bluuebunny/crossref_metadata_2025_split huggingface dataset:
SOCS1 regulates IL-4-activated IRS-2 tyrosine phosphorylation in monocytes and macrophages via the proteasome (IRM5P.639)
Authors: McCormick Sarah , Gowda Nagaraj , Carrera Maria , Fang Jessie , Heller Nicola 
Published: January 2023

Abstract
Allergic asthma is a chronic lung disease driven by Th2 cytokines IL-4/-13. In macrophages, IL-4/-13 bind IL-4 receptors, which differentially signal through Insulin Receptor Substrate (IRS)-2, inducing M2 macrophage differentiation. M2 macrophages correlate with disease severity and poor lung function. The mechanisms that regulate M2 polarization are poorly understood. Following IL-4 exposure, Suppressor Of Cytokine Signaling (SOCS)1 is induced in human monocytes. We found that siRNA knockdown of SOCS1 prolonged IRS-2 tyrosine phosphorylation and enhanced M2 differentiation, while knockdown of SOCS3 had no impact. We found human SOCS1 co-immunoprecipitated with IRS-2. Since SOCS1 is an E3 ubiquitin ligase, we examined IL-4-induced ubiquitination of IRS-2. siRNA knockdown of SOCS1 inhibited ubiquitination of IRS-2 while knockdown of SOCS3 did not change IRS-2 ubiquitination. IL-4 stimulation of monocytes from healthy and allergic individuals revealed that SOCS1 is induced in healthy monocytes but not allergic, whereas SOCS3 is induced in allergic monocytes. Healthy monocytes show greater ubiquitination of IRS-2 than allergic monocytes after IL-4 stimulation. Here we identify SOCS1 as critical negative regulator of IRS-2 phosphorylation, limiting IL-4 signaling and M2 differentiation. These findings provide novel insight into the immune bias in allergic monocytes that predispose them to M2 differentiation and present a novel therapeutic target for managing allergic disease.
Long-range chromatin interactions control trophoblast-restricted HLA-G expression during pregnancy (IRM6P.657)
Authors: Ferreira Leonardo , Meissner Torsten , Mikkelsen Tarjei , O'Donnell Charles , Sherwood Richard , Mallard William , Rinn John , Cowan Chad , Strominger Jack 
Published: January 2023

Abstract
Human pregnancy poses an immunological paradox: at the fetal-maternal interface, semi-allogeneic fetal extravillous trophoblasts (EVT) invade the uterine mucosa without being rejected by the maternal immune system. HLA-G is a nonclassical MHC class I molecule specifically expressed on the surface of EVT and is believed to be a key to fetus-induced immune tolerance. However, the EVT-specific expression of HLA-G is still poorly understood. A novel enhancer more than 10 kb upstream of HLA-G, Enhancer L, was discovered via dissection of the HLA-G locus using a Massively Parallel Reporter Assay (MPRA). DNase I Hypersensitive Site (DHS) mapping, Chromosome Conformation Capture (3C) and reporter gene assays established Enhancer L as a cell type-specific enhancer that loops into the HLA-G classical promoter. Strikingly, deletion of Enhancer L using a CRISPR/Cas9 dual guide approach results in complete ablation of HLA-G expression. Saturation mutagenesis of Enhancer L revealed the existence of motifs for transcription factors paramount in trophoblast development. Two of them, GATA2 and CEBPB, associate with Enhancer L and the HLA-G classical promoter, and are absolutely required for HLA-G expression. Interestingly, progesterone upregulates HLA-G, as well as GATA2, via a nonclassical membrane progesterone receptor complex in trophoblasts. These findings shed light on the mechanism by which HLA-G is specifically expressed in invasive trophoblasts during pregnancy.
Reconstitution of TCR/CD3 expression in B cell lymphoma to dissect the contribution of CD3 conformational change from downstream signaling events in the process of TCR/CD3 downregulation. (IRM7P.714)
Authors: Ferrer Alejandro , Reed Brendan , Schrum Adam , Gil Pages Diana 
Published: January 2023

Abstract
T cell receptor (TCR) stimulation induces conformational change in CD3 (CD3Δc), followed by phosphorylation of ITAMs that leads to receptor downregulation among other T cell responses. To begin to dissect the contribution of CD3Δc from downstream signal transduction mechanisms associated with these events, we have transduced the mouse B cell lymphoma X63.Ag8.653, which lacks expression of the src-family kinase Lck required for proper ITAM phosphorylation, with the OT-I TCR chains Vα2 and Vβ5, together with all CD3 subunits and the coreceptor CD8αβ, generating the cell line X63.OT1.CD8. Stimulation of the OT-I TCR with anti-CD3 antibody or the cognate antigen SIINFEKL-H2Kb induces CD3Δc in this cell line. As expected by the absence of Lck, CD3 ITAM phosphorylation was undetectable in our cell line and no calcium mobilization was found upon antibody stimulation. Unexpectedly, stimulation with either SIINFEKL-H2Kb or anti-CD3 did induce TCR/CD3 downregulation in our cells. These data suggest CD3ε PRS exposure via CD3Δc might be sufficient to transduce some signals that contribute to TCR downregulation in the absence of CD3 ITAM phosphorylation.
A systematic investigation on the mechanism of non-catalytic tyrosine phosphorylated receptor triggering (IRM7P.718)
Authors: Denham Eleanor , Goyette Jesse , van der Merwe P. 
Published: January 2023

Abstract
Non-catalytic tyrosine phosphorylated receptors (NTRs) are a large class of leukocyte receptors that bind to surface-associated ligands and include both activating and inhibitory members. They contain, or associate with adaptor molecules which contain, tyrosine residues within conserved cytoplasmic motifs that are phosphorylated and dephosphorylated by extrinsic kinases and phosphatases respectively. The mechanism by which NTR-ligand engagement leads to sustained phosphorylation of receptor tyrosine-based motifs and initiation of downstream signalling (‘receptor triggering’), is as yet unknown. Our hypothesis is that the NTRs signal using the kinetic segregation (KS) model. To systematically test whether predictions of the KS model hold for all the NTRs we have developed an artificial generic ligand system in which biophysical and biochemical properties of NTR-ligand interactions can be manipulated, including: NTR-ligand dimensions, ligand densities and NTR-ligand affinities. This system uses a Strep-tag II attached to the ectodomain of NTRs. We demonstrate that representative NTRs, both activating and inhibitory, can be triggered through ligation of this tag. In agreement with a key KS model prediction we further show that only ligation by surface-associated ligands leads to triggering, whilst ligation by soluble ligands does not. The flexibility of this system will allow us to further test other predictions of the model, in addition to other parameters of NTR signalling.
Identifying the precursor of natural IgM-producing plasma cells (LYM2P.736)
Authors: Reynolds Alexander , Kuraoka Masayuki , Kelsoe Garnett 
Published: January 2023

Abstract
Natural antibody is constitutively present in the serum, and plays an important role in the clearance of cellular debris, the early control of viral and bacterial expansions, and the initiation of T-dependent humoral responses. However, despite widespread recognition of the critical role of natural IgM in the prevention of both infectious and autoimmune diseases, the source of natural antibody long remained obscure. We have found that most serum IgM in resting mice is produced by a discrete population of IgM plasma cells in the bone marrow, a substantial fraction of which are long-lived cells that occupy a survival niche distinct from that required by IgG plasma cells. Natural IgM plasma cells originate from precursors in the peritoneal cavity, as transfer of unfractionated peritoneal washout cells completely restores serum IgM and the specific compartment of bone marrow plasma cells in Rag1-deficient mice. Surprisingly, however, transfer of purified peritoneal B1a, B1b, or B2 cells did not efficiently reconstitute serum IgM, while a distinct peritoneal B-cell population reconstituted both natural IgM and peritoneal B1a cells in Rag1-deficient mice. We propose that natural IgM plasma cells represent an “innate B-cell” population descended from yolk sac progenitors, and that natural IgM plasma cells are not the progeny of B1a cells per se but instead share an earlier common ancestor also present in the peritoneal cavity.
NKAP is required for invariant natural killer T cell development and generation of ROR-γt expressing iNKT17 (LYM2P.738)
Authors: Thapa Puspa , Chen Meibo , McWilliams Doug , Belmonte Paul , Constans Megan , Hiebert Scott , Sant'Angelo Derek , Shapiro Virginia 
Published: January 2023

Abstract
iNKT cells differentiate into functional subsets defined as NKT1, NKT2 and NKT17, characterized by their expression of transcription factors, T-bet, GATA3 and ROR-γt and production of IFN-γ, IL-4 and IL-17, respectively. In our earlier studies, we showed that the transcriptional repressor NKAP is required for positive selection of DP thymocytes into the iNKT lineage, using CD4-cre NKAP conditional knockout (cKO) mice. To study the role of NKAP in subsequent stages of iNKT cell development and differentiation, we generated PLZF-cre NKAP cKO mice. In these mice, NKAP deletion occurs after entry into the iNKT lineage at stage 0, bypassing the requirement for NKAP in iNKT development at the DP stage. PLZF-cre NKAP cKO mice have a significant reduction in the absolute number of iNKT cells starting at Stage 1. This block in development is not due to defect in cell homeostasis or increased cell death, but is due to decreased rate of proliferation. PLZF-cre NKAP cKO mice have very few T-bet expressing NKT1 cells with similar amount of IFN-γ production to WT, whereas there was almost no ROR-γt expressing NKT17 cells with no IL-17 production. There is a decrease in the absolute number of NKT2 cells; however, IL-4 production by NKAP-deficient NKT2 cells is similar to WT. Thus, NKAP is required for proliferation of iNKT cells early in development, and is required for the differentiation of iNKT cells into the NKT17 lineage.
Reduced T follicular helper function in aged mice is correlated with increased numbers of T regulatory cells (LYM2P.725)
Authors: Masters April , Lefebvre Julie , Qiu Zhijuan , Hopkins Jake , Haynes Laura 
Published: January 2023

Abstract
T follicular helper (TFH) cells are critical for a germinal center response and a robust humoral response. Aging has a profound impact on the formation of a strong humoral/germinal center response, yet the impact of aging on TFH remains unclear. This study explores the effects of aging on TFH transcription factors, which could explain some of the age related germinal center defects. During influenza infection, young mice exhibit a very robust IgG response, which is significantly reduced in aged mice. Initial studies showed that there are no differences in the number of flu-specific TFH (CXCR5hi PD-1hi) during the course of the infection, as determined using MHC class II NP tetramer. To determine transcription factor expression and localization of TFH, we performed multicolor flow cytometry and confocal microscopy on spleens from these mice. There was no difference in BCL6 expression in young and aged flu-specific TFH. But, strikingly, flu-specific BCL6hi TFH from aged mice had increased co-expression of Foxp3, characteristic of T follicular regulatory cells, a critical controller of germinal center responses. We are currently exploring the contribution of whether the differences in TFH transcription factor expression are intrinsic to aged CD4+ T cells or are a result of the aged microenvironment. These results strongly suggest that aging has a dramatic impact on transcription factor expression in CD4 T cells, which can influence the establishment of a robust immune response.
Reduction of regulatory T cells during acute Toxoplasma gondii infection is caused by IFN-γ (MPF2P.759)
Authors: Olguin Jonadab , Fernández Jacquelina , Salinas Nohemí , Terrazas Luis , Saavedra Rafael 
Published: January 2023

Abstract
Toxoplasmosis is an infection disease caused by the protozoan parasite Toxoplasma gondii. A TH1 immune response is essential for protection against the parasite, IFN-γ being the major mediator of resistance. CD4+Foxp3+ Regulatory T cells (Treg) maintain immune homeostasis by actively suppressing pathological and physiological immune responses. Intraperitoneal infection of BALB/c mice with T. gondii cysts leads to high mortality rate and exacerbated IFN-γ levels in sera both of which parallel a decreased Treg number in blood and spleen. Since we observed an inverse correlation between high IFN-γ levels and low Treg cell number in infected mice, we analyzed if this cytokine was involved in Treg reduction. Since STAT-1 is involved during signaling by IFN-γ, we infected STAT-1-/- KO mice with T. gondii and found that the Treg cell number was unchanged in blood and increased in spleen, although all mice died; IFN-γ levels in sera in these mice were lower than those observed in infected WT mice, and CD4+ - and CD8+-producing IFN-γ cells were also reduced. These results strongly suggested that IFN-γ has a major role in the reduction of Treg cells. Thus we injected a neutralizing mAb against IFN-γ to infected Foxp3EGFP transgenic mice and unchanged Treg cell number was detected in blood. Our results thus demonstrate that exacerbated IFN-γ levels during acute toxoplasmosis are responsible of collapse of Treg cell number in this model.
Tumour necrosis factor-related apoptosis inducing ligand promotes the development of experimental chronic obstructive pulmonary disease (MUC1P.905)
Authors: Hansbro Philip , Haw Tatt , Nair Prema , Hanish Irwan , Nguyen Duc , Liu Gang , Inman Mark , Kim Richard , Collison Adam , Knight Darryl , Yagita Hideo , Mattes Joerg , Horvat Jay , Starkey Malcolm 
Published: January 2023

Abstract
Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide and causes significant healthcare and economic burden. Cigarette smoking is a major risk factor. There is a lack of effective treatments for COPD due to the poor understanding of the underlining mechanisms. Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is implicated in respiratory diseases such as asthma and pulmonary fibrosis. However, the role of TRAIL in the pathogenesis of COPD is unknown. In this study, TRAIL mRNA expression and/or protein levels in the lung (airway and parenchyma) and serum were increased in a mouse model of cigarette smoke-induced experimental COPD. Genetic deletion of TRAIL significantly reduced cigarette smoke-induced pulmonary inflammation, expression of key pro-inflammatory mediators, emphysema-like alveolar enlargement and improved lung function in experimental COPD. Interestingly, genetic deletion of TRAIL led to spontaneous small airway remodelling characterized by increased airway epithelial cell thickness and collagen deposition. Importantly, antibody-mediated neutralization of TRAIL reduced cigarette smoke-induced pulmonary inflammation, emphysema-like alveolar enlargement and small airway remodelling. Our study is the first to show that TRAIL plays an important role in the pathogenesis of COPD and provides further evidence for TRAIL being a pivotal inflammatory cytokine in respiratory diseases.
Effect of anti-high mobility group box 1 on innate lymphoid cells of dry eyes in mouse model of Sjögren syndrome (MUC2P.927)
Authors: Kim Mee , Jeong HyunJeong , Jin Suk Ryu , Kim Dong Hyun , Wee Won Ryang 
Published: January 2023

Abstract
How innate lymphoid cells (ILCs) secrete cytokines is not well understood. High mobility group box (HMGB) may be involved in secreting IL17 or IL22 in ILCs. Extracellular HMGB1 may be related with the pathogenesis of Sjögren syndrome. We here investigated that administration of anti-HMGB1 might attenuate dry eyes by modulating ILCs in mouse model of Sjögren syndrome. This study was conducted in conformance with the FASEB Statement of Principles for the use of animals in research and education. 10 weeks-old NOD. B10 or B6 mice had been subconjunctivally injected by 0.02 to 2 ug of anti-HMGB1 antibodies or PBS for 2 weeks. Injection of anti-HMGB1 attenuated corneal epithelial erosions significantly by fluorescein staining and increased tear secretion transiently by phenol-red thread test. After stimulation of the cells from Peyer’s patches or cervical drainage lymph nodes by IL1b and IL23, cells were negatively gated using anti-mouse CD3-PerCP-Cy5.5 and B220-FITC, and were positively gated using anti-mouse CD45-APC-Cy7, NKp46-APC and IL22-PE. The percentages and the numbers of NKp46+IL22 or NKp46-IL22 secreting cells were increased in cervical lymph nodes of anti-HMGB1 injected mice compared with those in PBS injected mice. While, those were not significantly changed in Peyer’s patches of anti-HMGB1 or PBS injected mice. Together, data demonstrate that administration of anti-HMGB1 attenuates dry eyes by increasing IL-22-secreting ILCs in mouse model of Sjögren syndrome.
Modeling and implications of antibody transcytosis by human genital epithelial cells during Chlamydia trachomatis infection and challenge (MUC2P.930)
Authors: Albritton Hannah , Buckner Lyndsey , Taylor Stephanie , Lillis Rebecca , Martin David , Kozlowski Pamela , Quayle Alison 
Published: January 2023

Abstract
Chlamydia trachomatis (CT) is a common sexually transmitted pathogen; there is no vaccine and infection can lead to infertility and increased susceptibility to HIV acquisition. An obligate intracellular bacterium, CT is tropic for genital columnar epithelial cells, and the endocervix is the primary site of infection in women. CT infections are usually asymptomatic, endure for months, and immunity is short-lived. However, recent epidemiology studies have identified a subgroup of women who spontaneously clear infection without treatment and are protected from re-infection. We hypothesize that genital secretions from these women will aid in the identification of site-specific immune correlates and inform vaccine design. Since animal and clinical studies suggest antibodies may play a role in protection and clearance of CT, we developed novel clinical sampling techniques and an ELISA that allows normalization and quantification of local CT-specific antibody isotypes from CT-infected women, and report significantly higher levels of CT-specific IgG/A in genital secretions compared to serum. Development of a novel in vitro endocervical epithelial model that can be infected with CT now allows us to (i) determine the protective efficacy of defined (clearer v non-clearer) patient samples against CT-challenge and (ii) explore recent observations that inflammation modulates endocervical immunoglobulin receptor expression, likely influencing antibody transport in the CT-infected cervix.
Intestinal epithelial cell mediated miscommunication in type 1 diabetes: a tipping point for disease? (MUC2P.933)
Authors: Graves Christina , Lapato Melissa , Wallet Shannon 
Published: January 2023

Abstract
The host-environment dialogue at the intestinal interface has been strongly implicated in the development of inflammatory diseases. We are interested in understanding the role of this dialogue in autoinflammatory and autoimmune diseases such as type 1 diabetes (T1D). Although the gastrointestinal tract, a dynamic and highly regulated immunologic organ, has been demonstrated to be impaired in T1D, the mechanisms of any (mis)communications taking place remain unknown. Here we describe the global intestinal inflammatory environment and the phenotype of intestinal lymphocytes in human T1D. In addition, we evaluated the intestinal epithelial cell (IEC) innate immune responses using primary cell culture. Our data indicate a whole organ reduction in tolerogenic mediators concomitant with elevated IEC-induced pro-inflammatory mediators. Significant alterations in intestinal leukocyte populations were observed, whereby increased frequencies of pathogenic T cells and alterations in tolerogenic DCs were noted. Finally, T1D-derived primary IEC cultures demonstrated a lack of tolerogenic responses concomitant with induction of inflammatory response to microbial ligand stimulation as compared to non-T1D derived cultures. Together these data suggest a loss of gastrointestinal homeostasis in T1D potentially as a result of a dysfunctional IEC-mediated host-environment dialogue, though whether this is a cause of consequence of disease remains to be defined.
Effect of 17-allylaminogeldanamycin on antimicrobial peptide and cytokine expression in HaCaT cells after IL-17 and heat stimuli (MUC2P.941)
Authors: Kim Bo-Kyung , Park Minhwa , Kim Ji-Yon , Lee Kyung Ho , WOO SOYOUN 
Published: January 2023

Abstract
The pathogenesis of inflammatory skin disease involves interactions between Th17-mediated immune response and keratinocytes. Immersion in pools of thermal mineral water is often considered to have chemical, thermal, mechanical, and immune modulatory benefits to inflammatory skin disease. Therefore, we questioned the effect of thermal treatment on IL-17-mediated immune response in keratinocytes. We evaluated the effect of HSP90 inhibitor on IL-17-mediated cytokine and antimicrobial peptide expression in keratinocytes after heat treatment including IL-1β. To examine the effect of IL-17 receptor-mediated expression of antimicrobial peptides or inflammatory chemokines in HaCaT cells, Act1 siRNA-transfected HaCaT cells were heat stimulated at 42oC and 1 uM of 17-allylaminogeldanamycin (17-AAG) was added. At 37oC, Act1 knockdown decreased BD-1, LL-37, S100A8, S100A9, CXCL1, CXCL5, and CXCL8 expression; however, expression of these antimicrobial peptides and chemokines increased after incubation at 42oC. This thermal-mediated effect was suppressed upon treatment with 17-AAG. We also found that IL-1β expression decreased by 17-AAG after heat stimulation. Although HSP90 enhanced the expression of antimicrobial peptides or chemokines in keratinocytes after heat treatment, HSP90 inhibitors reversed these effects. Therefore, HSP90 inhibitors after heat stimulation (e.g., thermal baths) may be a beneficial therapeutic approach for IL-17-mediated inflammatory skin disease.
Candida albicans morphology and DC subsets determines T helper differentiation (MUC2P.920)
Authors: Igyarto Botond , Kashem Sakeen , Gerami-Nejad Maryam , Kumamoto Yosuke , Mohammed Javed , Jarrett Elizabeth , Drummond Rebecca , Zurawski Sandra , Zurawski Gerard , Berman Judith , Iwasaki Akiko , Brown Gordon , Kaplan Daniel 
Published: January 2023

Abstract
Candida albicans is a dimorphic fungus responsible for chronic mucocutaneous and often fatal systemic infections. Mucocutaneous immunity to C. albicans requires Th17 differentiation that is thought to depend on recognition of filamentous C. albicans while systemic resistance is considered T cell independent. Using a murine skin infection model, we compared in vivo T helper responses to yeast and filamentous C. albicans. Unexpectedly, we found that only yeast induced Th17 responses through a mechanism that required Dectin-1 mediated expression of IL-6 by Langerhans cells. Filamentous forms induced Th1 without Th17 due to the absence of Dectin-1 ligation. Notably, Th17 responses provided selective protection against secondary cutaneous infection while Th1 provided protection against systemic infection. Thus, C. albicans morphology drives distinct Th responses that provide tissue specific protection. These findings provide novel insight into compartmentalization of Th responses, C. albicans pathogenesis and have critical implications for vaccine strategies.