, and skin lesions. A CRMO-like disease develops in cmo mice, which harbor a mutation in proline serine threonine phosphatase-interacting protein 2 (PSTPIP2). Loss of PSTPIP2 in cmo mice has been linked to elevated macrophage and osteoclast activation, and IL-1β-dependent disease progression. Since mast cells were detected in cmo lesions, we have tested the role of mast cells in cmo disease. In diseased cmo mice, increased connective tissue mast cells were observed in both normal skin and within skin lesions. Crossing cmo mice with a transgenic model of mast cell deficiency (Mcpt5-Cre:Rosa26-Stopfl/fl-DTa) resulted in mice (cmo/MC-) that were protected from developing cmo disease, including skin lesions and joint swelling. Fewer osteolytic lesions were also observed in cmo/MC- mice by micro-computed tomography. Within these tissues, cmo/MC- mice had reduced levels of IL-1β, and less activated immune cells in popliteal lymph nodes compared to cmo. Intravital microscopy experiments on young cmo mice revealed increased leukocyte recruitment in response to lipopolysaccharide compared to cmo/MC- and wild-type mice. Together, these results implicate mast cells in promoting leukocyte recruitment to inflamed tissues during autoinflammatory disease progession, and suggest that therapies targeting mast cells or their mediators may benefit CRMO patients.
Immune complex-induced, nitric oxide-mediated vascular endothelial cell death by phagocytes (BA7P.159)
Authors: Shashidharamurthy Rangaiah 
Published: December 2022

Abstract
Immune-complex (IC) mediated vascular endothelial cell death linked to the release of toxic free radicals by activated inflammatory cells. In particular, nitric oxide (NO), a highly reactive free radical, is implicated in many IC-mediated inflammatory disorders. However, the mechanisms of NO-induced endothelial cells death during autoimmune vasculitis have not been clearly understood. Therefore, we investigated the molecular mechanisms of extracellularly secreted NO by IC-activated phagocytes on endothelial cells using in-vitro and in-vivo vasculitis models. When co-incubation of antibody-coated endothelial cells, murine macrophages released significantly high levels of NO (12 folds) into the culture media. In addition exogenously added NO induced upregulation of pro-apoptotic genes such as Bax (15 fold), Bak (35 fold), cytochrome-C (11 fold) and caspase-3 (30 fold) in endothelial cells. Further, in vivo studies revealed that circulating ICs deposit and lead to accumulation of inflammatory cells in the capillaries but not in large arteries. In addition, the IC-mediated damages to vascular endothelium are prevented by uncoupling IC and inflammatory cells interaction. In summary these results suggest that, though the complexity of NO-induced cell death signaling exists due to its differential (dichotomous) cellular activities, it is absolutely dangerous for cell survival at sustained and chronic level of production as possibly occurring in IC-mediated autoimmune vasculitis
Heat shock protein 90, a potential biomarker for type I diabetes, is secreted by human pancreatic beta cells in response to cytokine stress (BA7P.161)
Authors: Gardiner Gail , Guindon Lynette , Perez Liliana , Evans-Molina Carmella , Blum Janice 
Published: December 2022

Abstract
Heat shock protein 90 (HSP90) is a molecular chaperone that regulates diverse cellular processes by facilitating the activities of a wide variety of protein clients. Recent collaborative studies from our lab have shown that serum HSP90 levels are elevated in newly diagnosed type I diabetic patients, thus distinguishing HSP90 as a potential biomarker for this autoimmune disease. The clinical onset of type 1 diabetes (T1D) is known to be associated with insulitis, the infiltration of inflammatory cells into the pancreatic islets. Therefore, to test the hypothesis that inflammatory stress results in HSP90 release from pancreatic islets, we treated primary human cadaveric islets from non-diabetic donors with a cocktail of inflammatory cytokines IFN-γ, TNF-α, and IL-1β and found that increased levels of HSP90 are secreted by cytokine-treated islets. Flow cytometry of dispersed pancreatic islets suggests beta cells may be responsible for the release of this chaperone in response to cytokine treatment. Further studies with cytokine-treated human beta cell lines βLOX5 and 1.1B4 support this finding. In contrast, beta cell lines do not secrete HSP90 upon treatment with thapsigargin, suggesting HSP90 is not released in response to beta cell endoplasmic reticulum stress, which is also known to precede the onset on T1D. Studies are underway to determine the exact mechanism by which HSP90 is released in response to cytokine stress as well as to elucidate the role of HSP90 in T1D etiology.
The role of neutrophils in B cell dysregulation in systemic lupus erythematosus (BA7P.146)
Authors: Bird Anna , Rangel-Moreno Javier , Anolik Jennifer 
Published: December 2022

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease involving dysregulation of both innate and adaptive immune compartments. The hallmark of SLE is loss of B cell tolerance and production of auto-antibodies, which results in tissue damage and inflammation. Factors driving loss of B cell tolerance are incompletely characterized, but include failure of central tolerance mechanisms, inadequate clearance of cell debris, and alterations in the cytokine milieu, including elevated interferon alpha (IFNα). Our lab and others have shown that SLE neutrophils are a significant source of IFNα as well as self-antigen in the form of apoptotic debris and neutrophil extracellular traps (NETs). However, it is unknown whether neutrophils contribute to B cell activation and tolerance loss in lupus. To address this, we examined both splenic and bone marrow (BM) tissues from lupus prone mice to determine whether neutrophils secrete cytokines that alter the inflammatory milieu. Our data shows that neutrophil numbers and apoptosis are increased in lupus BM. BM neutrophils from lupus prone mice have elevated production of the B cell survival cytokines BAFF and APRIL, as well as IFNα. There was an enrichment of IFNα-producing neutrophils in the spleen in close proximity to both B and T cells. These data suggests that neutrophils likely participate in the creation of an inflammatory microenvironment that sustains auto-reactive B cells and exacerbates SLE pathogenesis in the spleen and the BM.
Induction of autoimmune disease by adoptive transfer of an atypical NK cell subset (BA7P.147)
Authors: Bolland Silvia , Voynova Elisaveta , Skinner Jeffrey , Scott Bethany , Qi Chen-Feng 
Published: December 2022

Abstract
Several mouse models of SLE, including FcgRIIB-KO and TLR7tg mice, develop an expansion of an atypical NK cell subset with functional similarity to cells referred as IKDCs or pre-mNKs in other systems and identified as NK1.1+CD11c+CD122+MHC-II+. These cells belong to the NK cell lineage because they depend on IL15 and express E4BP4. Functionally they are cytotoxic, produce type I and type II interferons upon activation and they are efficient antigen presenting cells both through MHC-II expression and in cross-presentation to CD8s. These atypical NK cells are responsive to TLR stimulation and thus are most abundant in mice with high copy number of the Tlr7 gene. They are highly proliferative as assessed by in vivo BrdU incorporation. Transferring 4 million atypical NKs purified from spleens of SLE-prone mice into WT induces a 2-week-long wave of inflammatory cytokines in the serum, a sustained increase in T cell activation and follicular helper cells for the following months, and a progressive expansion of dendritic cells, monocytes and granulocytes. Furthermore IL15 deficiency, which impedes development of NK cells, ameliorates the autoimmune pathology of TLR7tg mice. These results suggest that cells of the NK lineage can develop into cytokine producing/antigen-presenting cells that affect the priming and progression of systemic autoimmune disease.
CD4+ T cells display increased cross reactivity when primed with peptides of lower affinity for MHC (BA8P.165)
Authors: Blanchfield Jennifer , Evavold Brian 
Published: December 2022

Abstract
Studies propose the degree of T cell cross reactivity for peptide antigens is associated with similarities in the amino acid residues contacting TCR. Here we advance this concept by assessing how the stability of peptide for MHC contributes to cross reactivity in demyelinating autoimmune disease by evaluating two CNS antigens MOG35-55 (myelin oligodendrocyte glycoprotein) and NFM15-35 (neurofilament medium protein). These peptides share identical TCR contact residues yet vary at 3 MHC anchors such that MOG exhibits greater stability than NFM for I-Ab. This difference dictates encephalitogenic potential as well because MOG promotes demyelination and NFM does not. Polyclonal CD4+ T cells primed with MOG or NFM were probed for affinity and cross reactivity using MHC monomers presenting the priming peptides or the encephalitogenic peptides MOG(42P) and MOG(47A), which vary from wild type MOG at TCR contact residues. The 2D micropipette adhesion frequency assay used to assess these parameters showed that NFM and MOG expanded splenic CD4+ T cells that recognized 47A and 42P with similar frequencies and low affinities ~1.8 x10-6 μm4. Interestingly though, NFM specific cells had a greater capacity to recognize multiple peptides than MOG specific cells and this potential was unique to individual clones. Thus, peptide stability for MHC influences the degree of cross reactivity in peripheral tissues and provides a unique point of comparison for studying cross reactivity in the CNS.