bluuebunny/crossref_metadata_2025_split huggingface dataset:
A transcription factor network in Th9 cells (P1229)
Authors: Jabeen Rukhsana , Goswami Ritobrata , Awe Olufolakemi , Kaplan Mark 
Published: January 2023

Abstract
Th9 cells are the most recent addition to the spectrum of T helper cell subsets that differentiate in the presence of TGF-β and IL-4. Th9 cells promote allergic inflammation, anti-tumor immunity, and may contribute to the regulation of auto inflammatory disease. Whether Th9 cells express a characteristic transcriptional signature has not been determined. To define the identity of Th9 cells, we performed a microarray analysis and identified 629 genes enriched in Th9 cells compared to Th2 and Treg cells. Of these genes, we chose a panel of 14 genes based on potential relatedness to Th9 function, and confirmed Th9-enriched expression in a larger panel of T helper subsets using quantitative PCR. The Th9 enriched genes encoded transcription factors including Erg, Maf, Ahr and Crem, and surface receptors including Il17rb, Ccr4 and Ccr8. Using mice deficient for the transcription factors that promote Th9 development including STAT6, IRF4 and PU.1, we distinguished the impact of each factor on this subset of genes. Thus our data indicates that Th9 cells have an identifiable transcriptional signature, and we have defined a transcription factor network that regulates Th9 gene expression.
Distinct dendritic cell subsets dictate the fate decision between effector and memory CD8+ T cell differentiation (P6172)
Authors: Kim Taeg , Gorski Stacey , Hahn Steven , Braciale Thomas 
Published: January 2023

Abstract
An effective host response to a microbial pathogen requires both the generation of effector T cells as well as formation of durable T cell memory. Dendritic cells (DCs) play a central role in initiating the adaptive T cell response to microbial infection, and multiple distinct DC subsets have the potential to take up and present microbial antigen. However, the contribution of different DC subsets to the generation of effector and memory CD8+ T cells in vivo following infection and the DC dependent mechanisms to account for this dichotomy remain incompletely understood. Here we show that the two major migratory respiratory DC (RDC) subsets responding in the lung draining lymph nodes to influenza virus infection, CD103+ and CD11bhi RDC, activate naïve virus-specific CD8+ T cells along distinctly different differentiation pathways. CD103+ RDC in vivo stimulate the generation of activated CD8+ T cells with features characteristic of CD8+ T effectors that is activated T cells which efficiently leave the lymph nodes and home to the infected respiratory tract. In contrast, stimulation of naïve virus-specific CD8+ T cells by CD11bhi RDC give rise to CD8+ T effectors which are preferentially retained within the draining lymph nodes during the acute phase of infection, and which exhibits properties of central memory CD8+ cells. DC-dependent cellular and molecular basis for refining this dichotomy for in vivo CD8+ T cell differentiation are further delineated.
Kruppel-like factor 6 and inducible nitric oxide synthase regulates apoptosis during respiratory syncytial virus infection (P6198)
Authors: Segovia Jesus , Mgbemena Victoria , Chang Te-Hung , Bose Santanu 
Published: January 2023

Abstract
Human respiratory syncytial virus is a highly pathogenic lung-tropic virus that causes severe respiratory diseases such as pneumonia and bronchiolitis in infants, children and the elderly. Various innate immune mediators including nitric oxide are produced in response to respiratory syncytial virus infection and play a critical role in host defense. Generation of nitric oxide during respiratory syncytial virus infection is dependent on inducible nitric oxide synthase expression. Although nitric oxide plays a crucial role in combating pathogen invasion and spread, it can also lead to an exaggerated inflammatory response and tissue damage. Nitric oxide-mediated apoptosis is responsible for exacerbated disease pathology during respiratory syncytial virus infection, however the mechanism is unknown. In the current study we have identified kruppel-like factor 6 as a critical transcription factor required for inducible nitric oxide synthase gene expression during respiratory syncytial virus infection. Furthermore, we have also uncovered the role of inducible nitric oxide synthase as a critical host factor regulating apoptosis during respiratory syncytial virus infection.
Interferon lambda confers antiviral protection on the epithelium (P6186)
Authors: Mahlakoiv Tanel , Staeheli Peter 
Published: January 2023

Abstract
Type III interferon (IFN-λ) belongs functionally to the interferon family, although it is structurally related to IL-10 family of cytokines. We have used a reporter mouse lacking type I IFN receptor to identify IFN-λ-responsive cells in vivo by staining tissue sections with antibodies that recognize the IFN-induced Mx1 protein. We found that epithelial cells lining the respiratory, gastrointestinal and reproductive tracts are responding most strongly to IFN-λ stimulation. Additionally, IFN-λ-induced Mx1 was detectable in epithelial cells of the skin, placenta, fetal membranes, ovaries, mammory glands, cornea and conjunctiva. In vitro experiments on various epithelial cell lines not only confirmed this finding, but also demonstrated the potent antiviral activity of exogenous IFN-λ against a diverse set of viruses. Using knockout mice lacking either functional type I or type III IFN receptors, we demonstrated that IFN-λ is redundant to type I IFN functions in the lung upon infection with various respiratory viruses, whereas a predominant protection against rotavirus infection of the small intestinal epithelium was mediated by IFN-λ. Interestingly, mice lacking functional receptors for IFN-λ showed higher susceptibility not only to rotavirus, but also to another enteric virus, the reovirus. Taken together, a picture emerges showing that IFN-λ is a potent antiviral cytokine mediating protection against viral pathogens which infect the epithelium.
Transgenic expression of survivin compensates the OX40-deficiency in driving Th2 development and allergic inflammation (P1184)
Authors: Song Jim , Lei Dylan , Haque Rizwanul 
Published: January 2023

Abstract
The inhibitor of apoptosis family molecule survivin has been suggested to be a crucial intermediate in signaling pathways leading to T cell development, proliferation, and expansion. However, the importance of survivin to T cell-driven inflammatory responses has not been demonstrated. Here, we show that survivin transgenic mice exhibit increased antigen-driven Th2 lung inflammation, and that constitutive expression of survivin reverses defective lung inflammation in the absence of OX40 costimulation. We found that while OX40-deficient mice are compromised in generating Th2 cells, airway eosinophilia, and IgE responses, survivin transgenic and OX40-deficient mice generate normal Th2 responses and exhibit strong lung inflammation. These data suggest OX40 costimulation crucially engages survivin during antigen-mediated Th2 responses, and promote the notion that targeting survivin could be therapeutically useful for controlling allergic responses or lung inflammation.
Signaling through the P2X7R for ATP induces human cutaneous innate and adaptive Th17 responses. (P4110)
Authors: Mathers Alicia , Killeen Meaghan , Ferris Laura , Kupetsky Erine , Falo, Jr. Louis 
Published: January 2023

Abstract
Human cutaneous DCs have the ability to prime and bias Th17 lymphocytes. However, the factors that stimulate cutaneous DCs to induce Th17 responses are not well known. We hypothesize that alarmins, such as ATP, play a pivotal role in the induction and maintenance of cutaneous immune responses by stimulating DC maturation, chemotaxis, and secretion of IL-1β and IL-6, Th17 biasing cytokines. Utilizing an ex vivo human cutaneous model, we have determined that signaling purinergic receptors, predominantly P2X7R, via an ATP analog initiates innate proinflammatory inflammation, DC17 differentiation, and the subsequent induction of Th17 biased immunity. Moreover, our results suggest a potential role for P2X7R signaling in the initiation of psoriasis pathogenesis, a Th17 dependent autoimmune disease. In support of this, we observed the increased presence of P2X7R in non-lesional and lesional psoriatic skin compared to normal healthy tissues. Interestingly, there was also a P2X7R variant highly expressed in lesional psoriatic skin compared to non-lesional psoriatic and normal healthy skin. In conclusion, we have established that purinergic signaling in the skin induces innate inflammation leading to the differentiation of human Th17 responses, which have implications in the pathogenesis and potential treatment of psoriasis.
Proatherogenic conditions promote autoimmune Th17 responses in vivo (P4136)
Authors: Chung Yeonseok , Lim Hoyong , Kim Young Uk , Sun Hua , Hanabuchi Shino , Teng Babie 
Published: January 2023

Abstract
Although patients with atherosclerosis have a higher incidence of systemic autoimmune diseases, the relationship between proatherogenic factors and autoimmune T cell responses is poorly understood. Mice lacking both LDL receptor and apolipoprotein B mRNA editing enzyme (Ldlr-/-Apobec1-/-; LDb) are hyperlipidemic and prone to atherosclerosis. Here, we show that LDb mice exhibit increased IL-17 in circulation as well as in the aortic sinus area, which was attributed to a preferential enhancement of Th17 cells in the secondary lymphoid organs. In addition, the environment within LDb mice was substantially favorable for the Th17 polarization of auto-reactive CD4+ T cells during homeostatic proliferation. In vitro, the addition of oxidized LDL, but not native LDL, promoted dendritic cell-mediated Th17 polarization by triggering IL-6 and IL-1 in a MyD88-dependent fashion. Furthermore, myelin oligodendrocyte glycoprotein (MOG)-reactive CD4+ T cells expanded in the presence of oxidized LDL expressed increased levels of Th17 signature genes, and induced more profound experimental autoimmune encephalitomyelitis (EAE) when transferred into naïve mice. Our findings demonstrate that proatherogenic factors induce the polarization and functional maturation of autoimmune Th17 cells, which may be critical for the pathogenesis of atherosclerosis and other related autoimmune diseases.
Human CCR6+CXCR3+ “Th1/17” cells are polyfunctional and can develop from Th17 cells under the influence of IL-1β and IL-12 (P4122)
Authors: Duhen Thomas , Campbell Daniel 
Published: January 2023

Abstract
In humans, memory CD4+ T cells co-expressing the chemokine receptors CCR6 and CXCR3, and producing both IL-17 and IFN-γ, have been proposed to be highly pathogenic in several autoimmune disorders. However, their developmental pathways, relationship with “classic” Th17 and Th1 cells and physiological role in normal immune responses are not well understood. Here, we examined CCR6+CXCR3+ T cells from healthy individuals, and found that ex vivo those cells expressed higher levels of IL12Rβ2 and IL23R compared to Th17 and Th1 cells and displayed an enhanced response to IL-12 and IL-23. Interestingly, Th17 cells cultured in the presence of IL-1β upregulated IL-12Rβ2, and IL-1β together with IL-12 induced IL-17+ IFN-γ+ cells and strongly increased the expression of T-bet, IL-12Rβ2 and IL-23R without inhibiting RORγt. CCR6+CXCR3+ T cells shared antigen specificities with Th17 and Th1 cells, and T cells specific for extracellular bacteria such as S. pneumoniae, E. coli and L. rhamnosus were highly enriched in this subset. Furthermore, a significant proportion of CCR6+CXCR3+ T cells expressed the gut-homing molecule α4β7. Thus, CCR6+CXCR3+ CD4+ T cells are a functionally distinct population of T cells with broad antigen specificity and migratory potential that can develop from “classic” Th17 cells under the influence of IL-1β and IL-12. Further characterization of these cells will reveal new insights into CD4+ T cell differentiation and function in infection and autoimmunity.
Glucose metabolism in CD4+ T cell subsets modulates inflammatory disease and autoimmunity (P4126)
Authors: Gerriets Valerie , Macintyre Andrew , Nichols Amanda , Rathmell Jeff 
Published: January 2023

Abstract
Upon stimulation, CD4+ T cells differentiate into effector (Teff) or regulatory (Treg) T cells, the balance of which is crucial to prevent inflammation or autoimmunity. One approach to treat autoimmune disease is to modulate T cell differentiation to favor Treg over Teff cells. We examined the metabolite and metabolic gene expression profiles of the subsets and found that they have distinct metabolic requirements and programs. Teff have a high expression of the glucose transporter GLUT1 as well as glucose metabolism genes and a high rate of glycolysis. In contrast, Treg instead have high expression of mitochondrial metabolism genes and low GLUT1 levels. The effect of metabolic differences to modulate the Teff:Treg balance can be tested directly via genetic modulation of GLUT1. In vitro, T cells from GLUT1 transgenic mice are more inflammatory than wildtype littermates. Alternatively, T cells from GLUT1 knockout mice are unable to increase glucose metabolism upon activation and have fewer Th1, Th2 and Th17 cells. Despite less total CD4+ T cells, these mice have normal natural and inducible Treg numbers. In a mouse model of inflammatory bowel disease (IBD), deletion of GLUT1 in T cells inhibits IBD disease progression. The mice have less CD4+ T cells in the mesenteric lymph nodes and decreased inflammatory cytokine production than wildtype control T cells. Thus, the modulation of glucose metabolism may provide an alternative treatment for inflammatory and autoimmune disease.
TCR repertoire analysis of La/SS-B autoantigen-responsive peripheral blood CD4+ T cells using single-cell RT-PCR: a proof of principle case study (P3249)
Authors: Joachims Michelle , Pan Zijian , Moore Jacen , James Judith , Scofield Robert , Sivils Kathy , Thompson Linda , Farris A. 
Published: January 2023

Abstract
TCR repertoire analysis in a variety of biological settings has largely consisted of TCR spectratyping of V-beta expression or V-specific antibody staining. Here, we combine in vitro stimulation of CFSE-labeled antigen specific T cells with a single cell nested multiplex RT-PCR to efficiently capture the unique T cell repertoire of autoantigen La/SS-B specific CD4+ T cells in a systemic lupus erythematosus and Sjögren’s syndrome patient with precipitating levels of anti-La and Ro antibodies. CFSE-labeled peripheral blood cells were cultured with either purified La protein or a mixture of human La peptides, and CD4+CFSElow cells were single sorted into PCR plates. TCR alpha and beta chain cDNAs were amplified and sequenced. The efficiency of paired TCR chain recovery was 60-65%, and was similar between the populations analyzed. While the magnitude of proliferative responses to the two types of La antigen-specific stimuli were similar, the responding T-cells displayed little overlap in TCR sequence. The sequences derived from the peptide pool-stimulated CD4+ cells were pauci-clonal, with 2 clones dominating more than half the response (58/93 cells). Conversely, the purified protein-responsive CD4+ TCRs were distributed among multiple clonotypes. Our approach maps the T cell repertoire of human antigen responsive cells and may have great utility for elucidating the properties of a variety of antigen-specific T cells in diverse clinical settings.
T cell evaluation in cytotoxicity studies (P3271)
Authors: Gillis Katherine , Clor Julie , Khan Asima , Tyagarajan Kamala 
Published: January 2023

Abstract
A comprehensive mechanistic understanding of cytotoxic compounds action requires knowledge of their impact on different immune sub-populations such as CD4 and CD8 T cells. In this study, we explored the mechanism of action of Gambogic acid, a natural product with potent anti-tumor activity and Anisomycin, an antibiotic that inhibits protein synthesis on peripheral blood mononuclear cells (PBMCs) using assays that combine CD4 and CD8 T cell identification with cell health parameters followed by microcapilllary cytometry. Our studies demonstrate that gambogic acid results in significant apoptosis in both CD4 and CD8 T cells. Further, mitochondrial depolarization plays a significant role in gambogic acid induced cytotoxicity of both CD4 and CD8 T cells with a greater percentage of cells demonstrating loss of mitochondrial membrane potential at lower concentration compared to other apoptosis markers. Anisomycin also results in significant mitochondrial membrane potential loss in T cells. Significant Caspase activation was observed in PBMC’s on treatment with anisomycin; with Caspase 3/7, Caspase 8 and Caspase 9 activity being observed in CD4 and CD8 T cells. Data on dose and time based responses in the sequence of cell health impacts will be further presented for these compounds on PBMC. Evaluating mechanistic impact of compounds on T lymphocytes can provide a more enriched understanding of complete activity of compounds that induce cytotoxicity.
Immune activating effects of stimulation of TLR agonists and cytokines on keratinocytes from a patient with a CARD14 mediated pustular psoriasis (P6283)
Authors: Chen Yongqing , Liu Yin , Huang Yan , Yee Carole , McBride Alison , Morasso Maria , Bowcock Anne , Lowes Michelle , Goldbach-Mansky Raphaela 
Published: January 2023

Abstract
Caspase recruitment domain-containing protein 14 mutations are the first monogenic defect causing psoriasis and pityriasisi rubra pilaris (PRP), linking psoriasis and PRP into one disease spectrum. Interestingly the disease responds well to the treatment with anti-p40 antibody (Ustekinumab) blocking IL-12 and IL-23. The mutant protein is mainly expressed in keratinocytes, leading us to explore whether keratinocyte dysfunction triggers the inflammatory phenotype through the recruitment of hematopoietic cells to the skin. CAR14 mutant keratinocytes produced high level of IL-8 to Toll-like receptor 3, 5, 9, IL-1a/b, IL-36, and TNF stimulation than control cells. IL-17A synergizes with Poly IC, Flagellin, and TNF, but not with IL-12/23, suggesting that Ustekinumab may not directly block keratinocytes but indirectly through inhibit IL-17A produced from likely hematopoietic cells. Using siRNA knock down assay of CMB (CARD-MALT1-BCL10) complex, we show that IL-17A induced IL-8 production is MALT1-dependent; thus raising the question what is the role of CARD14 in IL-17 mediated signal. Our data suggests that while IL-12/23 does not activate keratinocytes, IL-17A is a potent activator and even synergistically with other cytokine stimulant from hematopoietic cells during skin inflammation.
Deubiquitinase A20 mediates feedback inhibition of IL-17 receptor signal transduction (P6267)
Authors: Garg Abhishek , Ahmed Mushtaq , Vallejo Abbe , Ma Averil , Gaffen Sarah 
Published: January 2023

Abstract
IL-17A (IL-17) is a proinflammatory cytokine that mediates host defense against extracellular pathogens but also contributes to the pathogenesis of various autoimmune diseases, such as rheumatoid arthritis (RA), multiple sclerosis (MS) and psoriasis. Despite some homology with the toll-like receptors, IL-17 receptor subunits (IL-17RA and IL-17RC) do not recruit adaptors such as MyD88, but instead associate with an E3 ubiquitin ligase Act1. Upon recruitment, Act1 ubiquitinates Traf6, resulting in the activation of the NF-κB and MAPK pathways. Details of the molecular events involved in negative regulation of IL-17 signaling still remain poorly understood, but are important clinically in understanding IL-17 mediated autoimmune pathology. Since both Act1 and Traf6 have ubiquitin ligase activity, deubiquitination could be an effective strategy of inhibiting IL-17 signaling. We focused on the enzyme A20 (Tnfaip3), as mutations in its gene locus are associated with many autoimmune diseases. Indeed, we found that absence of A20 results in increased IL-17-dependent proinflammatory gene expression and is associated with increased NF-κB activation. Unexpectedly, A20 associates with IL-17RA at its distal C-terminal domain, a domain that has been previously demonstrated to play a role in mediating negative regulation of IL-17 signaling. In summary, regulation of IL-17 signaling by A20 might provide an interesting mechanistic approach for inhibiting this pathway.
The adaptor protein CIKS/Act1 is essential for imiquimod-induced psoriasis. (P6269)
Authors: Ha Hye-Lin , Siebenlist Ulrich 
Published: January 2023

Abstract
Psoriasis is a skin disease characterized by abnormal keratinocyte (KC) proliferation and differentiation and by an influx of inflammatory immune cells. The IL-23/IL-17 axis is thought to play an important role in the pathogenesis of psoriasis. Imiquimod (IMQ) can induce/exacerbate psoriasis in patients and it induces a psoriasis-like condition in mice. In addition to IL-17A/F, IL-17C has also recently been implicated in IMQ-induced psoriasis. To determine by what mechanisms IL-17 cytokines critically contribute to psoriasis, we investigated a mouse model in which signaling by all IL-17 cytokines is abrogated due to the loss of the obligate adaptor protein CIKS/Act1. Importantly, the gene for CIKS has also been identified as a strong susceptibility locus in psoriasis. Here we show that IMQ induced many of the typical phenotypes of human psoriasis in wild-type mice, but largely failed to do so in CIKS-deficient mice. In addition to promoting the influx of inflammatory cells, IL-17 signaling via CIKS contributed to increased proliferation of KCs and inhibited their terminal differentiation. Mice lacking CIKS in KCs only were protected from some, but not all of the pathologic phenotypes associated with psoriasis in this model. These data demonstrate that CIKS is essential for IMQ-induced psoriasis by mediating the signaling of IL-17 cytokines in several distinct cell types. Thus our findings identify CIKS as a promising target for therapeutic intervention in this disease.
Follistatin-like protein 1 regulates IL-17 signalling by influencing the expression of IL-17 receptor A. (P6271)
Authors: Campfield Brian , Hirsch Raphael , Kolls Jay 
Published: January 2023

Abstract
Background: IL-17 is a critical proinflammatory cytokine in Collagen Induced Arthritis (CIA) and lung host defense. Follistatin-like protein-1 (FSTL-1), a proinflammatory glycoprotein mediates the development of CIA. FSTL-1 mediates IL-17-dependent stimulation of IL-6 by an unknown mechanism. Methods: ST2 cells with shRNA for FSTL-1 (shFSTL-1) and control (shCntrl) sequences were stimulated with 8ng/mL IL-17 and 2ng/mL TNFα. Transcript abundance was assessed by qRT-PCR. Protein secretion was assessed by Luminex. IL-17 receptor surface expression was assessed by FACS analysis. Results: Compared to shCntrl cells, shFSTL-1 cells had 1)decreased transcripts of IL-6 (275±3 vs. 43±3), G-CSF (1802±7 vs. 1325±93), IL-17RA (0.798±0.02 vs. 0.504±0.03) and IL-17RC (1.28±0.01vs. 0.72±0.1) relative to unstimulated cells, 2)decreased protein secretion IL-6 (25142 ± 229pg/ml vs. 2542 ± 37), G-CSF (388 ± 32 pg/ml vs. 27.8 ± 3.9 ) secretion, and 3)decreased IL-17RA surface expression when unstimulated (86.6% vs. 44.8%) or stimulated with ligand (14.1% vs. 5.2%) despite similar IL-17RC surface expression when unstimulated (93.6% vs. 91.3%) or stimulated (59.2% vs. 49.5%). Conclusions: FSTL-1 mediates IL-17 stimulated IL-6 and G-CSF production in ST2 cells. FSTL-1 influences transcript abundance of IL-17RA and IL-17RC, but shFSTL-1 attenuates surface expression of IL-17RA alone, suggesting that FSTL-1 regulates IL-17 signalling by influencing the cell surface expression of IL-17RA.