bluuebunny/crossref_metadata_2025_split huggingface dataset:
VTP-43742 is a potent and selective RORγt blocker that demonstrates oral efficacy in a mouse model of autoimmunity through suppression of IL-17A production (THER7P.945)
Authors: McGeehan Gerard , Bukhtiyarov Yuri , Zhao Yi , Meng Shi , Noto Paul , Stadanlick Jason , Kruk Barbara , Hardy Andrew , Lipinski Kerri , Kandpal Geeta , Algayer Bethany , Guo Joan , Guo Rong , Marcus Andrew , Lotesta Stephen , Dong Chengguo , Fan Kristi , Jia Lanqi , Yuan Jing , Zheng Yajun , Zhuang Linghang , Dillard Lawrence , Claremon David , Gregg Richard , Lala Deepak 
Published: January 2023

Abstract
Increased production of the inflammatory cytokine IL-17A by Th17 cells is a driver of multiple autoimmune disorders, including psoriasis, ankylosing spondylitis (AS) and psoriatic arthritis (PsA). The nuclear receptor RORγt, IL-23 and TGFb are required for the differentiation of Th17 cells. RORγt stabilizes the Th17 phenotype by increasing the expression of IL-23R and inducing the synthesis of IL-17A in Th17 cells. Antibodies targeting IL-23 or IL-17A are highly effective in the treatment of psoriasis, AS and PsA, validating the RORγt /Th17 pathway in human disease. VTP-43742 is an orally active inhibitor of RORgt that is being pursued for the treatment of autoimmune disorders. VTP-43742 binds to RORγt with high affinity (Ki=3.5 nM) and exhibits &gt;1000-fold selectivity versus the RORa and RORβ isotypes. VTP-43742 inhibits Th17 differentiation and IL-17A secretion from mouse splenocytes (IC50=57 nM) without affecting Th1, Th2, or Treg cell differentiation. In the MOG35-55/CFA immunized mouse EAE model, orally dosed VTP-43742 significantly suppressed clinical symptoms, demyelination and mRNA expression of multiple inflammatory markers in the spinal cord. Importantly, VTP-43742 inhibits the secretion of IL-17A from activated hPBMCs (IC50=18 nM) and human whole blood (IC50=192 nM) from healthy and psoriatic donors. Further, VTP-43742 is well absorbed after oral administration in multiple animal species and has pharmacokinetics consistent with once-a-day dosing in humans.
Lymphoma-associated mutant Herpesvirus entry mediator selects for B and T lymphocyte attenuator inhibitory signaling (TUM9P.1009)
Authors: Sedy John , Ware Brian , Schleimer William , Norris Paula , Ware Carl 
Published: January 2023

Abstract
Innate and adaptive lymphocytes respond to and manipulate their microenvironment through diverse cellular communication pathways including tumor necrosis factor (TNF) network signaling. A frequent target of cancers and viruses is the TNF receptor herpesvirus entry mediator (HVEM, TNFRSF14) and its ligand-receptor network because of the central role of this signaling system in maintaining immune homeostasis, and in regulating inflammatory responses. Notably, the biophysical basis for altered agonistic activity by pathogen and tumor-associated receptor variants remains unclear. Here we show that HVEM mutations associated with human follicular and diffuse large B cell lymphoma uniformly disrupt binding to CD160, while subsets of mutations retain binding to B and T lymphocyte attenuator (BTLA) and LIGHT (TNFSF14). We show that while HVEM activation of the inhibitory receptor BTLA is restricted by the expression of CD160 in lymphocytes, selectivity for BTLA by mutated HVEM or a viral paralog results in unhindered inhibition of ERK signaling and activation of IL-2 transcription. Together, the acquisition of ligand selectivity for BTLA by tumors and viruses illustrates a common mechanism of immune modulation utilized by intracellular pathogens to thwart inflammatory signaling.
Identification of a novel interleukin-10-producing B cell subset that fosters human cancer progression via programmed cell death-1 signals (TUM9P.1017)
Authors: Xiao Xiao , Kuang Dong-Ming 
Published: January 2023

Abstract
Regulatory B cells that are functionally defined by their ability to produce IL-10 downregulate inflammation and control T cell immunity. Here we identified a protumorigenic subset of B cells that constitutively expressed higher levels of programmed cell death-1 (PD-1) and constituted ~10% of all B cells in advanced stage hepatocellular carcinoma. These PD-1high B cells exhibited a unique CD5highCD24−/+CD27highCD38dim phenotype different from the phenotype of conventional regulatory B cells. TLR4-mediated Bcl-6 upregulation was crucial for PD-1high B cell generation in tumors, and that effect was abolished by IL-4-elicited STAT6 phosphorylation. Importantly, upon encountering PD-L1+ cells or undergoing PD-1 triggering, PD-1high B cells acquired regulatory functions that suppressed tumor-specific T cell immunity and promoted cancer growth via IL-10 signals. Our findings identify a novel subset of regulatory B cells in humans that contribute to immune privilege and cancer progression through PD-1-PD-L1 interactions.
Time-lapse imaging of contact dynamics between natural killer cells and cancer cells (TUM10P.1035)
Authors: Kim Jisung , Kim Yong Guk , Park Eun Jae , Kim Bo Yeong , Hong Jin Tae , Kim Youngsoo , Han Sang-Bae 
Published: January 2023

Abstract
Natural killer (NK) cell -mediated killing of cancer cells is essentially dependent on the direct cell-cell interaction. Using time-lapse imaging, we examine how NK cells contact melanoma cells at the single cell level. First, we analyzed contact dynamics of NK cells. Among 1,134 NK cells appeared in 36 movies in time-lapse imaging for 6 h, 121 NK cells (named competent NK, cNK) showed lytic contact with melanoma cells and 158 NK cells (named incompetent NK, iNK) showed non-lytic contact. Although contact numbers of both cell types was similar each other, the duration of contact appeared to be different. cNK cells contacted melanoma cells for 25 min without movement, but iNK cells contacted target cells for 13~14 min with moving around cancer cells. Both cell types moved at average speed of 5~6 um/min before contact and 1.6~2.1 um/min during contact. Next, we analyzed lytic process of melanoma cells, which could be described by two phases: rounding and membrane damage. Immediately after contacting cNK cells, all melanoma cells rounded up and it took 35 min (6-120) having small deviation. Then, melanoma cells lost membrane integrity and it took 103 (6-298) showing big deviation. Together, our data demonstrate that a small fraction of NK cells are cytotoxic to melanoma cells.
OVA-Texo/4-1BBL helped restore exhausted OVA-specific CD8+ T cell function during Adenovirus induced chronic infection. (VAC12P.1115)
Authors: Wang Rong , Xiang Jim 
Published: January 2023

Abstract
CD8+ T cell functional exhaustion has been adequately studied during established persistent viral infections such as LCMV in mice models as well as HIV and hepatitis C virus (HCV) in human chronic infections. Recombinant Adenovirus infection has also been reported to cause CD8+ T cell dysfunction and deletion. In this study, we established an Adenovirus induced chronic infection in C57BL/6 mice, characteristic by a long persistence of high percentage of antigen-specific CD8+ T cells, with increased inhibitory markers expression (PD-L1, PD-1, LAG-3, CD43) and compromised effector functions. Exosome-targeted CD8+ T cell vaccine is constructed through ConA stimulated non-specific CD8+T cells uptaking OVA pulsed DC-derived exosome (EXO), acting as CD8+Tc antigen presenting cells (Th-APCs), and capable of stimulating DC- and CD4+T cell-independent OVA-specific CTL responses leading to long-term immunity. We demonstrated that OVA-Texo/4-1BBL vaccine is able to provoke a successful immune response in chronic infected mice, increasing OVA-specific CD8+ cell level massively by approximately 3-fold, the primed CTLs could differentiate into functional CD8+ memory T cells. OVA-Texo/4-1BBL also helped restore exhausted CD8+ T cell function in the absence of CD4+ T cells. A sufficient increase of IFN-r production CD8+ T cell was measured after OVA-Texo/4-1BBL immunization compared with mice without treatment.
Virus reactivation is driven by inflammatory monocytes that are controlled by T cells (VIR9P.1163)
Authors: Chandran Ramakrishna , Cantin Edourad 
Published: January 2023

Abstract
The current dogma is that episodes of transient immunodeficiency triggered by various stressors reactivate herpesviruses that are ubiquitous in the human population. We established a novel model of Herpes simplex virus 1 (HSV) latency in immunodeficient B6.Rag mice inoculated with either a low (LD) or high dose (HD) inoculum by using IVIG and acyclovir to suppress virus replication and promote latency. Remarkably, heat stress induced in vivo reactivation (IVR) of latent HD-Rag mice resulted in uncontrolled HSV replication and fatal encephalitis (HSE) in all mice compared to transient reactivation in LD-Rag mice that all survived. The major difference between the two groups was the massive invasion of inflammatory Ly6Chigh monocytes (IM) into the brainstem (BS) of latent HD-Rag but not LD-Rag mice. T cell transfer prevented IVR induced fatal HSE in HD-Rag recipients by reducing inflammation, especially IM invasion of the BS. Surprisingly, IVR of lethally irradiated latent HD-Rag and wildtype B6 mice induced only transient reactivation resulting in 100% survival; notably, IM were absent in the BS of these mice. Thus, counter intuitively, our data show that IM resident at sites of latency promote rather than suppress HSV reactivation and by modulating IM invasion into the BS, T cells control virus reactivation. This is a novel finding with important clinical implications, especially for treatment of neonates and immunosuppressed cancer patients undergoing transplant therapy.
CD4+ FoxP3+ regulatory T cells control effector T cell responses to murine cytomegalovirus (VIR10P.1169)
Authors: almanan maha , Raynor Jana , wang Mei , Chougnet Claire , cardin Rhonda , Hildeman David 
Published: January 2023

Abstract
Human cytomegalovirus causes a persistent, lifelong infection. The virus persists in a latent state and undergoes intermittent subclinical viral reactivation. Such reactivation drives the accumulation of “inflationary” T cells that suppress viral replication. While T cells are critical to maintain control of infection, the factors that prevent T cells from driving complete viral elimination remain unclear. Here, we investigated the role of regulatory T cells (Treg) in a mouse model of persistent CMV infection using FoxP3-diphtheria toxin receptor (DTR) mice. FoxP3-DTR mice were infected with murine CMV (MCMV) and then Treg were depleted via administration of DT several months later. Treg depletion resulted in a dramatic increase in the numbers of MCMV-specific CD4 and CD8 T cells in the spleen which functionally, produced significantly higher levels of IFN-γ and IL-2. Strikingly, Treg depletion led to a significant decrease in splenic viral load. We are initiating experiments using a spleen viral reactivation assay, to determine the effect of Treg depletion on latent virus. Combined, these data suggest that Treg promote persistent MCMV infection, in part, by suppressing T cell activation.
Localization of iNKT subsets determines lipid response (APP2P.101)
Authors: Wang Haiguang , Lee You Jeong , Hogquist Kristin 
Published: January 2023

Abstract
iNKT cells can be rapidly activated by α-galactosylceramide (α-GalCer), and α-GalCer or analogues of it are being tested for clinical use against certain cancers and autoimmune diseases. Recent studies in our lab showed that iNKT cells are comprised of NKT1, NKT2 and NKT17 effector subsets. In the current study, we asked whether these different iNKT subsets respond equivalently to α-GalCer using Nur77GFP transgenic mice to quantify the antigen specific response. We observed a robust response by NKT1, but not NKT2, in the spleen, despite in vitro stimulation showing they could be equivalently activated. Using an in vivo antibody-labeling technique, we found that NKT1 possess significantly higher accessibility to blood than NKT2, suggesting that NKT1 cells are preferentially localized to red pulp. This was confirmed by tetramer-based immunofluorescence studies. As expected, i.v. labeled iNKT cells showed a greater response than unlabeled cells. Thus, iNKT cells with accessibility to blood are the predominant cells to respond to lipid. Consistent with this, we observed a robust response of liver iNKT cells, and no response by lymph node iNKT cells, regardless of the subset. Because iNKT subsets produce distinct cytokines, their differential response to i.v. lipids is a critical consideration in therapeutic applications.
Increased PKC signaling in lupus La reactive B cells promotes development of the transitional T3 population and memory B cells in autoimmune BXD2 mice (BA3P.108)
Authors: Hamilton Jennie , Li Jun , Wu Qi , Yang PingAr , Luo Bao , Li Hao , Bradley John , Taylor Justin , Randall Troy , Mountz John , Hsu Hui-Chen 
Published: January 2023

Abstract
We recently identified that lupus prone BXD2 mice exhibited defective clearance of apoptotic debris. In vivo administration of excessive autologous apoptotic cells lead to elevation of IgG anti-La13-27 responses in BXD2 mice but not normal B6 mice, suggesting chronic self-antigen stimulation plus B-cell tolerance defects may be related to the IgG anti-La13-27 responses. To interrogate if there was abnormal maturation of anti-La13-27 B cells in naïve BXD2 mice, a recently developed La13-27 tetramer was used with CD80 and PD-L2 co-staining to reveal that there was a significantly higher frequency of memory CD80+PD-L2+ La13-27+ B cells in BXD2 (17%), compared to that in B6 (3%) mouse spleens. Tetramer analysis of CD93+ transitional B cells showed that there was an abnormal skewing of the La13-27+ T3 population, but not T1 or T2 B cells, in BXD2 (40%) compared to that in B6 (18%) mouse spleens. Survival of anti-La13-27 producing B cells in BXD2 but not B6 mice was associated with increased protein kinase C (PKC) activation, as stimulation of B cells with PMA+ionomycin which promotes survival of BCR stimulated transitional B cells resulted in an increase in IgG. As T3 B cells are considered anergic B cells maintaining self-tolerance through rapid turnover in vivo, the abnormal expansion of these B cells coupled with increased BCR signaling response in BXD2 mice may present a risk for B-cell tolerance loss to La13-27, leading to maturation into IgG secreting cells.
Lupus risk-variant increases pSTAT1 binding and decreases ETS1 expression (BA4P.137)
Authors: Lu Xiaoming , Zoller Erin , Weirauch Matthew , Wu Zhiguo , Namjou-Khales Bahram , Shen Nan , Greis Kenneth , Kaufman Kenneth , SLEGEN The , Harley John , Kottyan Leah 
Published: January 2023

Abstract
Genetic variants near gene ETS1 have been associated with Systemic Lupus Erythematosus (SLE) in independent cohorts of Asian ancestry. ETS1 is a critical driver of immune cell function and differentiation, and mice deficient in ETS1 develop an SLE-like autoimmunity. We performed a fine-mapping study of 14,551 subjects using multi-ancestral cohorts, starting with genotyped variants and imputing to all common variants spanning the ETS1 locus. By constructing genetic models using frequentist and Bayesian association methods, we identified a set of 16 variants that are statistically most likely to be causal. We functionally assessed each of these variants based on their biological function predictions. Of the four variants that we experimentally examined, only rs6590330 differentially binds lysate from B cells. Using mass spectrometry, we found increased binding of the transcription factor signal transducer and activator of transcription 1 (STAT1) to DNA near the risk allele of rs6590330 compared to the non-risk allele. Western blot analysis and chromatin immunoprecipitation of pSTAT1 in B cells heterozygous for rs6590330 confirmed that the risk allele increased binding to the active form of STAT1. eQTL analysis indicates that the risk allele of rs6590330 is only associated with decreased ETS1 expression in Han Chinese. We propose a model in which the risk allele of rs6590330 increases SLE risk by enhancing the binding of pSTAT1, resulting in repression of the ETS1 expression.
Alleviation of joint inflammation by lymphatic pump treatment of rats with adjuvant-induced arthritis (CAM1P.163)
Authors: Volin Michael , Zanotti Brian , Larimer Kaitlin , Gustafson Nicholas , Shah Pratik 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is characterized by synovial inflammation, cytokine production and lymph node collapse. Lymphatic pump treatment (LPT) is an osteopathic manipulative treatment used to reduce edema through increasing lymphatic circulation. This study examined the therapeutic potential of LPT on rats with adjuvant-induced arthritis (AIA). We hypothesize that LPT would lessen joint inflammation through increasing lymphatic drainage resulting in the reduction of cytokines in synovial space. AIA was induced in female Lewis rats and randomly placed into a LPT groups or sham groups. Experiments were repeated using different LPT protocols varying the frequency and number of LPT treatments. However in each experiment LPT consisted of rhythmically pressing below the rib cage once a second for 30 seconds, while the sham group was held for 30 seconds. Joint inflammation was determined by measuring ankle circumferences and by articular index scoring. Ankles were collected and homogenized to determine changes in synovial cytokine levels by magnetic immunoassays and ELISA. Clustering LPT treatments three-times a day at the peak of joint inflammation had the greatest impact on arthritis. Animals treated with LPT in this manner exhibited significantly reduced ankle edema and inflammation. Additionally, synovial levels of inflammatory cytokines (GRO, IL17, and IL-6) were reduced with LPT. These results indicate that LPT can reduce joint edema and inflammation in a rat model of RA.
A computational model for mechanistic investigation of negative feedback in interleukin-17 receptor signaling (CCR3P.201)
Authors: Sheehan Robert , Garg Abhishek , Gaffen Sarah , Faeder James 
Published: January 2023

Abstract
IL-17 is a pro-inflammatory cytokine that promotes autoimmunity and protects against certain pathogens. Here, we present a computational model of signaling downstream of the IL-17 receptor. To manage the complexity of the system we use a rule-based modeling approach, in which signaling proteins are modeled as structured objects and rules describe their biochemical interactions, allowing us to consider all possible complexes and phosphoforms generated from a set of basic molecules. This model encompasses major signaling components downstream of IL-17R, which activate NF-κB and in turn promote the production of pro-inflammatory cytokines. NF-κB also promotes production of A20, a deubiquitinating enzyme that inhibits NF-κB activation in multiple settings. We recently showed that A20 acts as a negative regulator of the IL-17 signaling pathway. By modeling A20 inhibition of TRAF6 and IKK, we are able to recapitulate experimentally-observed oscillations in NF-κB activity and A20 expression. Novel experimental data identifying the dynamics of signaling intermediates has further constrained model behavior and informed model development. Modeling the depletion of IκB, followed by its rebound above baseline levels, required the addition of a stochastic model of transcription factor binding, paired with a deterministic model of signaling events. We are currently using the hybrid model to identify which of several potential mechanisms A20 uses to attenuate IL-17 signaling.
The introduction of RNA-DNA differences underlies inter-individual variation in the human IL12RB1 mRNA repertoire (CCR3P.204)
Authors: Robinson Richard 
Published: January 2023

Abstract
IL12 and IL23 promote numerous aspects of human immunity, including autoimmunity and mycobacterial disease resistance. IL12RB1 encodes human IL12Rβ1, a surface receptor that signals in complex with IL12Rβ2/IL23R to relay the pro-inflammatory signals of IL12/IL23. Here, we present data demonstrating that in primary human T-lymphocytes, sequence diversity reminiscent of TCRA/TCRB diversity is introduced into IL12RB1 mRNAs via RNA DNA differences (RDDs). RDDs are ribonucleotides present in an mRNA that are not genome encoded; as such they are distinct from genomic single nucleotide polymorphisms. First observed in protozoa, RDD-introduction is now known to be a mechanism employed by many species to suppress protein function. In the case of IL12RB1, we show that in T-cells of healthy individuals the majority of IL12RB1 mRNAs expressed contain RDDs that concentrate in exons encoding IL12Rβ1’s cytokine binding region. IL12RB1 RDDs comprise multiple mutation types, are detectable by both Ion Torrent sequencing and classic Sanger sequencing, and decrease in frequency following T-cell activation. As a result of RDD-introduction, IL12Rβ1 proteins have an altered amino acid sequence that could not be predicted based on genome sequencing alone. While the underlying mechanisms of sequence diversification differ for TCRA/TCRB and IL12RB1, the consequence of this diversification is the same - the expression of proteins of variable amino acid sequence and variable immunological function.
Chromodomain helicase DNA-binding 4 is required for proliferation, distal VH rearrangements and developmental progression of B cell progenitors (HEM1P.221)
Authors: Hagman James , Dege Carissa , Straign Desiree , Jia Haiqun , Walton Kendra , Lukin Kara , Lei Hong , Danhorn Thomas , Feeney Ann 
Published: January 2023

Abstract
Chromodomain Helicase DNA-binding protein 4 (CHD4, or Mi-2β) is a catalytic core subunit of Nucleosome Remodeling and Deacetylase (NuRD) complexes, which regulate chromatin structure and transcription in lymphocytes. CHD4 activities include ATP-dependent mobilization of nucleosomes, DNA binding and binding to histone tails. Here, we investigated requirements for CHD4 in B cell development in a mouse model system. We utilized Chd4flox/flox:Cd79a-Cre (Chd4 cko) mice, which inactivate Chd4 genes selectively in early B cell progenitors. These mice confirmed that CHD4 is essential for B lymphopoiesis. Following the loss of CHD4 expression, B cell development is arrested at the pro-B cell stage. Peripheral B220+ cells were nearly absent. To address the basis of the observed developmental arrest, we measured effects of the lack of CHD4 on proliferation and Igh gene rearrangements. CHD4-deficient pro-B cells fail to proliferate in response to IL-7. Furthermore, pro-B cells lacking CHD4 complete proximal VH to DJH rearrangements, but rearrange distal VH segments only rarely. Overall, our data demonstrate that CHD4 and NuRD complexes are essential for multiple aspects of early B cell development, including V(D)J recombination, proliferation and survival of pro-B cells.