bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autoantibody-positive healthy individuals display unique immune profiles that regulate autoimmunity (HUM3P.244)
Authors: Slight-Webb Samantha , Lu Rufei , Ritterhouse Lauren , Maecker Holden , Fathman C. , Merrill Joan , Guthridge Joel , James Judith 
Published: January 2023

Abstract
Antinuclear antibodies (ANA) are detected in up to 25% of the population, yet only 5-8% develop autoimmunity. Immunological differences between ANA+ healthy individuals and SLE patients may give us insight into mechanisms by which ANA+ individuals avoid autoimmunity or transition to SLE. European-American ANA+ individuals (n=31) were matched to ANA- controls and SLE patients. Serum cytokine profiles, lymphocyte subset frequency, and lymphocyte reactivity were analyzed. Numerous pro-inflammatory cytokines including IFNγ, IL-2, IL-5 and TNFα were elevated in the serum of ANA+ healthy individuals compared to ANA- controls; however, SLE patients had even higher cytokine levels compared to ANA+ healthy individuals (p&gt;0.05). Interestingly, Type I IFNs, BLyS, IL-12p40 and SCF were only increased in SLE patients (p&lt;0.05). Further, IL-1RA was decreased in both ANA+ and SLE individuals (p&lt;0.0001). Total monocytes and memory B cells, were elevated in ANA+ individuals compared with ANA- controls while regulatory CD4+ T cells were decreased (p&lt;0.05). Further, monocytes and B cells from ANA+ individuals had increased levels of pSTAT1 and pSTAT3 following IFN-α stimulation compared to controls (p&lt;0.05). These data reveal ANA+ healthy individuals exhibit similar features to SLE patients with increased inflammation and loss of immune tolerance, but differ by the absence of elevated Type I IFNs, BLyS, IL-12p40 and SCF and severely decreased IL-1RA that contribute to disease pathogenesis.
CD3/SLAMF3-mediated co-stimulation promotes activation of the IL-2/IL-2R/STAT5 pathway and iTreg differentiation in naïve CD4+ T cells - implications for systemic lupus erythematosus (HUM3P.246)
Authors: Comte Denis , Kis-Toth Katalin , Karampetsou Maria , Yoshida Nobuya , Solomon Julie , Tsokos George 
Published: January 2023

Abstract
We examined SLAMF3-mediated co-stimulation on IL-2 response of CD4+ T cells from SLE patients. Stimulation of naïve CD4+ T cells with aCD3/aSLAMF3 upregulated surface CD25 (IL-2Ra) and CD122 (IL-2Rb), at higher level compared to aCD3/aCD28 activation. aCD3/aSLAMF3 co-stimulation of control naïve CD4+ T cells led to increased pSTAT5 levels compared to aCD3/aCD28 activation Although naïve CD4+ T cells from SLE patients displayed decreased pSTAT5 levels compared to control donors upon aCD3 or aCD3/aCD28 stimulation, co-stimulation of SLE T cells with aSLAMF3 restored pSTAT5 to normal levels. Total STAT5 levels remained unaltered after T cell activation. Exogenous IL-2 increased proliferation of aCD3/aSLAMF3-activated CD4+ T cells compared to aCD3/aCD28 stimulation. Naïve CD4+ T cells activated with aCD3/aSLAMF3 under iTreg polarizing condition expressed significantly higher levels of FoxP3, CD25 and CTLA-4 compared to aCD3/aCD28 activation. In conclusion, SLAMF3-mediated co-stimulation enhances CD4+ T cell response to IL-2 in SLE patients by acting on the IL-2/IL-2R/STAT5 signaling pathway, increases naïve CD4+ T cell proliferation in response to exogenous IL-2 and can promote iTreg phenotype generation. Of note, SLAMF3-mediated co-stimulation does not increase IL-2 by CD4+ T cells per se. Thus, aSLAMF3-based Ab therapy may emerge as a promising therapeutic agent in SLE by enhancing response to IL-2 and promoting iTreg differentiation.
ROCK2 signaling is required for development and function of T follicular helper cells (HUM3P.247)
Authors: Weiss Jonathan , Chen Wei , Nyuydzefe melanie , Flynn Ryan , Tonra James , Marusic Suzana , Blazar Bruce , Waksal Samuel , Zanin-Zhorov Alexandra 
Published: January 2023

Abstract
PD1+CXCR5+ T follicular helper (Tfh) cells are essential for generating high-affinity antibodies and B cell memory. But aberrant Tfh activity can drive autoimmune disease such as systemic lupus erythematosus (SLE) which is characterized by high levels of auto-antibodies and increased frequencies of IL-21+ T cells. Rho-associated kinase (ROCK) 2 was recently shown to be critical in regulating IL-21 secretion in both mice and humans. Here, we show that the PD1+CXCR5+ subset is a major source of IL-21 in human CD4+ T cells stimulated in vitro. By using KD025, a Kadmon orally available, selective ROCK2 inhibitor, we attenuated the development of PD1+CXCR5+ T cells by over 60% and the perpetuation of existing Tfh cells in a dose dependent manner. Targeting of ROCK2 concurrently regulates STAT3/STAT5 phosphorylation and transcription activity followed by a reduction of Bcl6 and up-regulation of Blimp1 protein levels in human T cells. Oral administration of KD025 reduced the percent of both Tfh and plasma B cells in spleens by more than 2 fold as well as germinal center size in the Mrl/lpr murine model of SLE. KD025 treated animals had a substantial improvement in both histological and clinical scores complimented by a reduction in pSTAT3 and BCL6 expression in addition to an increase in STAT5 phosphorylation. This data suggests that ROCK2 controls the function of Tfh cells and underscores the therapeutic potential of targeted ROCK2 inhibition in the clinic.
The impact of obesity on human γδ T cell function and homeostasis (HUM4P.271)
Authors: Dutt Shelley , Costanzo Anne , Han Peggy , Taylor Kristen , Fujioka Ken , Jameson Julie 
Published: January 2023

Abstract
CDC estimate the rate of obesity is on the rise, currently impacting one-third of the U.S. population. Obese patients exhibit side effects that include type 2 diabetes, chronic wounds and an increased vulnerability to infection. It is critical to understand how obesity impacts immune responses that affect these processes and create effective vaccines or antiviral treatments for obesity. We examined PBMC from donors with a lean BMI &lt;25 or obese BMI &gt;30. T cells were studied for responses to influenza virus A/PR/9/38 infection by monitoring upregulation activation markers and cytokine production, using flow cytometry. From these studies we have determined that obese donors have quantifiably fewer γδ T cells, which has implications for rapid immune responses to infection. γδ T cells are normally activated upon contact with infected influenza virus cells, however our findings show that IFN-γ production by γδ T cells is reduced in obese patients. In contrast there was no significant difference in the expression of cytotoxic markers such as granzyme B and NKG2D between lean and obese patients. Due to our finding, IL-2Ra is reduced on the cell surface and IL-2 is able to restore γδ T cell antiviral function,we hypothesize that γδ T cells in obese patients are lacking key T cell specific growth factor signals. Our findings show that gd homeostasis and antiviral function becomes impaired with obesity contributing to the increased susceptibility to infection found in obese subjects.
STAT5 tetramer formation is critical for mast cell function (HYP4P.312)
Authors: Kiwanuka Kasalina , Lin Jian-Xin , Leonard Warren , Ryan John 
Published: January 2023

Abstract
Signal transducers and activators of transcription (STATs) are latent transcription factors that are vital for cell survival, proliferation and function. Activated STAT proteins dimerize and translocate to the nucleus, where they bind to high-affinity motifs to modulate gene expression. These dimers can also form tetramers through an N-terminal domain-mediated oligomerization process, and bind to low-affinity tandem motifs. We previously generated Stat5a-Stat5b double knock-in (DKI) mice in which STAT5 proteins are phosphorylated and can form dimers but not tetramers. We have now found that bone marrow-derived mast cells from Stat5 DKI mice express FceRI and c-Kit at levels comparable to those from wild-type mice. However, these mast cells are defective in IgE-induced cytokine and chemokine production, and exhibit less SCF-mediated migration in vitro. Similarly, IgE-mediated passive systemic anaphylaxis is also altered in Stat5 DKI mice. Unraveling the role of STAT5 tetramers may reveal novel insights into mast cell function in allergic and inflammatory diseases.
The role of 1a,25-dihydroxyvitamin D3 in IgE-mediated mast cell activation (HYP4P.313)
Authors: Baker Bianca , Clark Nicholas , Boyan Barbara , Schwartz Zvi , Ryan John 
Published: January 2023

Abstract
As the prevalence of allergic diseases such as allergic asthma and rhinitis continues to increase, there is growing interest in novel approaches to blunt the inflammatory processes. IgE-mediated mast cell activation results in the release of histamine and other inflammatory mediators. Vitamin D has been linked to allergic disease, with vitamin D deficiency being linked to increased risk for development and severity of allergic disease. We assessed the effects of the Vitamin D metabolite 1a,25-dihydroxyvitamin D3 (1,25D3) on IgE-mediated mast cell activation in vitro. We find that 1,25D3 suppresses IgE-mediated inflammatory cytokine and chemokine production. Surprisingly, these effects do not require expression of the canonical receptor for 1,25D3 (VDR), as wild type and VDR KO mast cells are similarly suppressed by 1,25D3. Additionally, these effects appear to be somewhat stimulus-specific, as IL-33-mediated mast cell activation s not inhibited by 1,25D3. We postulate that the chaperone protein Pdia3, an alternative plasma membrane-associated receptor for 1,25D3, may be responsible for the VDR-independent, anti-inflammatory effects on IgE-mediated mast cell activation. Our data depict novel evidence that 1,25D3 can attenuate the pro-inflammatory mast cell response, and emphasize the possible utility of vitamin D supplementation for treatment of allergic and inflammatory diseases.
Augmentation of local granulopoiesis via TLR2 signaling to promote resolution of S. aureus-infected wounds (INC4P.322)
Authors: Falahee Patrick , Anderson Leif , Dahmubed Delsheen , Simon Scott 
Published: January 2023

Abstract
Staphylococcus aureus is the most frequent cause of skin and soft tissue infections seen in emergency departments across the United States and the increasing global prevalence of virulent antibiotic resistant strains such as Methicillin-resistant S. aureus (MRSA) represents a major public health concern. Polymorphonuclear leukocytes (PMN), or neutrophils, are first-line effector cells of the innate immune system that play a central role in S. aureus clearance. Defects in PMN number and function secondary to comorbidities such as diabetes or myeloablative chemotherapy frequently contribute to poor outcomes in patient populations. Using a transgenic mouse model of S. aureus-infected skin wounds, we recently reported on a novel innate immune mechanism whereby a subset of hematopoietic stem and progenitor cells (HSPC) traffic to the site of infection, undergo local granulopoiesis via a TLR2 and MyD88 signaling pathway, and play a critical role in the resolution of infection. Local produced PMN exhibit enhanced antibacterial function compared with those produced in bone marrow. We examined whether this pathway for extramedullary granulopoiesis is conserved in humans and found that treatment of bone marrow-derived CD34+ cells with the TLR2 agonist Pam3CSK4 drives efficient expansion of functional PMN in vitro. Studies are underway to improve the expansion efficiency of HSPC into PMN in order to study their impact on persistent and chronic MRSA infection in mouse wounds.
Manipulation of macrophage autophagy by Coxiella burnetii (INC4P.336)
Authors: Winchell Caylin , Graham Joseph , Voth Daniel 
Published: January 2023

Abstract
Q fever is an emerging zoonosis caused by the intracellular bacterium, Coxiella burnetii. Disease can present as an acute illness with debilitating flu-like symptoms or a chronic illness resulting in life-threatening endocarditis. Alveolar macrophages are the primary target cell for infection in which C. burnetii traffics through the phagolysosomal pathway. Upon fusion with lysosomes, C. burnetii uses a type IV secretion system (T4SS) to modify and expand this compartment into a replication niche termed the parasitophorous vacuole (PV). Autophagy is a eukaryotic process in which portions of bulk cytoplasm or specifically selected cargo are enclosed by autophagosomes and trafficked to lysosomes for degradation. Macrophages also use autophagy for clearance of bacteria and inflammasome regulation. We previously showed that recruitment of autophagosomes to the PV is dependent on T4SS activity. The cargo receptor p62 is also recruited to PV, therefore we investigated the role of p62 during infection. We found that p62 interacts with distinct host proteins during infection, including the cargo receptor NBR1. Additionally, we discovered a potential C. burnetii effector that may modulate interactions with autophagosomes. Finally, to assess the role of p62 in the innate immune response, we initiated studies to determine interactions with inflammasome components. These studies are defining a novel role for eukaryotic cargo receptors in intracellular bacterial pathogenesis.
A protective role for splenic neutrophils against Streptococcus pneumoniae infection (INC4P.338)
Authors: Deniset Justin , Kubes Paul 
Published: January 2023

Abstract
The spleen is integral for protection against infections to encapsulated bacteria, such as S.pneumoniae. A recently identified splenic resident neutrophil subset has been proposed to contribute to this protection by modulating the antibody production of marginal zone B cells. To date, little is known regarding the function and behavior of these neutrophils in vivo and whether they actively contribute to protection against infection. Employing intravital microscopy we aimed to characterize splenic neutrophil behavior and function. Under basal conditions, two subsets of neutrophils are observed, a circulating phenotype and a resident phenotype. Infection with S. pneumonia stimulates a robust recruitment of neutrophils to the spleen and an enhanced localization within the marginal zone. This recruitment is accompanied by neutrophil activation typified by a reduction in cellular movement. Neutrophil depletion within the spleen during infection resulted in a reduction in bacterial clearance and a significant decrease in survival. Depletion of marginal zone B cells mimicked these parameters, suggesting both cell types contribute to a common protective mechanism. Our findings suggest that splenic neutrophils play a role in the protection against S. pneumoniae infection. Future studies will determine the contribution of resident vs recruited neutrophils to this protection and the role of these populations in mediating S.pneumoniae specific antibody production.
Plasmacytoid dendritic cell-derived IFNα modulates Th17 differentiation during Bordetella pertussis infection in mice (INC4P.345)
Authors: Smith andrew , Wu Victoria , Segal-Abramson Tzvia 
Published: January 2023

Abstract
Whooping cough is a highly contagious respiratory disease caused by Bordetella pertussis (B. pertussis). Recent studies have revealed a central role for Th17 cells in the resolution of the infection. Emerging studies document that type I interferons (IFN) suppress Th17 differentiation and IL-17 responses in models of infection and chronic inflammation. Plasmacytoid dendritic cells (pDCs) are a major source of type I IFNs. Therefore, we hypothesize that during B. pertussis infection in mice, pDC derived IFNa inhibits a rapid increase in Th17 cells. We found that IFNa-secreting pDCs emerge in the lungs in the early stages of infection, while a robust rise of Th17 cells in the lungs is detected at 15 days post-infection (dpi) or later. The presence of IFNa led to ablation of Th17 differentiation and proliferation in vitro. Furthermore, blocking IFNa produced by pDCs, in vivo prior to B. pertussis infection resulted in increase of Th17 frequency, inflammation and reduced bacterial loads in the airways of infected mice. Altogether, this work provides evidence of an inhibitory role for pDCs and pDC-derived IFNa in modulating Th17 responses during the early stages of B. pertussis infection, which may explain the prolonged nature of whooping cough.
Chronic STING-dependent signaling in phagocytes aggravates TREX1-contingent inflammatory disease (INM2P.353)
Authors: Ahn Jeonghyun 
Published: January 2023

Abstract
Patients defective in the 3’ repair exonuclease 1 (Trex1) suffer from Aicardi-Goutieres Syndrome (AGS) characterized by lethal encephalitis caused by high levels of circulating type I IFN production. Trex1-/- mice similarly suffer a short life span due to inflammatory myocarditis. Our data indicates that Trex1 is responsible for eliminating DNA species that otherwise activate STING (Stimulator of Interferon Genes), a key innate immune regulator responsible for facilitating cytosolic DNA-mediated cytokine production. Loss of STING (Trex1-/- STING-/-) was observed to dramatically extend the lifespan of Trex1-/- mice due to the prevention pro-inflammatory gene induction. However, we noted that Trex1-/- macrophages did not exhibit an augmented ability to produce cytokines following exposure to STING-dependent activators, but rather appeared chronically activated, exhibiting high basal level cytokine production. The source of STING activator was found to involve intrinsic genomic DNA left over from hematopoetic cell division processes that leaked into the cytoplasm, the elimination of which usually require Trex1. Accordingly, Trex1-manifested disease could be alleviated following adoptive transfer of normal bone marrow into Trex1-/- mice. Collectively, our data indicates that STING is responsible for pro-inflammatory gene induction induced by intrinsic self-DNA in Trex1 deficient mice and provides new insight into the molecular mechanisms underlining self-DNA-mediated disease.
Blockade of either SYK or BTK signaling pathways inhibits microparticle-induced type 2 inflammatory response (IRM11P.628)
Authors: Mishra Pankaj , Palma Mark , Millman Ariel , Hallab Nadim , Beebe Kathleen , Benevenia Joseph , Gause William 
Published: January 2023

Abstract
Wear debris microparticle release associated with total joint arthroplasty may contribute to harmful inflammation, osteolysis and implant failure. We examined the role of different molecular signaling pathways in microparticle-mediated inflammation. Intraperitoneal inoculation of cobalt chrome (CoCr) in mice induced a robust innate type 2 response with increases in Th2 cytokines, M2 macrophages, eosinophils and neutrophils compared to controls given vehicle only. Furthermore, this type 2 innate particle-induced response was independent of MYD88/TRIF and partially dependent on Caspase-1 signaling pathways. Blockade of the PGE-2 signaling pathway by administration of Aspirin or Celecoxib prior to CoCr inoculation caused a significant reduction in both neutrophils and macrophages with no effect on eosinophils and Th2 cytokines compared to the mice treated with CoCr + vehicle. Blockade of the SYK signaling pathway with the inhibitor BAY61-3606 or the BTK signaling pathway with Ibrutinib completely abrogated the CoCr-mediated innate inflammation and Th2 cytokines. These studies indicate that the type 2 immune response caused by solid particles is partially modulated by Caspase-1 and PGE-2 pathways and largely abrogated by blockade of upstream SYK and BTK signaling pathways. These studies provide potential targets for controlling the wear debris-mediated inflammation in synovial tissues that can contribute to aseptic loosening and implant failure.
C/EBPβ controls mucosal fungal immunity through regulation of β-defensins (IRM11P.630)
Authors: Simpson-Abelson Michelle , Childs Erin , Bishu Shrinivas , Solis Norma , Uematsu Satoshi , Conti Heather , Akira Shizuo , Filler Scott , Gaffen Sarah 
Published: January 2023

Abstract
Th17 cells and IL-17 play essential host-defensive roles in immunity to pathogens, particularly the commensal microbe Candida albicans. Although best known for activation of NF-κB, IL-17 potently activates CCAAT enhancer binding protein transcription factors, which are required for induction of many IL-17 target genes. C/EBPβ is translated into 3 distinct protein isoforms whose expression is regulated by the differential use of different translational initiation start sites, known as LAP* (38kDa), LAP (35kDa) and LIP (22kDa). We have shown that IL-17 induces the alternative translation of C/EBPβ, inducing a marked increase in LAP* and LIP. However, the functional significance of differential C/EBPβ isoform expression remains enigmatic. To determine the biological significance of C/EBPβ alternative translation, we used C/EBPβ-/- mice and a knockin that cannot generate the LAP isoform (C/EBPβM20A). We then assessed susceptibility of these mice to oral C. albicans infection. We found that C/EBPβ-/- but not C/EBPβM20 mice were highly susceptible to disease indicating that LAP is dispensable but C/EBPβ is required. In evaluating a panel of downstream genes to elucidate mechanisms of C/EBPβ-dependent immunity, the gene encoding β-defensin 3 (BD3) was the only IL-17 target gene strongly downregulated in susceptible C/EBPβ-/- mice. These results highlight a key role for C/EBP in antifungal immunity mediated by IL-17 and a previously unrecognized role for C/EBPβ in BD3 expression.
Immune modulatory mechanisms of lanthionine synthetase C-like protein 2 (IRM12P.656)
Authors: Washington Philipson Casandra , Heltzel Chase , Eden Kristin , Hontecillas Raquel , Bassaganya-Riera Josep 
Published: January 2023

Abstract
Lanthionine synthetase C-like protein 2 (LANCL2) has emerged as a novel therapeutic target for metabolic, infectious and immune-mediated diseases. We previously demonstrated that directly targeting LANCL2 through treatment with abscisic acid or 61610 results in efficacious anti-inflammatory responses during DSS-induced colitis however molecular mechanisms underlying this immunomodulatory function remain elusive. Here we report for the first time the use of LANCL2-deficient mice to investigate the mechanism of action and cell specificity of the protein. At the cellular level, bone marrow derived macrophages (BMDM), RAW cells, and gastric epithelial cells express comparable levels of basal LANCL2 mRNA. LPS-mediated inflammation in BMDM increased both mRNA and protein levels of LANCL2 that peaked 6 hours post-challenge and subsequently returned to normal levels after 12 hours. Similarly, DSS-induced colitis induced expression of LANCL2 in the colons of wild-type (WT) mice and correlated with elevated levels of proinflammatory cytokines. Interestingly, the absence of LANCL2 did not cause significant alterations in infiltrating cellular phenotypes however LANCL2-deficiency permitted exaggerated IL17 and IFNγ expression in colonic tissue when compared to WT counterparts at day 7 post-challenge. In conclusion, our findings indicate that LANCL2 is a novel regulator of inflammation that functions predominantly at the molecular level to down-regulate excessive inflammatory responses.