bluuebunny/crossref_metadata_2025_split huggingface dataset:
Shared VH1-46 antibody gene usage in pemphigus vulgaris predicts antibody cross-reactivity to desmoglein 3 and rotavirus VP6 (HUM1P.271)
Authors: Cho Michael Jeffrey , Ellebrecht Christoph , Hammers Christoph , Sapparapu Gopal , Crowe James , Payne Aimee 
Published: January 2023

Abstract
Pemphigus vulgaris (PV) is a potentially fatal skin blistering disease caused by autoantibodies (autoAbs) to desmoglein 3 (Dsg3). Previously, we identified shared VH1-46 gene usage in the anti-Dsg3 repertoire among 4 PV patients via high throughput B cell receptor (BCR) cloning. Somatic mutation reversion studies indicated that VH1-46 BCRs require few to no acidic amino acid mutations to bind Dsg3, which may favor their early selection in the immune response. Interestingly, shared VH1-46 gene usage also occurs in the B cell response to rotavirus protein VP6, and similarly, few somatic mutations are required to bind VP6. We thus investigated whether Dsg3-reactive VH1-46 BCRs cross-react to VP6 which may explain the tolerance of these autoreactive clones. We screened ~108 combinatorial IgM clones from a PV patient, which identified 13 heavy chain CDR3 clonotypic families that cross-react to Dsg3 and VP6. 7/13 used VH1-46, indicating enrichment by Dsg3/VP6 antigen selection. ELISA and human skin immunofluorescence using purified soluble Abs confirmed VP6 and Dsg3 reactivity. Additionally, 3 of 5 VH1-46 anti-Dsg3 IgG clonotypes previously isolated from 3 different PV patients cross-reacted to VP6 in either their somatically mutated or unmutated form. Our data suggest that a VH1-46 B cell response to rotavirus may be an initiating event that develops into an autoimmune response to Dsg3 in susceptible individuals.
Development of novel sterol-based Liver X Receptor agonists as therapeutics for inflammatory diseases (HUM1P.307)
Authors: Yu Shan , Li Shangjin , Henke Adam , Welzel Gustav , Li Sijia , Cheng Bo , Evan Muse , Glass Christopher , Chatterjee Arnab , Roland Jason , Schultz Peter , Tremblay Matthew 
Published: January 2023

Abstract
Liver X Receptor (LXR), a nuclear hormone receptor, is an essential regulator of immune responses. Activation of LXR by synthetic agonists, including T0901317 and GW3965, attenuates progression of inflammatory diseases of animal models. However, the adverse effects of traditional LXR agonists in elevating liver lipids have impeded exploitation of this intriguing mechanism for chronic therapy. The endogenous LXR agonist desmosterol and its sterol-based analog DMHCA activate LXR without the lipogenic action in the liver - this inspired us to develop anti-inflammatory therapies based on these analogs. Here, we show DMHCA oral treatment in mice significantly reduce DSS colitis-induced body weight loss, which is accompanied by reduced expression of inflammatory markers in the large intestine. This reduced inflammation is associated with induced LXR target gene expression, suggesting DMHCA attenuating colitis is mediated, at least partially, through LXR activation. In addition, DMHCA treatment dramatically suppresses inflammatory cytokine expression in traumatic brain injury model. Importantly, in both disease models, DMHCA does not increase liver lipid or serum ALT levels, while T0901317 results in significant liver lipid accumulation and injury. The anti-inflammatory property of DMHCA is recapitulated in vitro in CD3/CD28 activated human PBMCs. Overall, these results provide evidence for the development of sterol-based LXR agonists as novel therapeutics for inflammatory diseases.
Inflammation and gut microbiota-induced hepatic lipogenesis drives metabolic syndrome in TLR5 deficient mice (HUM1P.310)
Authors: Singh Vishal , Chassaing Benoit , Zhang Limin , Yeoh Beng San , Xiao Xia , Harvatine Kevin , Ntambi James , Patterson Andrew , Gewirtz Andrew , Vijay-Kumar Matam 
Published: January 2023

Abstract
The gut microbiota plays a key role in host metabolism. Toll-Like Receptor 5 (TLR5), a flagellin receptor, is required for gut microbiota homeostasis. Accordingly, TLR5 deficient (T5KO) mice are prone to develop microbiota-dependent spontaneous colitis and metabolic syndrome. Herein, we investigated whether metabolic syndrome in T5KO mice correlates with hepatic dyslipidemia and inflammation. T5KO mice displayed elevated neutral lipids with substantial enrichment of C18:1 (n9) relative to wild-type littermates. Oleate enrichment of hepatic lipids was microbiota-dependent. Analyzing cecal contents via 1H NMR-based metabolomics revealed that T5KO mice exhibited elevated short chain fatty acids (SCFA) with a concomitant increase in colonic SCFA receptors and hepatic pro-inflammatory genes and lipogenic enzymes including stearoyl-CoA desaturase1 (SCD1). SCFA treatment further aggravated metabolic syndrome in T5KO mice. Interestingly, deletion of hepatic SCD1 not only prevented hepatic neutral lipid oleate enrichment but also ameliorated hepatic inflammation and metabolic syndrome in T5KO mice. Collectively, these results underscore the key role of the gut microbiota-liver axis in the pathogenesis of metabolic diseases.
Immune-related gene polymorphisms as risk factors for the development of Factor VIII inhibitors in hemophilia A patients: a sibling-pair association analysis (HUM1P.257)
Authors: Ferrante Andrea , Mosher Bryan , Hoffman Megan 
Published: January 2023

Abstract
Inhibitors are one of the most feared complications in hemophilia, occurring in 15-30% of individuals with severe forms of Hemophilia A Human Leukocyte Antigen (HLA) haplotype, and polymorphism of genes regulating the immune response have been indicated as risk factors for inhibitor development. However, it is still unclear the extent to which each of these genetic variants contribute to inhibitor formation as a function of the F8 gene mutation. Here, 96 patients clustered in 45 sibling-pairs and 2 sibling-triplets of which 15 are discordant and the remaining are concordant for inhibitors were considered in a case-control analysis. Next Generation Sequencing on patients’ genomic DNA was performed for detection of FVIII mutation, genotyping of HLA-DRB1 allele and for TNF-α, IL-10, CTLA4, DOCK2 and MAPK9. Our results demonstrate a higher frequency of DRB1*15 allele in inhibitor patients [odds ratio (OR) 2.57; P = 0.022]. In TNF-α, the A allele of the -308G&gt;A polymorphism was found with higher frequency in the inhibitor cohort. This finding was ore evident for the homozygous A/A genotype (7.7% vs 1.8% in inhibitor patient group vs. non-inhibitor respectively, p=0.028). For IL-10, the -1082G allele was observed more frequently in patients with inhibitors (59% vs. 37.5%; P = 0.0037). Our data validate the hypothesis that specific genomic haplotypes affect the development of FVIII inhibitors and can support inhibitor prediction for prognostic and therapeutic applications.
Chronic upper airway inflammation is characterized by an extrafollicular B cell response and elevated antibody production in humans (HUM1P.260)
Authors: Hulse Kathryn , Tan Bruce , Kasjanski Richard , Van Roey Griet , Zhang Chaoying , Norton James , Carter Roderick , Suh Lydia , Kato Atsushi , Kern Robert , Conley David , Peters Anju , Grammer Leslie , Harris Kathleen , Schleimer Robert 
Published: January 2023

Abstract
Chronic rhinosinusitis (CRS) is an upper airway disease that affects 30 million Americans and a subgroup of CRS patients also has nasal polyps (CRSwNP). In CRSwNP patients, inflamed sinus tissues can be removed as part of treatment, which provides a unique model system to investigate mechanisms of human mucosal inflammation. We have previously reported local elevations in NP of B cells, antibodies (Ab), and expression of Epstein-Barr virus-Induced protein 2 (EBI2), a marker of extrafollicular plasma cells (PC). To characterize the mechanisms mediating these responses, we assessed B cells from fresh tissue (d0) or after 4 days of culture (d4) from NP, uncinate tissue (UT) or tonsils from CRS and control subjects by flow cytometry. We measured Ab production by multiplex array and assessed formation of B cell follicles by immunohistochemistry. We assessed the Ab repertoire by DNA deep sequencing and by analysis of Ab genes from single cells. We found that NP uniquely supported the accumulation of PC (p&lt;0.01) and Ab production (p&lt;0.05) in vitro. Plasmablasts (PB) and PC were Ki67+, and EBI2 expression was highest on PB. We found no evidence for increased B cell follicles in NP compared to control UT (n&gt;10/group). Ab from NP had fewer CDR3 mutations than Ab from UT (p&lt;0.05), but more mutations than Ab from peripheral blood (p&lt;0.05). These data suggest that there is a strong extrafollicular B cell response in NP that may drive Ab production during chronic airway inflammation.
The TNF-family ligand TL1A and its receptor DR3 promote T cell mediated allergic immunopathology by enhancing differentiation and pathogenicity of IL-9 producing T cells (HYP2P.333)
Authors: Meylan Francoise , Richard Arianne , Tan Cuiyan , Hawley Eric , Gomez-Rodriguez Julio , Goswami Ritobrata , Yang Xiangping , Cruz Anthony , Penumetcha Pallavi , Hayes Erika , Pelletier Martin , Gabay Odile , Walsh Matthew , Ferdinand John , Keane-Myers Andrea , Choi Yongwon , O'Shea John , Al-Shamkhani Aymen , Kaplan Mark , Gery Igal , Siegel Richard 
Published: January 2023

Abstract
The TNF-family cytokine TL1A (Tnfsf15) costimulates T cells and type 2 innate lymphocytes (ILC2) through its receptor DR3 (Tnfrsf25). DR3-deficient mice have reduced T cell accumulation at the site of inflammation, and reduced ILC2-dependent immune responses in a number of models of autoimmune and allergic diseases. In allergic lung disease models, immunopathology and local Th2 and ILC2 accumulation is reduced in DR3 deficient mice despite normal systemic priming of Th2 responses and generation of T cells secreting IL-13 and IL-4, prompting the question of whether TL1A promotes the development of other T cell subsets that secrete cytokines to drive allergic disease. Here we find that TL1A potently promotes generation of murine T cells producing IL-9 (Th9) by signaling through DR3 in a cell-intrinsic manner. TL1A enhances Th9 differentiation through an IL-2 and STAT5-dependent mechanism, unlike the TNF-family member OX40, which promotes Th9 through IL-4 and STAT6. Th9 differentiated in the presence of TL1A are more pathogenic, and endogenous TL1A signaling through DR3 on T cells is required for maximal pathology and IL-9 production in allergic lung inflammation. Taken together, these data identify TL1A-DR3 interactions as a novel pathway that promotes Th9 differentiation and pathogenicity. TL1A may be a potential therapeutic target in diseases dependent on IL-9.
Pentraxins interact with misfolded proteins and inhibit aberrant innate immune activation (INC1P.361)
Authors: Dorta-Estremera Stephanie , Cao Wei 
Published: January 2023

Abstract
Amyloid deposition is linked to different diseases, including Alzheimer’s disease and systemic amyloidosis. How misfolded proteins, including amyloid and its in-process products, are controlled by the host remains obscure. Pentraxins are multimeric pattern recognition proteins that bind to diverse ligands and promote the clearance of microbes. Serum amyloid P-component (SAP), a short chain pentraxin, associates with amyloid deposits in patients. We hypothesize that pentraxins may interact with different forms of misfolded proteins and modulate their biological impact. We found that human SAP readily bound to all types of amyloids, including protein-only, DNA-containing or heparin-containing fibrils. This interaction depended on the presence of calcium. Interestingly, both SAP and C-reactive protein (CRP), another pentraxin, bound to soluble amyloid precursor (AP) protein in the absence of divalent cations. Furthermore, SAP decamer recognized AP whereas aggregated SAP preferentially associated with amyloid. The misfolded structure in AP was necessary for binding to SAP and CRP. DNA-containing amyloid (A-DNA) triggers type I interferon (IFN-I) production by plasmacytoid dendritic cells (pDCs) and promotes lupus-like disease. Binding of SAP to A-DNA inhibited the production of IFN-I by human pDCs. Altogether, our data suggest that pentraxins differentially interact with various forms of misfolded proteins and modulate their ability to stimulate aberrant innate immune activation.
The RiboPuromycylation Method Reveals Rapid IL12-Dependent Antigen-Independent Activation of Bone Marrow Virtual Memory T cells (INC1P.403)
Authors: Seedhom Mina , David Alexandre , Hickman Heather , Bennink Jack , Yewdell Jonathan 
Published: January 2023

Abstract
Protein translation is an essential and intricately regulated cellular process that can potentially be used to monitor immune cell activation. We extended the RiboPuromycylation Method (RPM) to measure protein translation in vivo. Unexpectedly, 24 h following either vaccinia virus (VACV) or vesicular stomatitis virus (VSV) infection, 15-30% of bone marrow (BM) CD4+ and CD8+ T cells, all CD44+, increase protein translation. This is days before protein translation is increased in T cells in the draining lymph node or spleen. Bystander BM but not splenic OT-1 T cells demonstrate increased protein translation after VACV infection. Early virus-induced BM T cell translation is absent in IL-12 -/- mice. In BM cultures from naïve mice, addition of IL-12, and especially IL-12 with IL-18 increase T cell translation within hours, with IL1-2 and IL-18 eventually leading to T cell division. Our findings indicate that the BM is a unique environment where bystander T cells are rapidly activated by innate immune response to viruses. We hypothesize that this may allow memory and memory-like T cells to be activated in the absence of cognate antigen to facilitate their functionality when they encounter the cognate pathogen.
Regulation of the gut microbiota and intestinal homeostasis by phagocyte NADPH oxdiase (INC1P.351)
Authors: Zeng Melody 
Published: January 2023

Abstract
The phagocyte NADPH oxidase generates reactive oxygen species (ROS) for microbial killing, and its impairment gives rise to Chronic Granulomatous Disease (CGD), a primary immunodeficiency characterized by recurrent infections and granulomatous lesions. Mutations in Nox2 (Cybb), a critical subunit of the oxidase, contribute 66% of CGD. In our study, we found spontaneous inflammation in the intestine of naïve Cybb-/- mice, indicated by higher levels of IL-1b, IL-6 and TNFα in lamina propria macrophages, as well as increased levels of IFNγ and IL-17A in CD4+ T cells. In addition, Klebsiella pneumoniae, a bacterium frequently found in granulomatous lesions in CGD patients, were detected in the spleens and mesenteric lymph nodes (MLNs) of naïve Cybb-/- mice, and IP administration of this pathobiont strain of K. pneumonia led to rapid mortality of wild-type mice. Oral administration of this K. pneumoniae induced intestinal inflammation in wild-type and Cybb-/- mice, but translocation of K. pneumoniae to MLNs was drastically enhanced in Cybb-/- mice and bone marrow chimeras with a neutrophil-specific deficiency in NOX2. Thus, our results demonstrate the importance of neutrophil-derived ROS in restricting dissemination of pathobionts from the intestine under steady-state conditions. Therefore, our study underscores a critical role for the NADPH oxidase in regulating intestinal homeostasis and provides insights into intestinal complications in CGD patients.
IL-6 regulates pro- and anti-inflammatory cytokine production and mortality of Mycobacterium tuberculosis infected type 2 diabetic mice (INC2P.418)
Authors: Cheekatla Satyanarayana , Tripathi Deepak , Venkatasubramanian Sambasivan , Paidipally Padmaja , Welch Elwyn , Tvinnereim Amy , Kornfeld Hardy , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
In this study, to determine whether diabetic (DM) mice are susceptible to Mycobacterium tuberculosis (Mtb) infection, we developed experimentally induced diabetes in wild type C57BL/6 mice with streptozotocin & nicotinamide. We then infected non-DM and DM mice with Mtb H37Rv by aerosol. Around 50% of Mtb infected DM (p=0.05) and 25% of uninfected DM (p=0.07) mice died in 5 to 6 months compared to no deaths in infected control mice. Six months after Mtb infection, there was a statistically significant but marginal increase in bacterial burden in the lungs of Mtb infected DM mice compared to non-DM infected mice (3.1 ± 0.4 x106 vs 0.9 ± 0.1 x 106, p=0.001) suggesting increased mortality was not due to increased bacterial burden. Real time PCR analysis of Mtb infected DM lungs indicated significantly increased pro- and anti-inflammatory cytokine gene expression compared to uninfected DM and infected control mice, including increased levels of IL-6 mRNA. Anti-IL-6 antibody treatment of Mtb infected DM mice enhanced the survival (100% vs 60% survival in isotype control antibody treated, p=0.02) and reduced pro- and anti-inflammatory cytokine production. Dendritic cells (DC), but not other immune cells were the major source for IL-6 in Mtb infected DM mice. NK cells of Mtb infected DM mice further enhanced IL-6 production by autologous DCs. Our results suggest NK-DC interaction enhance IL-6 production which drives the immune pathology and mortality of Mtb infected diabetic mice
Novel anti-Mer antibodies in macrophage phenotype analysis (INC2P.419)
Authors: Stepniak Dariusz , Atteberry Brandi , Li Shourong , Baterina Oscar , Oxford Natalie , Miranda Cherlyn , Miletic Ana , Schrantz Nicolas , Huang Qiulong , Funatake Castle , Just Margaret 
Published: January 2023

Abstract
Mer is a member of the TAM family of tyrosine kinase receptors that also includes Axl and Tyro3. Mer is expressed on a subset of anti-inflammatory macrophages and is involved in the removal of apoptotic cells. The engulfment of apoptotic cells relies on two soluble accessory molecules Protein S and Gas6 that bind to phosphatidylserine, which is found on the outer leaflet of the plasma membrane of cells undergoing apoptosis. Upon binding these ligands, Mer undergoes autophosphorylation at multiple tyrosine residues that activate the PI3K and Akt pathways. This then leads to the phagocytosis and clearance of apoptotic cells and also results in the direct inhibition of TLR-induced production of pro-inflammatory cytokines. Deficiency of Mer causes general autoimmunity, inflammation, and accumulation of apoptotic bodies. Because Mer expression strongly correlates with macrophage function, It can be used for phenotyping macrophage subpopulations. Here we use novel anti-Mer antibodies in flow cytometric analysis of macrophages and corroborate the expression of this receptor and its importance in distinguishing functional macrophage subsets.
Essential role for CCR2+ inflammatory monocytes in early and late control of persistent Salmonella infection (INC2P.412)
Authors: Van der Velden Adrianus , Tam Jason , Bliska James 
Published: January 2023

Abstract
Monocytes play a critical role in innate and adaptive immunity. Monocytes originate from progenitors in the bone marrow and traffic via the bloodstream to peripheral tissues, where they can differentiate into protective cell types such as macrophages and dendritic cells. Emigration of Ly6Chi monocytes from the bone marrow is dependent on CC-chemokine receptor 2 (CCR2) and largely mediated by CC-chemokine ligand 2 (CCL2, also referred to as MCP1). We previously showed that large numbers of Ly6Chi Ly6G- CD11b+ monocytes accumulate and persist in tissues of mice infected with the bacterial pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium). Here, we report that Ccr2 is required and Ccl2 is important for the recruitment of Ly6Chi Ly6G- CD11b+ monocytes into tissues during persistent S. Typhimurium infection of hemizygous C57BL/6J Nramp1G169 mice, which are phenotypically Nramp1+. We show that Ccr2- and Ccl2-deficient mice are more susceptible to S. Typhimurium infection than wild-type mice, with Ccr2-deficient mice being more susceptible to S. Typhimurium infection than Ccl2-deficient mice. We also show that depletion of CCR2+ cells during the first or third week of S. Typhimurium infection increases susceptibility to salmonellosis. These results indicate that CCR2+ cells, including Ly6Chi Ly6G- CD11b+ monocytes, play an essential role in early and late control of S. Typhimurium infection.
TRAF6 and TAK1 control signals for a senescence response by an endosomal NK cell receptor (INC2P.414)
Authors: Rajagopalan Sumati , Lee Elizabeth , DuPrie Matthew , Long Eric 
Published: January 2023

Abstract
The endosomal innate receptor CD158d (KIR2DL4) induces cellular senescence in human NK cells in response to soluble ligand (HLA-G or agonist antibody). These senescent NK cells display a senescence-associated secretory phenotype (SASP) and their secretome promotes vascular remodeling and angiogenesis. To understand how CD158d initiates signaling for a senescence response, we mapped the region in its cytoplasmic tail that controls signaling. We identified a conserved TRAF6 binding motif, which was required for CD158d-induced NF-κB activation and IL-8 secretion, for TRAF6 association with CD158d, and for TRAF6 recruitment to CD158d+ endosomes in transfected cells. The adaptor TRAF6 is known to couple proximal signals from receptors such as endosomal TLRs and CD40 through the kinase TAK1 for NF-κB-dependent pro-inflammatory responses. siRNA-mediated silencing of TRAF6 and TAK1, and inhibition of TAK1 blocked CD158d-dependent IL-8 secretion. Stimulation of primary, resting NK cells with soluble Ab to CD158d, induced TRAF6 association with CD158d, induced TAK1 phosphorylation, and inhibition of TAK1 blocked the CD158d-dependent reprogramming of NK cells that produces the SASP signature. Our results reveal that a prototypic TLR and TNF-receptor signaling pathway is used by a killer cell immunoglobulin-like receptor that promotes secretion of pro-inflammatory and pro-angiogenic mediators as part of a unique senescence phenotype in NK cells.
Mast cell-derived histamine is necessary for platelet-activating factor mediated systemic immunosuppression (IRC4P.606)
Authors: Ocana Jesus , Yi Quiaofang , Yao Yongxue , Bryce Paul , Kaplan Mark , Travers Jeffrey 
Published: January 2023

Abstract
Platelet-activating Factor (PAF) activates many cell types via the G-protein coupled PAF-receptor (PAF-R). Systemic PAF-R activation exerts acute pro-inflammatory, as well as delayed systemic immunosuppressive effects in vivo. PAF enzymatic synthesis and degradation is tightly regulated, but reactive oxygen species from oxidative stressors, such as UVB, chemotherapy and cigarette smoke, generate PAF-like species via the unregulated oxidation of membrane lipids. The systemic immunosuppressive effects of these pro-oxidative stressors are due to PAF-R signaling. This work establishes the role of the mast cell (MC) and the soluble mediators involved in PAF-R-mediated immunosuppression. Using a contact hypersensitivity model, we show that PAF loses its immunosuppressive effects in MC-deficient (Wsh) mice, a phenotype rescued by dermal reconstitution of Wsh mice with WT bone marrow derived MCs. Use of PAF-R-deficient Wsh mice suggests that the MC PAF-R is necessary and sufficient to mediate PAF-R systemic suppression, in a process attenuated by COX-2 and TGF-β inhibitors. Finally, by transplanting histamine (histidine decarboxylase-null) deficient MCs, we demonstrate that this pathway requires MC-derived histamine release. These studies fit with the hypothesis that MC PAF-R activation with resultant histamine release mediates PAF-R systemic immunosuppression.