bluuebunny/crossref_metadata_2025_split huggingface dataset:
A specific tolerance inducing vector for therapeutic treatment of autoimmune diseases (THER5P.842)
Authors: Hansson Charlotta , Schön Karin , Hasselberg Annemarie , Lycke Nils 
Published: January 2023

Abstract
We have developed a novel platform for intranasal treatment of autoimmune diseases in which ADP-ribosylation determines whether immunity or tolerance is induced. Hence, cholera toxin A1-subunit based immunomodulation through the CTA1-peptide-DD fusion protein promotes enhancement, while inactive mutants induce suppression. Whereas the target population after intranasal administration of both constructs was CD103+CD11blow DC’s, both active and inactive mutants induced strong CD4 T cell priming, but they differentially affected CD4 T cell differentiation. This way inactive mutant constructs generated regulatory CD4 T cells producing IL-10 and effectively preventing experimental autoimmune encephalitis (EAE) and collagen-induced arthritis (CIA), when carrying relevant peptides. Targeted DCs expressed low levels of CD80, CD86 and CD40, while, by contrast, ADP-ribosylating CTA1-peptide-DD constructs gave significantly enhanced co-stimulation. Suppression was global in that regulatory T cells following treatment were able to suppress adoptively transferred naïve CD4 T cells and could subsequently maintain tolerance. The use of specifically tolerance-inducing fusion proteins may be a way forward in the search of effective treatments against autoimmune diseases. The immunomodulating effect on the CD103+CD11blow DCs will be described and the requirement for CTA1-binding to Gsa evaluated in detail, using Cre-lox Gsa-deficient mice.
Conjugated linoleic acid reduces inflammation via PPARγ in murine autoimmune glomerulonephritis (THER6P.861)
Authors: Hammond Sarah , Regna Nicole , Chafin Cristen , Reilly Christopher 
Published: January 2023

Abstract
Conjugated linoleic acid (CLA) has been shown to reduce inflammation via Peroxisome Proliferator-Activated Receptor (PPAR)-γ in inflammatory disorders. We sought to determine whether CLA isomers would reduce inflammation via PPARγ in cultured mesangial cells, and in murine autoimmune glomerulonephritis. Mesangial cells were cultured with pure CLA isomers (c9,t11 or t10,c12) or a 50:50 mixture prior to immune stimulation. Next, cultured mesangial cells were transfected with siRNA targeting PPARγ and treated with CLA isomers prior to immune stimulation. ELISA and RT-qPCR were performed to measure cytokine production and mRNA expression. Treatment with c9,t11 CLA reduced IL-6 production in cultured mesangial cells, but not in siRNA-treated mesangial cells, suggesting c9,t11 CLA acts through PPARγ. IL-1RA mRNA expression was significantly higher in c9,t11-treated siRNA-transfected cells than in t10,c12-treated cells, suggesting alternate pathways in addition to PPARγ. For in vivo studies, wild-type and PPARγ -/- mice were fed a control diet or a CLA-supplemented diet (50:50 mixture of c9,t11 and t10,c12) prior to induction of glomerulonephritis by anti-glomerular basement membrane antisera administration. CLA treatment reduced renal TGFβ in the CLA-treated wildtype mice, but not in the CLA-treated PPARγ-knockout mice. Taken together, these studies suggest that CLA acts in part through PPARγ to decrease inflammatory mediator production in autoimmune glomerulonephritis.
A pharmacological protein kinase D inhibitor ameliorates experimental arthritis. (THER6P.850)
Authors: Yi Ae-Kyung , Yoon Tae Won , Brand David , Kim Young-In , Park Jeoung-Eun , Stuart John 
Published: January 2023

Abstract
While the factors that initiate and perpetuate inflammation in rheumatoid arthritis (RA) are not well understood, it is thought that RA may be initiated in part by signaling through Toll-like receptors (TLRs). Disruption of the TLR signaling pathway may provide an opportunity to halt the RA process in the earliest stages of development. We previously found that protein kinase D1 (PKD1) is essential for proinflammatory responses mediated by MyD88-dependent TLRs. Here, we investigated whether PKD1 is involved in exaggerated proinflammatory responses seen in RA patients, and in the development of experimental arthritis. PKD1 was constitutively hyperactivated in synoviocytes of RA patients, and was rapidly activated in synoviocytes of normal donors and RA patients in response to TLR/IL-1R ligands. Silencing PKD1 expression in RA synoviocytes resulted in substantial inhibition of both spontaneous and TLR/IL-1R-mediated expression of cytokines/chemokines. Suppression of PKD1 expression also resulted in inhibition of TLR/IL-1R-mediated cytokine production in IL-1R antagonist-deficient (IL-1rn-/-) macrophages. In addition, daily treatment with a PKD inhibitor substantially reduced the incidence and severity of spontaneously occurring inflammatory arthritis in IL-1rn-/- mice, and collagen-induced arthritis in humanized HLA-DR1 mice. This implies that PKD1 might be one of the key regulatory factors that modulate proinflammatory responses in RA, and can be a therapeutic target for RA.
IL-12/Th1 and IL-23/Th17 biliary microenvironment in primary biliary cirrhosis (THER6P.852)
Authors: Yang Chen-Yen , Ma Xiong , Tsuneyama Koichi , Huang Shanshan , Takahashi Toru , Chalasani Naga , Bowlus Christopher , Yang Guo-Xiang , Leung Patrick , Ansari Aftab , Wu Linda , Coppel Ross , Gershwin Merrill 
Published: January 2023

Abstract
The interleukin (IL)-12/IL-23 mediated Th1/Th17 signaling pathway has been associated with the etiopathogenesis of primary biliary cirrhosis (PBC). To address the cytokine microenvironment specifically in the liver, we examined the localized expression of IL-12/IL-23 molecules and their corresponding receptors by immunohistochemistry with an extensive panel of antibodies with specificity for IL-12p70/p35, IFN-γ, IL-12RB2, IL-23p40/p19, IL-17 and IL-23R using liver from PBC (n=51) and non-PBC (n=80) control liver disease patients. Sections of portal tracts were blindly evaluated and individually scored. We report herein that although IL-12/Th1 and IL-23/Th17 staining were detected in all of the liver sections, they were primarily localized around the damaged interlobular bile ducts in PBC. Most importantly, the Th1/Th17 balance skewed toward Th17 prominently in liver tissues from advanced PBC patients with intensive secretion of IL-23p19 by inflamed hepatocytes around IL-23R, IL-12RB2, and IFN-γ expressing degenerated cholangiocytes. In conclusion, our novel finding on the direct association of Th17 skewing and disease severity illustrates the significance of the IL-23/Th17 pathway in the perpetuation of IL-12/Th1-mediated immunopathology in PBC and has important therapeutic implications. The imbalance toward Th17 at advanced stages of PBC suggests the necessity to consider blockade of both IL-12/Th1 and IL-23/Th17 signaling for therapeutic intervention.
Therapeutic treatment of multi-organ system, obstructive pulmonary and scleradermatous chronic graft-versus-host disease with the BTK and ITK inhibitor Ibrutinib (TRAN3P.873)
Authors: Flynn Ryan , Dubovsky Jason , Harrington Bonnie , Reichenbach Dawn , Yiming Zhong , Yang Carrie , Towns Will , Lehman Amy , Johnson Amy , Muthusamy Natarajan , Devine Steven , Jaglowski Samantha , Serody Jonathan , Murphy William , Munn David , Luznik Leo , Maillard Ivan , Koreth John , Cutler Corey , Soiffer Robert , Antin Joseph , Ritz Jerome , Panoskaltsis-Mortari Angela , Byrd John , Blazar Bruce 
Published: January 2023

Abstract
Chronic GVHD (cGVHD) is dependent on the ability of donor bone marrow (BM) B cells to produce pathogenic antibody (Ab) deposited in target organs, which is associated with increased germinal centers (GCs). BTK is necessary for B cells to enter GCs and ITK is necessary for T cell activation. We hypothesized that alloreactive GC B cells and T follicular helper cells require BTK or ITK and inhibition by ibrutinib (Ib) would prevent cGVHD. We evaluated the therapeutic effect of Ib in two models of cGVHD: a MHC-disparate (B6→B10.BR) multi-organ model complicated by bronchiolitis obliterans (BO) and a sclerodermatous (SCL) minor-mismatch (LP/J→B6) model. Mice treated with Ib had a decrease in pathogenic Ab, collagen deposition and GC B cells in target organs similar to healthy control mice. In contrast to cGVHD mice, pulmonary function in Ib treated mice was similar to control mice. To confirm that BTK and ITK are required for cGVHD we used donor mice with an ablation of BTK or ITK. Absence of BTK in B cells impeded GCs, Ab deposition and BO. Mice transplanted with T cells lacking ITK and WT BM did not develop BO. In the SCL model, mice treated with Ib did not develop clinical signs of cGVHD including less alopecia and SCL lesions. Furthermore, there was a decrease in lymphocyte infiltration surrounding the bronchioles and nephrons in treated mice. The inhibition of TEC-family kinases with Ib is a novel therapeutic for multi-organ system cGVHD with BO or SCL manifestations.
Endogenous memory CD8 T cells exhibit increased early proliferation in cardiac allografts subjected to prolonged ischemia (TRAN3P.880)
Authors: Su Charles , Iida Shoichi , Abe Toyofumi , Fairchild Robert 
Published: January 2023

Abstract
Memory T cells induced to microbial infections provide protection to subsequent exposures but can have cross-reactivity with allogeneic MHC molecules. High numbers of memory T cells with donor reactivity in human and non-human primate recipients prior to transplant are associated with poorer allograft survival and resistance to costimulatory blockade strategies given to induce long-term graft survival. Rodent transplant models have failed to replicate the detrimental impact of endogenous memory T cells on allograft survival and tolerance induction, raising questions about the sufficiency of these models to study this component of alloimmunity. We have recently shown a direct association between increasing duration of cold ischemic storage and numbers of early graft infiltrating memory CD8 T cells that markedly up-regulate effector functions and directly mediate costimulatory blockade resistant rejection of cardiac allografts in a mouse model. Here we show that endogenous memory CD8 T cells exhibit increased early proliferation in grafts subjected to prolonged ischemia that is supported by memory CD4 T cells. Recipient depletion of CD4 T cells or treatment with MR1 (anti-CD40L) at transplant significantly decreased early memory CD8 T cell proliferation and extended survival of grafts subjected to prolonged cold ischemia. These studies provide novel insights into endogenous memory CD8 T cell mediated graft injury and the poor outcomes associated with cadaver donor grafts.
Defining the alloreactive T cell repertoire using high-throughput sequencing of mixed lymphocyte culture (TRAN3P.884)
Authors: Emerson Ryan , Matthew James , Robins Harlan , Leventhal Joseph 
Published: January 2023

Abstract
The cellular immune response is a major barrier to patient tolerance of allografts. Positive followed by negative selection in the thymus ensures that T cell populations have some binding affinity for self-HLA, but culls T cells with high specificity for peptides presented on self-HLA. However, in the context of a patient with an allograft, these processes also allow for mature T cells with off-target specificity for self-peptides presented on allo-HLA. Currently, the breadth of the alloreactive T cell repertoire is unknown, as well as whether the alloreactive T cell repertoire is stable over time. We have used high-throughput sequencing to characterize the alloreactive T cell repertoire in three pairs of healthy adults using mixed lymphocyte culture, analyzing 4 reactions per pair: duplicate MLR reactions at baseline followed by duplicate MLR reactions 3 months later. Our results indicate that thousands of T cell clones proliferate in mixed lymphocyte culture, suggesting a very broad alloreactive T cell repertoire. This repertoire is consistent across biological replicates and across a span of three months, and is dominated by relatively high-abundance T cell clones. Our results suggest the existence of a broad alloreactive T cell repertoire consisting mainly of expanded memory T cells which remain stable over time. The presence of a large, stable population alloreactive T cells should allow for long-term tracking of alloreactive clones in patients receiving allografts.
Granzyme B plays a differential role in CD4+ T cell subsets versus CD8+ T cells in mediating the graft-versus-tumor effect and graft-versus-host disease (TRAN3P.888)
Authors: Du Wei , Leigh Nicholas , Bian Guanglin , Qiu Jingxin , Chen George , Liu Hong , Hahn Theresa , McCarthy Philip. , Cao Xuefang 
Published: January 2023

Abstract
After allogeneic hematopoietic cell transplantation, donor-derived T cells can attack host tumor cells, producing a favorable graft-vs.-tumor (GVT) effect. However, graft-vs.-host disease (GVHD) may develop when donor T cells attack normal host tissue. We previously reported that while granzyme B (GzmB) is required for CD8+ T cells to cause GVHD, GzmB-mediated damage of CD8+ T cells impairs the GVT effect. In this study, we examined the role of GzmB in GVT and GVHD mediated by CD4+ T cell subsets, using an established A20 tumor model and bioluminescence imaging to monitor tumor growth in BALB/c hosts. We found that CD4+CD25+ regulatory T cells (Tregs) derived from C57BL/6 donors significantly suppressed GVT activity mediated by both CD8+ T cells and CD4+CD25- effector T cells (Teffs). However, GzmB-/- Tregs were as equally suppressive as WT Tregs, consistent with our previous report that GzmB is not required for Treg-mediated suppression of GVHD. Nonetheless, GzmB-/- Teffs were less effective in controlling tumor growth than WT Teffs; but surprisingly, GzmB-/- Teffs caused more severe GVHD than WT Teffs. Our data reveal that GzmB is essential for CD4+CD25- Teff-mediated optimal GVT effect but diminishes the ability of Teffs to cause GVHD, which contradicts its role in CD8+ T cells where GzmB is essential to cause GVHD but impairs GVT effect. These differential roles suggest that targeting GzmB in selected T cell subsets may provide a strategy to separate GVT from GVHD.
Ethyl pyruvate, an inhibitor of high-mobility group box 1 (HMGB1) release, modulates dendritic cell activation and survival (TRAN3P.897)
Authors: Chakhtoura Marita , Chain Robert , Varghese Linda , Gallucci Stefania 
Published: January 2023

Abstract
Dendritic cells (DCs) activated by stimulatory danger signals initiate the detrimental anti-graft immune responses that lead to transplant rejection. High-mobility group box1 (HMGB1) is a nuclear DNA-binding protein that signals danger if released, contributing to DC activation and acute allograft rejection. Preventing HMGB1 release is a way to block its effects, promoting less active DCs. Here we assessed the effects of ethyl pyruvate (EP), proposed to potentially prevent HMGB1 release from cells, on DC responses. We found that EP (10mM) abolished pro-inflammatory cytokine (IL-12, IL-6 and TNFa) and IL-10 induction by the strong DC stimulus LPS in myeloid bone marrow-derived dendritic cells (BMDCs). EP also significantly decreased BMDC surface MHC expression, suggesting a possible role in inhibiting antigen presentation. We also determined EP effects on BMDC viability and found that EP strongly inhibited in vitro DC activation without inducing death. Furthermore, EP could prevent LPS-induced apoptosis that normally occurs 72h post-stimulation. Moreover, EP suppressed DC responses to other TLR stimuli including CpGs and R848. Our results indicate that EP inhibits most of the biological responses of DCs to LPS and other TLR stimuli. We are investigating the link between EP and HMGB1 release prevention from BMDCs. Testing the effect of HMGB1 blockade on DC activation will highlight a novel target to induce tolerogenic DCs and prolong solid organ graft survival.
Pancreatic islets engineered to display on their surface an apoptotic form of SA-FasL induce localized tolerance via phagocytes/TGF-β/Treg axes (TRAN3P.866)
Authors: Woodward Kyle , Yolcu Esma , Zhao Hong , Bandura-Morgan Laura , Askenasy Nadir , Shirwan Haval 
Published: January 2023

Abstract
Islets engineered to display on their surface a FasL protein chimeric with streptavidin (SA-FasL) induce robust localized allotolerance under the transient cover of rapamycin. We hypothesized that immunomodulation with SA-FasL sets in action a regulatory circuit that involves apoptosis of alloreactive T cells within the graft microenvironment, clearance of apoptotic bodies by phagocytes, secretion of TGF-β, and generation/expansion of Tregs. This study was designed to test this hypothesis. SA-FasL-engineered islets resulted in alloreactive T cell depletion within the graft-draining lymph nodes with minimal impact on the cells in the spleen. Depletion of phagocytes one day prior to transplantation resulted in prompt rejection of SA-FasL-engineered islets (n=7). Similarly, treatment of graft recipients with a blocking Ab to TGF-β early (day 0, n=7), but not late (&gt; 100 days, n=5), post-Tx resulted in prompt rejection of long-term allogeneic SA-FasL-islet grafts. In marked contrast treatment with a blocking Ab to IL-10 had no effect on tolerance induction. F4/80+ cells in the graft were shown to co-localize with TGF-β 5 days post-Tx, suggesting that TGF-β in the graft produced by macrophages may induce tolerance. Taken together, these studies demonstrate that immunomodulation with SA-FasL-engineered pancreatic islets results in a localized immunoregulatory cascade driven by SA-FasL-mediated apoptosis of alloreactive T cells, phagocytes, and TGF-β that confers tolerance.
Friend or foe? How an invertebrate distinguishes self from non-self in a natural histocompatibility interaction. (TRAN3P.912)
Authors: Karadge Uma , Gosto Minja , Nicotra Matthew 
Published: January 2023

Abstract
The existence of sophisticated self/non-self systems is not limited to vertebrates. Several colonial marine invertebrates have evolved systems that distinguish between conspecifics via cell-cell contact. This phenomenon, known as allorecognition, occurs when colonies encounter each other as they grow across their substrate. Compatible colonies fuse or co-exist, while incompatible colonies reject and often aggressively compete for space. In all taxa studied to date, allorecognition phenotypes are determined by highly polymorphic loci, which ensure that colonies are only compatible with themselves or close kin. How allorecognition molecules achieve this specificity remains unknown. Allorecognition in Hydractinia is controlled by at least two genes, alr1 and alr2, which encode highly polymorphic transmembrane proteins similar to immunoglobulin superfamily molecules. Colonies with matching alleles at alr1 and alr2 fuse, while colonies with no matching alleles reject. Using in vitro assays with recombinant proteins, we demonstrate that alr1 alleles bind to themselves but not to other alr1 alleles. We suggest that, in vivo, compatibility between colonies is also determined by allele-specific homophilic binding of alr proteins. Since fusion is rare in nature and single populations contain hundreds of unique alr alleles, the Hydractinia allorecognition system appears to be based on a biophysical mechanism with unprecedented allelic diversity and specificity.
A bacterial protein supresses direct allorreactivity through induction of MARCH-I and downregulation of MHC class II in skin migratory dendritic cells (TRAN3P.913)
Authors: Bonorino Cristina , Borges Thiago 
Published: January 2023

Abstract
We have elucidated a major innate mechanism leading to the immunosuppressive actions of a member of the Hsp70 family, the DnaK of Mycobacterium tuberculosis. Treatment of skin allografts with Hsp70 lead to delayed acute rejection. This was mediated by skin CD103+ DCs, which migrated from the skin graft to the host DLN, resulting in decreased CD8+ and CD4+ T cell proliferation. Hsp70 modulated direct alloreactivity, signalling on donor DCs through TLR2 for the induction of IL-10, which resulted in MHC class II downregulation. Membrane-Associated Ring-CH I (MARCH-I), an ubiquitinator molecule responsible for downregulation of MHC II, was induced in DCs by Hsp70 via ERK, STAT3 and IL-10. This is a potent immunosuppressive effect that probably plays a role in immune evasion in vivo, and yet can be harnessed to manage graft rejection, without the burden of systemic immunosuppression for the host.
SA-FasL-engineered rat pancreatic islets induces robust localized tolerance through TGF-β/CD4+CD25+FoxP3+ Treg cells axis in mice (TRAN3P.867)
Authors: Zhao Hong , Yolcu Esma , Woodward Kyle , Ahamad Niloufar , Morgan Laura , Askenasy Nadir 
Published: January 2023

Abstract
Islets engineered to display on their surface an apoptotic form of FasL protein chimeric with streptavidin (SA-FasL) induce tolerance in the BALB/c-to-C57BL/6 allogeneic model. This study assessed the efficacy of this approach in a xenogeneic setting. WF rat islets were modified with biotin and engineered with SA-FasL protein. SA-FasL-islets survived indefinitely (n=10, &gt;200 days) in chemically diabetic C57BL/6 mice under transient cover of rapamycin. SA-engineered (n=5) or unmanipulated control (n=6) islet grafts were rejected acutely with MST=34.6±5.0 and 13.8±4.0, respectively. Tolerance was localized to the graft as long-term (&gt; 100 days) mice rejected a second set of unmodified graft transplanted under the contralateral kidney capsule without an impact on the survival of original graft. Flow-cytometry analysis and immunohistochemistry demonstrated significantly higher percentage of CD4+CD25+FoxP3+ Treg within the graft and graft-draining lymph nodes, but not spleen, of long-term recipients. Tolerant mice had decreased percentages of T cells expressing IL-2 and IFN-r as compared with rejecting animals. Treg cells depletion resulted in prompt graft rejection. Blocking of TGF-β in vivo during tolerance induction also resulted in the abrogation of graft survival. Testing the efficacy of this approach in a large animal model of xenotransplantation will be an important next step towards clinical translation of this novel tolerance concept.
FWGP activates the immune system (TUM7P.930)
Authors: Abuhay Mastewal , O'Donnell Robert , Ma Yunpeng , Xiong Chengyi , Tuscano Joseph 
Published: January 2023

Abstract
The immune system is an essential defense against malignancy; immunosuppression increases the risk of developing cancer. A better understanding of how to modulate the immune system may lead to new methods to prevent and treat disease. While recent developments in targeted immunomodulation have improved cancer treatments, most therapeutics are toxic, expensive, or have marginal efficacy. We developed a natural product derived from fermented wheat germ extract (FWGE) that acts as an immune modulator. Through further purification we identified protein components, (fermented wheat germ proteins, FWGP), that are more potent than FWGE. In vitro, FWGP was cytotoxic against B cancer cells with IC50s ranging from 10μg/ml to 17.5μg/ml. In vivo studies showed substantial efficacy without toxicity. We found that FWGP was a potent immune modulator. It increased CD4+ T cells, NK-T cells, & activated monocytes by 4, 10, & 50 folds, respectively, while decreasing CD8+ T cells. CD56-bright NK cells treated with FWGP increased compared to the heat-inactivated FWGP treatment, and NK-mediated IFN-γ production rose as [FWGP] increased. Also, FWGP (0.1 mg/ml) activated complement with 2-fold potency compared to either control or FWGE (55mg/ml). It enhanced ADCC as well; PBMCs treated with FWGP showed a dose dependent increase in killing of Ramos. FWGP’s in vivo efficacy could be due to its immune modulating effects and may be the basis for a non-toxic treatment for lymphoma and other malignancies.