bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of inflammasome cytokines IL-1 and IL-18 during infection with neurotropic mouse hepatitis virus (INC8P.434)
Authors: Zalinger Zachary , Weiss Susan 
Published: January 2023

Abstract
The inflammasome is a cytosolic multiprotein complex that mediates secretion of pro-inflammatory cytokines IL-1 and IL-18. These cytokines have been found to play a role in restriction of a wide range of pathogens, and to contribute to the pathogenesis of many autoimmune conditions. IL-18 signaling has a known role in promoting production of interferon gamma from T and natural killer cells, while IL-1 polarizes Th17 responses and recruits neutrophils to sites of infection. However, work with influenza A and West Nile virus have shown that IL-1 signaling can also be necessary to promote strong anti-viral T cell responses via activation of dendritic cells. This effect was found to be specific to the site of infection. To further elucidate the roles of these cytokines we have infected mice lacking the IL-1 or IL-18 receptor with a dual neuro- and hepatotropic strain of mouse hepatitis virus (MHV). IL-18 receptor deficient mice succumb to normally non-lethal doses of MHV while IL-1 receptor deficient animals are somewhat protected from lethal infection, suggesting the cytokines play opposite roles to each other. The T cell response is similar in all genotypes, suggesting the cytokines act on innate populations of cells. We are currently investigating the behavior of the neutrophil and natural killer cell populations in inflammasome cytokine deficient mice. This work should elucidate the effect of inflammasome signaling on innate immune responses in differing sites of infections.
Diminished IL-12 production mediates impaired T-cell stimulating capacity of lymph node dendritic cells and macrophages in SIV infection (INC9P.444)
Authors: Wonderlich Elizabeth , Barratt-Boyes Simon 
Published: January 2023

Abstract
Mononuclear phagocytes are integral components of the innate immune system that sense invading microbes and expand antigen-specific immune responses. Thus far, their roles in progressive HIV infection remain to be clarified. We used the pathogenic simian immunodeficiency virus (SIV) rhesus macaque model to determine the immune stimulating capacity of plasmacytoid dendritic cells (pDC), myeloid DC (mDC), and macrophages (Mφ) after SIV infection, focusing on the lymph node compartment. By positive selection through live-cell sorting, we measured the ex vivo ability of each subset to respond to uridine-rich ssRNA oligos derived from the infecting strain of SIV and to subsequently drive CD4+ T cell proliferation and IFN-γ production. Isolated pDC and Mφ from infected macaques were defective at stimulating CD4+ T cells to divide and produce IFN-γ. Co-stimulatory marker upregulation on mononuclear phagocytes remained comparable in naïve and infected samples, whereas deficiencies in IFN-α and IL-12 production by pDC and Mφ were found concurrent with lower CD4+ T cell function. Furthermore, exogenous IFN-α and IL-12 supplementation was able to recover the CD4+ T cell stimulating capacity of isolated mDC and Mφ from SIV-infected lymphatic tissue. These novel findings suggest that defects in mononuclear phagocyte T cell stimulating capacity through loss of IL-12 production may contribute to the overall immune dysfunction in pathogenic SIV of rhesus macaques and HIV infection of humans.
Protein kinase D1 links IRAK1 and TRAF6 in the MyD88-dependent Toll-like receptor signaling pathway. (INM9P.457)
Authors: Yi Ae-Kyung , Park Jeoung-Eun , Kim Young-In 
Published: January 2023

Abstract
The initial proinflammatory responses to Saccharopolyspora rectivirgula (SR), which causes farmers' lung disease, is dependent on Toll-like receptor 2 (TLR2) and MyD88, and MyD88 is absolutely required for SR-induced protein kinase D1 (PKD1) activation. We found that SR-mediated proinflammatory responses and neutrophil infiltration in the lungs are ablated in mice treated with PKD inhibitor or lacking the PKD1 gene. However, the mechanisms by which PKD1 is activated and functions to regulate TLR signaling are incompletely understood. In the present study, we found that upon TLR ligand stimulation, the receptor signaling complex composed of MyD88, IRAK4, IRAK1, TRAF6, and PKD1 is formed in membrane lipid rafts and that IRAK4 and IRAK1, but not TRAF6, are required for recruitment of PKD1 to the receptor complex and for activation by TLR ligands. In addition, MyD88-dependent translocation of TRAF6 into lipid rafts, ubiquitination of TRAF6, degradation of IRAK1, and activation of TAK1 are dependent on PKD1. Our findings indicate that PKD1 is a downstream effector of IRAK1, but an upstream modulator of TRAF6. Together with the known ability of PKD1 to shuttle between membrane lipid rafts and the cytosol, our results further support the hypothesis that PKD1 functions as a molecular shuttle that brings TRAF6 to IRAK1 in membrane lipid rafts and then aids in translocation of the IRAK1/TRAF6 complex to the cytosol, where TRAF6 interacts with its downstream effectors.
Innate immune-induced neuronal cell apoptosis via a SARM1 mediated mechanism in the absence of viral replication. (INM9P.451)
Authors: Taylor Katherine , Woods Tyson , Peterson Karin 
Published: January 2023

Abstract
Neuronal cell death is a hallmark of multiple neurological diseases including viral infections. The innate immune response in the CNS can contribute to neuronal cell death indirectly through the production of cytokines and neurotoxins as well as via the recruitment of immune cells. However, the direct induction of neuronal cell death via innate immune mechanisms is less well characterized and often confused by confounding factors such as viral replication. We previously identified sterile alpha and HEAT/Armadillo motif containing 1 (SARM1) as a mediator of bunyavirus induced neuronal apoptosis. Currently, we have identified a new mechanism of SARM1-mediated neuronal cell death independent of viral infection of the cell. Treatment with TLR7 and TLR9 agonists induces significant levels of neuronal cell death in cortical neurons. Cell death was associated with induction of SARM1 and inhibition of SARM1 suppressed TLR7 and TLR9 mediated neuronal apoptosis. SARM1 localized to the mitochondria and induced oxidative damage leading to neuronal apoptosis. Mechanistic studies indicated that SARM1 co-localized with both PINK1 and UXT proteins indicating a putative mechanism of SARM1 mediated neuronal cell. Thus, activation of pattern-recognition receptors on neurons in the absence of active virus infection can lead to neuronal death and the mechanism of death is via production of SARM leading to mitochondrial damage.
Mechanisms underlying specificity of natural killer cell regulation of adaptive immunity (IRC8P.485)
Authors: Waggoner Stephen , Hatfield Steven , Brooks Taylor , Karmele Erik , Peppa Dimitra , Rydyznski Carolyn , Daniels Keith , Welsh Raymond 
Published: January 2023

Abstract
Viral persistence is determined in part by the regulatory activities of murine natural killer (NK) cells, including the lysis of activated CD4 T cells. We now find that NK cells also impair virus-specific memory T cell responses [and repress humoral immunity (separate abstract)] after acute Pichinde or lymphocytic choriomeningitis virus infections. Hence, limiting NK cell inhibition of adaptive immunity during vaccination could enhance long-lived protective immunity. To enable selective targeting of this NK cell activity in a vaccine, the molecular mechanisms of such suppression must be ascertained. Thus, we have explored changes in the expression of NK cell receptors (NKR) and their ligands (NKRL) on both NK cells and target T cells after infection. This analysis revealed that: 1) suppression of adaptive immunity corresponded with reduced inhibitory NKR expression on NK cells; 2) archetypical activating NKR such as NKG2D were dispensable in our system; and 3) susceptibility of activated CD4 T cells to NK cell lysis correlated with reduced expression of inhibitory NKRL (e.g. CD48) and corresponding higher levels of activating NKRL (e.g. Ly108), than were present on NK cell-resistant CD8 T cells. Further, the selectivity of NK cell killing of different T cells could be deregulated by altering NKR/NKRL expression. Our findings may facilitate development of innovative strategies to harness or inhibit NK cell lysis of T cells in the context of vaccination or immune therapy.
Consumption of plant-derived agonists that enhance NK cell activity is beneficial in infection- and DSS- induced colitis (IRC8P.486)
Authors: Snyder Deann , Hedges Jodi , Robison Amanda , Kimmel Emily , Jutila Mark 
Published: January 2023

Abstract
Inflammatory bowel disease (IBD) is characterized by chronic, relapsing inflammation for which there is no known cure and limited treatment and preventative options. Plant products in many nutritional supplements have been shown by multiple groups to be protective in IBD. We have identified polysaccharides from Acai berries (Acai PS) and polyphenols from apple peels (APP) that specifically stimulate γδ T cells, as well as NK cell and monocyte populations. The effects are similar in some respects to responses induced by patterns derived from the microbiota. Mice fed Acai PS, but not APP, had increased IL-12 expression in spleens, MLN and serum compared to water fed mice and displayed no adverse inflammatory effects in response to the fed supplements. Ingestion of both APP and Acai PS protected from dextran sulfate sodium (DSS)-induced colitis. Acai PS-, but not APP- fed mice with colitis had increased expression of NKG2A, an NK cell receptor implicated in protection from DSS-induced colitis. Acai PS also protected against Salmonella Typhimurium-induced enterocolitis. Novel oral innate agonists Acai PS and APP protect against colitis, but perhaps through distinct mechanisms. Augmented immune signaling is likely to contribute to maintenance of the epithelial barrier and underlying immunity. Considering the paucity of therapeutic and prophylactic options, these plant derived agonists may represent novel therapeutic options for patients with IBD.
2B4(CD244) is required for optimal activation of iNKT cells through CD48 ligation on iNKT cells. (IRC8P.489)
Authors: Shin Jung Hoon , Park Se-ho 
Published: January 2023

Abstract
iNKT cells are a specialized subset of T cells which can modulate immune responses in a diverse way and require fine regulation of their own reactivity. 2B4 (CD244) is a member of the SLAM family molecules and acts as both inhibitory and activating receptor in murine CD8+ T and NK cells depending on relative expression levels of CD48 on target cells, whereas human 2B4 is known to be an activating molecule. Although iNKT cells also express 2B4, the function of 2B4 on iNKT cells remains largely unknown. Here, we report that the optimal activation of iNKT cells requires 2B4. iNKT cells in 2B4 knock-out (KO) mice showed impaired activity and the symptoms of Con-A induced hepatitis were substantially weaker in 2B4 KO mice than those in wild type mice. 2B4 on iNKT cells also required CD48 engagement to exert its effect. Strikingly however, we found that CD48 providers were not necessarily target cells. These results suggest that 2B4 on iNKT cells is required for the optimal stimulation of iNKT cells and 2B4/CD48 interaction of iNKT cells can be mediated through homotypic cell-cell interaction
Peptidase inhibitor 16 identifies a unique subset of memory T helper cells with hyperproliferative and proinflammatory properties (IRC8P.477)
Authors: Mohandas Arunesh , Zola Heddy , Barry Simon , Krumbiegel Doreen 
Published: January 2023

Abstract
T helper (Th) cells play a major role in protecting the body against pathogens. Any imbalance in Th cell subsets could lead to autoimmune and inflammatory diseases. Peptidase inhibitor 16 (PI16), also known as prostate secretory protein of 94 amino acid - binding protein, was recently discovered to be expressed on memory regulatory T cells. This study investigates the expression and function of PI16 on Th cells. In healthy adults, 5-25% of CD4+ Th cells express PI16 with over 90% showing a memory phenotype. PI16+ Th cells have an increased expression of chemokine receptors CCR4 and CCR6 compared with PI16- Th cells. Transwell migration assays showed that more PI16+ Th cells migrated towards the CCR4 and CCR6 ligands (CCL17 and CCL20) compared with PI16- Th cells. After 7 day stimulation using CD3 / CD28 beads, PI16+ Th cells produce more IL-17A and less IFN-g compared with PI16- Th cells. PI16+ Th cells also have a higher expression of ROR-gt compared to PI16- Th cells. Furthermore, in comparison to PI16- Th cells, PI16+ Th cells have an increased proliferative potential and are less responsive to suppression by Treg. The memory phenotype of PI16+ Th cells, the high expression of Th17-like chemokine receptors, increased ROR-gt expression and high production of IL-17A suggest an active role of PI16+ Th cells at the site of infection or inflammation. Further studies are ongoing to understand the functional role of PI16 on Th cells in autoimmune and inflammatory diseases.
Two distinct subsets of skin-infiltrating memory T cells, TRM and TEM, define cutaneous immune responses against systemically-administered soluble antigens (IRC9P.702)
Authors: Egawa Gyohei , Miyachi Yoshiki , Kabashima Kenji 
Published: January 2023

Abstract
Cutaneous immune reactions to soluble antigens, such as drugs, are common, but their precise pathomechanism remains unclarified. To this end, we newly developed a murine model for skin eruptions against soluble antigens using ovalbumin (OVA)-specific TCR transgenic line, OT1. OT1 T cells were adoptively transferred to naïve mice and were immunized with OVA/CFA. Five days later, a skin inflammation was induced to the right ear with topical PMA treatment, which induced skin-resident memory T cells (TRM). Two weeks after PMA-painting, a large number of CD103+CD69+ OT1-TRM were found in the epidermis of PMA-painted site, whereas only a marginal number of OT1 T cells were detected in the untreated left ear. Then, OVA peptide was injected intravenously, which exhibited immediate ear swelling response only in the right ear where OT1-TRM exist in the epidermis. Five days after the systemic administration of OVA peptide, CD44+CD62L- effector memory OT1 T cells (TEM) were significantly increased in the blood and, intriguingly, were detected in the dermis of the untreated left ear. In addition, a repeated intravenous administration of OVA peptide developed chronic necrotic skin lesion even without extrinsic stimuli. These observations implicate respective roles of TRM and TEM in cutaneous immune responses: antigen-specific TRM in the epidermis cause immediate and localized skin eruptions, whereas TEM infiltrate from the blood into the dermis and induce chronic disseminated dermatoses.
Utilizing dual-costimulation through CD134 and CD137 with varying levels of antigen presentation to elicit heterogeneous CD4+ T cell responses (IRC9P.703)
Authors: Morales Crystal , Maxwell Joseph , Adler Adam , Vella Anthony 
Published: January 2023

Abstract
Many cancer vaccines target CD8 T cells, but cytotoxic CD4+ T cells are emerging as important players. Our lab utilized a dual-costimulation immunotherapy approach through CD134 (OX40) and CD127 (4-1BB) agonist antibodies together with T cells specific to a widespread antigen which induced cytotoxic CD4+ T cell differentiation. Drawing upon a novel transgenic mouse model where CD11c+ cells present a specific antigen, we now demonstrate that antigen-specific dual-costimulated CD4+ T cells can be heterogeneous with differential expression of Eomes, KLRG1, GzmB, and the IL-33R (ST2). Specifically, in one transgenic mouse line with lower levels of CD11c antigen presentation, CD4+ T cells developed into Eomes+KLRG1+GzmB- cells. Despite comparable costimulation however, CD4+ T cells developed into Eomes-KLRG1-GzmB+ cells in a mouse line with greater levels of CD11c antigen presentation. Analysis of known helper T cell transcription factors and cytokines also showed expression of other helper T cell subset markers in these cells. Further, the capacity of these cells to become activated and kill target cells is currently being investigated with an in vitro assay. These findings are novel, demonstrating that cytotoxic CD4+ T cells may in fact be a heterogeneous population influenced by antigen presentation levels. Thus, this data should greatly impact immunotherapeutic targeting of cytotoxic CD4+ T cells.
Human thymopoiesis and naïve T cell maintenance throughout life (LYM7P.724)
Authors: Thome Joseph , Ventevogel Melissa , Ohmura Yoshiaki , Kato Tomoaki , Sempowski Gregory , Farber Donna 
Published: January 2023

Abstract
The extent to which thymopoieis is diminished over the human lifespan and how this affects the naïve T cell repertoire in peripheral tissue sites is not well characterized. In collaboration with the New York Organ Donor Network, we have attained novel access to thymus and secondary tissues from deceased organ donors. From this, it is possible to correlate naïve T cell development to distribution in tissue sites from the same individual effectively identifying when functional thymopoiesis is halted and how this affects the naïve T cell pool over time. We have found marked cessation of functional thymopoiesis in donors over 40 years of age but strikingly naïve T cells are maintained in lymph node tissue sites for decades undergoing low levels of proliferation and turnover comprising over 20% of total T cells in donors aged 40-73. When profiling double negative (DN) stage thymocytes using CD25 and CD44, DN1 thymocytes accumulate in donors with negligible thymic output indicating a lack of progenitor cells to progress to maturation while identified recent thymic emigrants (RTE) exhibit differential distribution and regulation in tissues during periods of active thympopoiesis but are evenly distributed in the spleen and lymph nodes when the export of new naïve T cells is halted. Together this indicates that naïve T cell populations are uniquely maintained as a function of thymic activity in order to preserve the ability to respond to new pathogen throughout the human lifetime.
Lck is essential in the non-canonical T cell receptor CR signaling pathway regulated pressed by Cbl during T cell development (LYM7P.727)
Authors: Chiang Yungping , Hodes Richard 
Published: January 2023

Abstract
A canonical pre-TCR/TCR signaling pathway has been extensively characterized that involves sequential phosphorylation and signaling through Lck, ZAP-70, LAT, and SLP-76, and that plays a critical role in thymic T cell development as well as TCR-mediated activation of mature T cells. However, recent studies have reported that genomic deletion of Cbl substantially reverses the defect in thymic development that is seen in ZAP-70-deficient mice. To understand this ZAP-70 independent non-canonical pre-TCR/TCR signaling pathway, we have characterized the effect of Cbl inactivation on the physiologic and signaling defects in mice deficient in the proximal signaling molecules Lck and ZAP-70. We demonstrated that inactivation of Cbl reversed the defects in phosphorylation of multiple signaling molecules, including ERK, PLC-γ1 and AKT, in TCR-stimulated ZAP-70-/- thymocytes, and partially reversed the defective phenotype of T cell development in ZAP-70-/- mice. In contrast, inactivation of Cbl was unable to reverse either defective phosphorylation of TCR signaling molecules or the defective T cell developmental phenotype of Lck-/- mice. Lck thus plays a critical role in both the canonical TCR signaling pathway and in a ZAP-70-independent non-canonical pathway that is normally inhibited by Cbl but is capable of mediating productive thymocyte development.
iNKT cell development is modulated by TCR avidity (LYM7P.728)
Authors: Mallevaey Thierry 
Published: January 2023

Abstract
Invariant Natural Killer T (iNKT) cells are versatile innate T cells that influence immune responses associated with infection, cancer, inflammation and autoimmunity, notably through the production of a wide array of cytokines and chemokines. iNKT cells respond to lipid antigens presented by the MHC Class Ib molecule CD1d. iNKT T cell receptors (TCRs) have limited diversity, due to exclusive usage of a canonical Vα14-Jα18 TCRα chain and a limited set of TCR Vβ gene segments, but with extensive CDR3β diversity. Recent structural and mutational studies revealed that CD1d-lipid-TCR interactions differ markedly from classical pMHC-TCR interactions. Remarkably, docking onto CD1d-lipid complexes is unaffected by the nature of the lipid involved, nor TCRβ composition (Vβ gene segment usage or CDR3β diversity). Rather, we and others have demonstrated that TCRβ composition modulates TCR avidity for CD1d-lipids. Here, we have generated TCR retrogenic mice using TCRs with various avidities, and found a strong correlation between the frequency and numbers of iNKT cells that develop in vivo and TCR avidity for CD1d-lipids. In addition, TCR strength impacts on iNKT cell maturation as well as cytokine production upon immunization with the prototypical lipid αGalCer. In summary, our results highlight a crucial role for iNKT TCR strength on the selection and functional programing of iNKT cells.
Immune perturbation in patients with TGFbeta pathway defects (LYM7P.729)
Authors: Tran Dat , Regalado Ellen , Milewicz Dianna 
Published: January 2023

Abstract
Intro: Mutations in TGFbR1, TGFbR2 and SMAD3 are associated with familial thoracic aortic aneurysms and aortic dissections (TAAD). These patients offer an opportunity to study their immune development. Methods: PBMC from TAAD (n=9) and controls (CT, n=8) were analyzed by FACS. Th1 (IFNg) and Th17 (IL17A) were determined with intracellular cytokine staining. Foxp3+ Tregs were detected with anti-Foxp3. CD19+ were analyzed for naive (IgD+CD27-), unswitched (IgD+CD27+) and switched memory (IgD-CD27+). Plasmacytoid (CD303+pDC) and myeloid (CD1c+mDC) were defined within lineage-1 negative population. Results: %CD3-CD16+NK, CD3+CD16+NKT, CD4+, CD8+ and CD4+CD45RA+ in TAAD were similar to HC. Average %CD19+ (20.8vs7.3, p=0.006) and naive B cells (81.3vs66.6, p=0.004) were higher in TAADvsCT. The unswitched were similar but the switched B cells were lower (8.6vs15.5, p=0.01). While the %Tregs were similar, there was a remarkable reduction (1/2-3 folds, p&lt;0.05) in Foxp3 concentration based on median fluorescence intensity of Foxp3 in TAAD. There was a significant reduction of %Th17 (0.14vs0.61, p=0.01), while the Th1 were similar. %pDC (9.4vs24.1, p=0.009) and %mDC (11.4vs17.9, p=0.01) were also lower in TAAD. Conclusions: These results demonstrate the involvement of TGFbeta signaling in B cells, DCs, Th17 and Treg development. Further studies and monitoring of the clinical effects of these immunological perturbations are needed to appreciate the impact of their underlying disease.