bluuebunny/crossref_metadata_2025_split huggingface dataset:
TERRA enhances production of senescence-associated inflammatory cytokines IL-6 and IL-8 (IRC5P.452)
Authors: Wang Pu 
Published: January 2023

Abstract
Aged or senescent cells develop a complex senescence-associated secretory phenotype (SASP). However, the mechanisms underlying the induction of the SASP are unclear. Telomeres are critical for genome stability and aging. Telomeres are transcribed into long non-coding telomeric repeat-containing RNA (TERRA) molecules. We demonstrate that TERRA and inflammatory cytokines were significantly elevated during senescence. Over expression of TERRA significantly upregulated IL-6 and IL-8, but IL-6 and IL-8 did not modulate TERRA expression. The results show that TERRA expression is related to senescence and increases production of SASP. Our findings expand the biological function of TERRA in senscence-assosciated inflammatory responses.
RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance (IRC5P.465)
Authors: Xiao Yanping , Yu Sanhong , Zhu Baogong , Bedoret Denis , Bu Xia , Francisco Loise , Hua Ping , Duke-Cohan Jonathan , Umetsu Dale , Sharpe Arlene , DeKruyff Rosemarie , Freeman Gordon 
Published: January 2023

Abstract
We report that programmed death ligand 2 (PD-L2), a known ligand of PD-1, also binds to repulsive guidance molecule b (RGMb) which was originally identified in the nervous system as a co-receptor for bone morphogenetic proteins (BMPs). PD-L2 and BMP-2/4 bind to distinct sites on RGMb. Normal resting lung interstitial macrophages and alveolar epithelial cells express high levels of RGMb mRNA, while lung dendritic cells express PD-L2. Blockade of the RGMb:PD-L2 interaction markedly impaired the development of respiratory tolerance by interfering with the initial T cell expansion required for respiratory tolerance. Experiments with PD-L2 deficient mice showed PD-L2 expression on non-T cells was critical for respiratory tolerance but expression on T cells was not required. Since PD-L2 binds to both PD-1, which inhibits anti-tumor immunity, and to RGMb, which regulates respiratory immunity, targeting the PD-L2 pathway has therapeutic potential for asthma, cancer and other immune-mediated disorders. Understanding this pathway may give insights into how to optimally modulate the PD-1 pathway in cancer immunotherapy while minimizing adverse events.
A commensal microbial molecule confers protective anti-inflammatory responses employing both innate and cognate interactions (IRC5P.468)
Authors: Dasgupta Suryasarathi , Kasper Dennis 
Published: January 2023

Abstract
The microbiota is critical in shaping the mammalian host’s immune system. Polysaccharide A, the archetypical immunomodulatory microbial molecule of the gut commensal Bacteroides fragilis, induces regulatory T cells to secrete the anti-inflammatory cytokine interleukin 10. We show, in a model of colitis, that PSA requires both innate and adaptive immunity to generate protection. Dendritic cells mediate PSA’s effect on IL-10 production. Unlike conventional DCs, plasmacytoid DCs exposed to PSA do not produce the proinflammatory cytokines tumor necrosis factor alpha and IL-12, but PDCs do specifically stimulate IL-10 secretion by CD4+ T cells and efficiently mediate PSA-mediated immunoprotection. PSA induces and preferentially ligates Toll-like receptor 2 on PDCs but not on CDCs. Compared with other TLR2 ligands, PSA better enhances PDC expression of co-stimulatory molecules that are required for protection against colitis. PDCs orchestrate beneficial immunoregulatory interaction of commensal microbial molecules with CD4+ T cells through both innate and adaptive immunity.
Macrophage polarization drives granuloma outcome during Mycobacterium tuberculosis infection (IRC5P.475)
Authors: Marino Simeone , Cilfone Nicholas , Mattila Joshua , Flynn JoAnne , Linderman Jennifer , Kirschner Denise 
Published: January 2023

Abstract
Lung granulomas are the pathologic hallmark of tuberculosis (TB), where different immune cell types and bacteria co-localize. Macrophages are the most abundant cells in a granuloma. They are both critical effector and target cells for Mycobacterium tuberculosis (Mtb, the causative agent of TB) infection, and serve as regulators of inflammation and innate immune responses by producing a variety of pro- and anti-inflammatory cytokines. Overall, macrophage function, diversity and location within a granuloma environment are still poorly understood. In this study we used a systems biology approach to better characterize how macrophages are polarized by their microenvironment to mount either a pro- or an anti-inflammatory phenotype. We built on recently published experimental studies on non-human primates that suggest how a continuous dynamic balance between pro and anti-inflammatory signals is achieved to control pathology and restrain bacterial growth within a granuloma. Here we investigated how this balance translates into macrophage polarization and plasticity, and how the relative contribution of these macrophages, broadly classified as ‘M1’ and ‘M2’, evolves over time and leads to an effective immune response, at the granuloma scale. Our in silico granulomas suggest that temporal dynamics of granuloma polarization ratios can be predictive of granuloma outcome. We also find that a signature common to properly formed and functioning granulomas is increased NFkB degradation.
Transferred allogenic S1P3 deficient dendritic cells (DCs) interact with recipient splenocytes to prevent kidney ischemic-reperfusion injury (IRI) (IRC5P.459)
Authors: Bajwa Amandeep , Huang Liping , Ye Hong , Lobo Peter , Okusa Mark 
Published: January 2023

Abstract
The plasticity of DCs permits phenotypic modulation ex-vivo by gene expression or pharmacological agents. Sphingosine-1-phosphate (S1P), a sphingolipid is the natural ligand for S1Pr1-5s. We tested the ability of S1P3-/-(3KO) DC to suppress kidney IRI. BMDCs from B6 WT (B6DC) or 3KOB6DC were adoptively transferred into (-&gt;) Balb/c 1d/7d prior to IRI. Naïve Balb (noDC) had higher plasma creatinine (PCr; mg/dl) compared to sham (1.2±0.17;0.2±0.004,p≤0.01). 3KOB6DCs-&gt;Balb/c were protected from IRI (PCr:0.3±0.07,p≤0.01). 3KOB6DCs-&gt;Balb/c had higher Treg and less CD4 cells compared to naïve or B6DC mice in spleen. The protection afforded by 3KOB6DCs-&gt;Balb/c mice required the recipient mouse to have a spleen; Tregs, CD11c+ DC and T/B cells; Splnx Balb, use of Rag1-/- mice or depletion of recipient Tregs (PC61) or DCs (CD11c-DTR) rendered adoptively transferred 3KODCs ineffective. PKH-26 labeled DCs from WT or 3KO-&gt;Balb/c were located in the marginal zone (MZ) of recipient spleen with close proximity to Tregs. WT and 3KO mice injected with latex beads to demonstrate clearance efficiency showed disruption of MZ in 3KO. Adoptive transfer of 3KOB6DCs prior to IRI attenuates injury through interaction with splenocytes, recipient DCs resulting in induction of Tregs through possible disruption of MZ macs. We conclude that genetically induced deficiency or pharmacological blockade of allogenic S1P3DCs could serve as a useful therapeutic approach to IRI or DGF associated with Transplant.
IL-33 stimulates dendritic cell secretion of IL-2 that promotes selective expansion of ST2+Foxp3+ regulatory T cells (IRC5P.460)
Authors: Matta Benjamin , Lott Jeremy , Mathews Lisa , Rosborough Brian , Blazar Bruce , Turnquist Heth 
Published: January 2023

Abstract
IL-33 is a pleiotropic IL-1 family cytokine that signals via ST2 and expands ST2+Foxp3+ regulatory T cells (Treg) in vivo. As ST2+ Treg show poor expansion by direct IL-33 stimulation, we sought to define mechanisms mediating their expansion. IL-2 signaling promotes ST2 expression on CD4+ T cells, and dendritic cells (DC) express ST2 (able to respond to IL-33), and are a potential source of IL-2. Thus, we examined if IL-33 mediates ST2+ Treg expansion by stimulating DC IL-2 production. CD11c+ wild type (WT) or IL-2 knockout (KO) bone marrow DC were exposed to IL-33 or LPS. DC phenotype was evaluated by multi-color flow cytometric analysis. The influence of IL-33 DC on T cell function was evaluated in MLR with CD4+ T cells, and T cell proliferation and phenotype were determined by flow analysis. Cytokines were quantitated by ELISA. We found that unlike LPS, IL-33 does not influence DC surface phenotype or induce pro-inflammatory cytokine production compared to controls. However, IL-33 induces a 5-fold increase IL-2 production by DC. In MLR, IL-33 DC selectively expand an activated subset of ST2+Foxp3+ Treg that are CD44hiICOShi. IL-33 DC-derived IL-2 is critical since IL-33-exposed IL-2 KO DC fail to expand ST2+ Treg. In summary, IL-33 licenses DC to selectively expand a subset of activated Treg through production of IL-2, in the absence of classical DC activation. These findings may be harnessed to aid the development of novel therapeutics aimed at promoting immune tolerance.
Signaling by ADAP via the Carma1-TAK1 axis is critical for pro-inflammatory cytokine production but not for cytotoxicity in NK cells. (IRM7P.477)
Authors: Rajasekaran Kamalakannan , Kumar Pawan , Schuldt Kristina , Peterson Erik , Vanhaesebroeck Bart , Dixit Vishva , Thakar Monica , Malarkannan Subramaniam 
Published: January 2023

Abstract
‘Cytokine release syndrome’ (CRS) is a serious side effect of NK or T cell-mediated immunotherapy to treat cancer. A strategy to contain CRS without affecting the cytotoxic potential of NK and T cells is highly warranted. A thorough understanding of the signaling pathways will help to identify molecules that exclusively regulate inflammatory cytokine production but not cytotoxicity. Here, we define an ADAP-dependent signaling axis downstream of Fyn that exclusively regulated cytokine production but not cytotoxicity in NK cells. The adapter protein, ADAP is critical for organizing the Carma1-Bcl10-MALT1 signalosome, which is essential for the activation of TAK1. The initiation of these signaling events is critical for the production of Interferon-γ and other pro-inflammatory cytokines. The defect in cytokine production correlated with a decreased nuclear translocation of c-Fos and c-Jun despite a significantly increased phosphorylation of ERK1/2 and JNK1/2 in the ADAP deficient NK cells. Nuclear translocation of NFκB was also decreased in the ADAP deficient NK cells. Our findings indicate that ADAP is indispensible for activating a signaling cascade that leads to inflammatory cytokine production, whereas it is not essential for those signaling events that lead to cytotoxicity. Our findings demonstrate the feasibility of targeting the functions of unique signaling molecules in order to regulate the production of inflammatory cytokines.
Natural killer T cells mediate the protective effects of 1,25-dihydroxyvitamin D3 in experimental autoimmune encephalomyelitis in mice (IRM7P.478)
Authors: Waddell Amanda , Zhao Jun , Cantorna Margherita 
Published: January 2023

Abstract
Active vitamin D (1,25(OH)2D3) completely blocks the development of experimental autoimmune diseases. However, the molecular and immunobiological mechanisms underlying vitamin D’s anti-inflammatory properties are not fully understood. In the present study, we have employed a murine model of experimental autoimmune encephalomyelitis (EAE) in order to determine the role of natural killer T (NKT) cells in vitamin D-mediated protection from EAE. WT mice or mice deficient in all NKT cells (CD1d-/-) or invariant NKT (iNKT) cells (Jα18-/-) were fed control or 1,25(OH)2D3 (50 ng/day) supplemented diets. All mice fed control diet developed severe EAE. 1,25(OH)2D3treatment protected WT mice, but not CD1d-/- or Jα18-/- mice, from developing EAE. Development and severity of EAE were associated with insufficient suppression of the MOG-specific IL-17 response by 1,25(OH)2D3 treatment in CD1d-/- mice compared to WT mice. Activation of iNKT cells with α-galactosylceramide (α-GalCer) or 1,25(OH)2D3 treatment did not protect IL-4-/- mice from developing EAE. Furthermore, 1,25(OH)2D3 treatment of splenocytes in vitro resulted in decreased α-GalCer-induced IL-17 and increased IL-4 production. These studies demonstrate that NKT cells are important mediators of vitamin D-induced protection from EAE in mice and NKT cell-derived IL-4 may be an important factor in providing this protection.
Peroxisome proliferator-activated receptor delta (PPARδ) limits Th1 inflammation through effects on interferon (IFN)-γ transcription and lymphocyte homeostasis (IRM7P.480)
Authors: Zhao Fei , Drohomyrecky Paulina , Dunn Shannon 
Published: January 2023

Abstract
PPARδ is a nuclear receptor that functions as a transcriptional regulator of various processes including metabolic homeostasis, cell proliferation and differentiation. We previously reported that mice deficient in PPARδ (KO) exhibit more severe experimental autoimmune encephalomyelitis than wild type (WT) mice. This more severe inflammation correlated with an increased expansion of myelin-reactive IFNγ-producing cells in the spinal cord, suggesting that PPARδ may act to limit Th1 inflammatory responses. However, the mechanism of how PPARδ accomplishes this is not clear. Characterization of the spleen and thymus compartments revealed that KO mice exhibit reduced spleen and thymus cellularity as compared to WT mice. The resultant lymphopenia in KO mice associated with a higher frequency of memory CD4+ T cells in the periphery. Radiation bone marrow chimeras were generated and the defect in thymocyte development was found to map to the radio-resistant compartment, suggesting that PPARδ may operate in stromal cells to regulate thymocyte development. This was furthered confirmed by the finding that mice with T cell-restricted deficiency of PPARδ did not exhibit defective thymocyte homeostasis. Furthermore, we observed that naïve CD4+ T cell from both KO and T cell-restricted PPARδ-deficient mice had a higher propensity to produce IFNγ upon stimulation. Thus PPARδ appears to limit Th1 inflammation both through the control of lymphocyte homeostasis and IFNγ production.
PTEN regulates natural killer cell trafficking and effector functions (IRM7P.484)
Authors: Leong Jeff , Schneider Stephanie , Sullivan Ryan , Fehniger Todd 
Published: January 2023

Abstract
Phosphatase and tensin homolog (PTEN) is a critical negative regulator of the PI3 kinase pathway, members of which are essential for regulating natural killer (NK) cell signaling and activation. To evaluate whether PTEN was required for normal NK cell functions, we utilized a mouse model of NK specific PTEN deficiency (Ncr1iCre knockin x PTENflox). PTEN deletion resulted in significant loss of NK cells in the bone marrow and other lymphoid tissues, but markedly increased numbers within the peripheral blood and lung. Surprisingly, we observed near equivalent NK cell maturation within peripheral organs regardless of PTEN expression, suggesting that PTEN operates to re-distribute NK cells during development. Similarly, despite impaired migration to chemoattractants in vitro, PTEN-deficient NK cells egress more efficiently from the bone marrow and preferentially reside in sinusoidal compartments, but are specifically retained in the lung and peripheral vasculature. Given the inappropriate localization in the absence of PTEN, we further evaluated whether cytotoxic functions were altered. While PTEN deficient NK cells have enhanced cytotoxicity against co-localized RMAS tumors, they are unable to migrate to distal tumor sites. These data suggest that PTEN is required for the trafficking of NK cells during both homeostasis and in the presence of malignancy, during which it represses cytotoxic functions.
Indoleamine 2,3-dioxygenase inhibits B cell immune response to T independent antigens (IRM8P.710)
Authors: Shinde Rahul , Chaudhary Kapil , Shimoda Michiko , Pacholczyk Gabriela , McGaha Tracy 
Published: January 2023

Abstract
T cell independent (TI) antibody responses are crucial for humoral immunity to viruses and encapsulated bacteria. Here we report a novel mechanism where the tryptophan catabolizing enzyme indoleamine 2,3-dioxygenase (IDO) 1 regulates the B cell response to TI antigens. Immunization with NP-Ficoll led to rapid splenic induction of IDO, primarily in the extra-follicular space. When IDO1KO mice were immunized there was a significant increase in humoral responses with increased formation of extra-follicular IgM and IgG3 foci, antibody secreting cells (ASCs), and increased antibody titers. This was not associated with an alteration in affinity maturation suggesting the primary impact of IDO1 deficiency was increased B cell proliferation and plasma cell formation. IDO1 did not affect immune responses to protein antigens as immunization with NP-OVA elicited similar antibody titers regardless of IDO1 function. In addition, adoptive transfer of IDO1 deficient B cells to µMT-/- (B cell deficient) mice was sufficient to replicate increased TI responses observed in IDOKO mice. Moreover, in vitro LPS rapidly induced IDO1 in MACS-purified B cells and IDO deficient B cells display enhanced LPS and CpG-driven proliferation associated with increased production of IgM, IgG3, and IL10. Thus, our results demonstrate a novel role of IDO in suppressing T cell independent antibody response that provides insight into the understanding of B cell immune responses in autoimmunity and vaccine biology.
Novel ubiquitination-like modifications that regulate recombination of immunoglobulin loci gene segments leading to antibody diversification (IRM8P.711)
Authors: Basu Uttiya , Sun Jianbo , Keim Celia , Grinstein Veronika 
Published: January 2023

Abstract
Programmed DNA mutagenesis events in the immunglobulin (Ig) loci of B cells utilize the common and conserved mechanism of ubiquitination of proteins prior to proteasomal degradation to generate the required antigen-receptor diversity. Recombinase proteins RAG1 and RAG2, required for V(D)J recombination, and activation-induced cytidine deaminase (AID), an essential mutator protein for catalyzing somatic hypermutation (SHM) and class switch recombination (CSR), are regulated by various ubiquitination events affecting protein stability and activity. AID associates with transcribing RNA polII and non-coding RNA processing complex RNA exosome to introduce DNA lesions in the IgH locus that are required for CSR and SHM. Failure in regulating AID’s DNA mutagenic activity is considered as a primary cause for B cell lymphomagenesis, especially Diffuse Large B cell Lymphoma (DLBCL). Here, we present our recent observations that demonstrate that the AID/RNA exosome/RNA polymerase II complex is regulated by ubiquitination and neddylation modifications. Using proteomics based experiments we have identified AID associated protein factors that promote ubiquitin-like modification of AID/RNA polII/RNA exosome complex. Using various biochemical assays, knockdown experiments and genetically mutated mouse models, we demonstrate the ubiquitin-like modification of AID-protein complex is important for regulating AID activity during CSR and overall maintenance of B cell genomic integrity.
Mouse models for immunology research available from the Jackson Laboratory repository. (IRM9P.718)
Authors: Valenzuela Alicia , Lutz Cathleen , Rockwood Stephen , Sasner Michael , Donahue Leah 
Published: January 2023

Abstract
The Jackson Laboratory Repository serves as a centralized facility for the development, rederivation, distribution and cryopreservation of mouse models of human biology and disease. Hundreds of strains are added annually to one of the largest collections of characterized mouse strains available to the biomedical research community. This poster presents new knockout (KO) strains carrying genes associated with T cells and antigen (Ag) presentation. Lcp2 (SLP-76)-null mice lack T cells and γδ-TCR+ cells. Psmb8 (LMP-7) KO mice exhibit impaired Ag presentation and reduced T cells. In Clec9a (DNGR-1) KO/GFP knock-in (KI) mice, cross-priming of CD8+ T cells against dead-cell-associated antigens is altered. Nlrp3-/- (cryopyrin) mice are useful for studying role of the inflammasome in immune responses. New floxed strains include Cxcl12, Csf1r and a floxed-stop ubiquitous Myc-driven human CD2 KI. Cxcl12 functions in lymphoid progenitor and hematopoietic SC maintenance. The cytokine, Csf1r, is associated with macrophage function. Other additions include: a tamoxifen-inducible KI of EGFP/cre/ERT into the Foxp3 locus, a transcription factor required for development and function of Treg cells; a Notch reporter with GFP expression directed to the Notch pathway; an RORgamma-driven cre transgene that directs expression to αβ T cells, immature thymocytes and lymphoid tissue inducer cells. To have your strain considered for inclusion in the Repository please visit www.jax.org/donate-a-mouse.
Cyclic dinucleotides mediate gap junction-dependent spread of DNA-induced IRF3 activation. (IRM9P.729)
Authors: King Kevin , Patel Suraj , Yarmush Martin , Weissleder Ralph 
Published: January 2023

Abstract
Cell death and sterile inflammation are central to the pathogenesis of human diseases such as myocardial infarction and acute liver failure. Since patients often present after injury has begun, a primary therapeutic goal is to limit the extent and duration of injury. Therefore, we investigated the molecular mechanisms by which innate immune responses to dying cells are spread. When a cell dies, it releases danger signals such as DNA, which are sensed by neighboring cells and spread by undefined mechanisms, ultimately defining the extent and duration of injury. To study this process, we created stable monoclonal fluorescent reporter cell lines for key transcriptional regulators of danger signaling (IRF3 and NFkB), exposed them to various danger signals, and monitored their spatiotemporal activation by quantitative fluorescence microscopy. We found IRF3 to be strongly activated not only in cells sensing DNA, but also in their unexposed neighbors. Using pharmacologic inhibitors and genetically modified co-cultures, we found that spread of IRF3 activity is contact-dependent and requires gap junctions. We also provide evidence that the recently described cyclic dinucleotide cGAMP is a gap junction permeable mediator of IRF3 activation spread. In vivo experiments show that genetic or pharmacologic modulation of gap junction communication is protective against acute organ injury, demonstrating the importance of injury spread, and its promise as a therapeutic target.
Endogenous HMGB1 prevents nuclear catastrophe and proinflammatory nucleosomal release (IRM9P.719)
Authors: Tang Daolin , Kang Rui , Lotze Michael , Zeh Herbert 
Published: January 2023

Abstract
The mechanisms by which tissue injury alerts the immune system and drives a systemic inflammatory response syndrome remain unknown. Here we identify a high mobility group box 1 (HMGB1)-mediated nuclear regulatory mechanism for the control of the inflammatory response during tissue injury. We show that conditional knockout of HMGB1 within the pancreas, while benign without provocation, rendered mice dramatically more susceptible to experimental acute severe pancreatitis, as manifested by accelerated tissue injury and animal lethality. Deficiency of endogenous pancreatic HMGB1 led to exaggeration of L-arginine-induced nuclear catastrophe and nucleosome release, which then recruited and activated inflammatory cells with subsequent HMGB1 release locally and into the circulation. Serum levels of tissue enzymes (e.g., amylase, lactate dehydrogenase, and pancreatic myeloperoxidase) and pro-inflammatory cytokines were significantly higher in conditional pancreas-specific HMGB1 knockout mice compared to their wild-type control littermates. Moreover, neutralizing extracellular histone and HMGB1 conferred protection against acute pancreatitis in conditional pancreas-specific HMGB1 knockout mice. Thus, intracellular HMGB1 may serve as a previously underappreciated negative regulator of inflammation, shedding light on the role of the innate immune response in infection and tissue damage.