bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inflammation and decreased immunoregulation characterize the leptin receptor deficient (db/db) mouse model of type II diabetes. (173.12)
Authors: Hardie Virginia , Wheeler Matthew , Harms Robert , Boyer Craig , Thiessen Kevin , George Nick , Targy Natalie , Lorenzo Kristina , Mercer David , Sarvetnick Nora 
Published: January 2023

Abstract
Type II diabetes is an inflammatory disease marked by chronic immune activation. Our goal is to describe the fundamental distinctions in the immune system of the db/db mouse. Serum was collected for a 58-factor immunoassay. Splenic leukocytes were isolated for six panels of flow cytometry to examine immune subsets and activation states. Db/db mice exhibit higher levels of proinflammatory factors such as C-reactive protein, granulocyte chemotactic protein-2, monocyte chemotactic protein-1, macrophage inflammatory protein-2, IL1α, and T-RANTES and lower levels in regulatory IL10. Flow cytometry data revealed upregulation of the adhesion molecule CD11b on neutrophils, monocytes and macrophages. Macrophages also expressed increased levels of costimulatory molecule CD80. Inflammatory DC were increased while regulatory DC were decreased. Pre-germinal center B cells were decreased. Plasma and memory B cells were increased with an increase in CD22 maturity marker. FoxP3+ T regs were decreased with a reciprocal increase in CD127. Total CD8 T cells were increased with greater naive populations. Unactivated NK cells were increased, and all NK populations had decreased Ly49H and KLRG1. Above results were significant with p&lt;0.05. Baseline immune activation in db/db mice is marked by increased circulating proinflammatory factors, decreased circulating regulatory factors, increased activation of innate and humoral immunity, and suppressed regulatory and cellular immunity.
The role of sphingosine-1-phosphate receptor-1 in antigen-specific CD8 T cell migration (173.14)
Authors: Bénéchet Alexandre , Maher Leigh , Hla Timothy , Khanna Kamal 
Published: January 2023

Abstract
Our previous studies have helped illuminate the landscape in which the endogenous CD8 T cell immune response occurs. However, the factors that regulate the anatomical program that antigen-specific CD8 T follow after infection are not well understood. Although, it is known that sphingosine-1-phosphate receptor 1 (S1P1) controls naïve T cell trafficking, our understanding of how S1P1 regulates the migration and differentiation of CD8 T cells after infection remains poor. Here we investigated the role of S1P1 in the migration and localization of antigen-specific CD8 T cells in the lymph node (LN) after viral infection. We utilized pharmacological modulation of S1P1 and cell-specific S1P1 knock out mouse models to reveal a central role of S1P1 in controlling effector CD8 T cell migration from the draining LN. In the absence of S1P1 receptor signaling, antigen-specific CD8 T cells failed to utilize the lymphatic sinuses to egress from the draining LN. Furthermore, we used a novel mouse model that contains a mutated native S1P1 protein that cannot be efficiently internalized. Using this mouse model we show that receptor internalization can regulate effector CD8 T cell response early after infection. Therefore, our studies demonstrate that S1P1 receptor signaling is the dominant factor that regulates endogenous effector CD8 T cell migration in secondary lymphoid organs.
Dendritic cells act through IL-6 to enhance naïve T cell trafficking across high endothelial venules during adaptive immunity (173.18)
Authors: Fisher Daniel , Muhitch Jason , Vardam Trupti , Evans Sharon 
Published: January 2023

Abstract
Dendritic cells (DC) are as master regulators of the adaptive immune response, providing both cognate antigen and costimulation required for T cell activation as well as cytokines (e.g., IL-12, TNF, and IL-6) that drive T cell expansion and differentiation. While the role of DCs in T cell activation is well known, here we show that DCs play an unexpected role in shaping the T cell repertoire within inflamed lymph nodes (LN) by promoting trafficking across vascular gateways termed high endothelial venules (HEV). Immunization of mice with bone marrow-derived DCs matured ex vivo by TLR4 and 9 agonists (LPS and ODN-CpG, respectively), but not a TLR2 agonist (Pam3CSK4), increased the intravascular density of two hallmark adhesion molecules, peripheral lymph node addressin (PNAd) and intercellular adhesion molecule-1 (ICAM-1), selectively in HEVs of inflamed LNs. Increased PNAd and ICAM-1 display occurred rapidly after immunization (within 12 h) and coincided with improved trafficking of naïve and central memory T cells as well as increased LN size. PNAd and ICAM-1 expression reverted to homeostatic levels 24-48 h post immunization. Remarkably, increased adhesion molecule expression was abrogated by loss of a single cytokine, IL-6, from immunizing DCs. These findings suggest a model whereby IL-6-producing DC actively promote trafficking in HEVs, thereby increasing the probability that naïve T cells come into contact with cognate antigen-loaded DCs in inflamed LN.
Cognate antigen is necessary and sufficient for the migration of effector and memory T cells to vascularized allografts (173.19)
Authors: Walch Jeffrey , Zeng Qiang , Hoffman Rosemary , Oberbarnscheidt Martin , Camirand Geoffrey , Lakkis Fadi 
Published: January 2023

Abstract
A key step in rejection is the transendothelial migration (TEM) of Ag-specific effector and memory (E&M) T cells into the graft. It is assumed that this step is mediated by Gαi-coupled chemokine receptor signaling. This migratory paradigm was revisited by treating Ag(OVA)-specific(OT-I) and non-specific(P14) CD8 E&M T cells with pertussis toxin (PTx) to block Gαi signaling or not and co-transferring to B6 recipients of B6 Act-OVA or wt vascularized allografts to investigate the roles of cognate Ag and Gαi signaling in T cell TEM. Flow cytometry quantification demonstrated that OVA-specific(OT-I) E&M T cells migrated only to heart grafts that expressed cognate Ag(OVA) and their migration was not inhibited by PTx. Non-specific(P14) T cells, on the other hand, did not migrate to OVA grafts unless OT-I T cells were present and their migration was abrogated by PTx. Two-photon intravital imaging of transplanted OVA kidneys established that PTx only inhibited the firm adhesion and TEM of non-specific(P14) E&M T cells and not the Ag(OVA)-specific(OT-I) E&M T cells. The migration of Ag-specific E&M T cells to vascularized allografts requires cognate Ag and is independent of chemokine signaling. Entry of non-specific E&M T cells, on the other hand, requires chemokines but occurs only if Ag-specific T cells are present. These findings underscore a critical role for cognate Ag in directing the entry of primed T cells to non-lymphoid tissues and suggest novel mechanisms for T cell TEM.
Acute cigarette smoke exposure modulates the inflammatory response in pdmH1N1 and H9N2 infection (P6190)
Authors: Han Yan , Ling Manto , Mao Huawei , Tu Wenwei , Lau Yulung 
Published: January 2023

Abstract
Although the health risks of cigarette smoke are widely reported, the impact and mechanism of short-term cigarette smoke exposure on inflammatory diseases are still unclear. Until now, there is no study about the role of cigarette smoke in 2009 pandemic H1N1 and avian influenza A H9N2 infection. Based on our previous study, which investigated the mouse mortality, weight changes, lung virus loads and inflammatory cytokines and chemokines’ expression, we further analyzed the status of inflammatory responses before the virus infection, pulmonary histological analysis and the lung cell profiles after virus infection. Mild inflammatory responses were shown before influenza A virus infection after cigarette smoke exposure. For pdmH1N1 infection, although there was a trend that smoke exposure group had a lower lung inflammatory score and less neutrophils infiltrated into the lung than the air exposure group, the difference was not significant. However, after H9N2 virus infection, the mice in smoke exposure group displayed a significantly milder disease, as evidenced by a lower lung inflammatory score, as well as less macrophages, neutrophils, CD4+ T cells and CD8+ T cells infiltrated into the lung compared to the air exposure group. Our study provided the first in vivo evidence that acute cigarette smoke exposure may modulate the effect of influenza A virus infection. The underlying mechanism is under investigation.
The N-terminal region of the guanine nucleotide exchange factor Vav1 plays a distinguished role in T cell receptor mediated calcium signaling (P1158)
Authors: Cao Youjia , Li Shiyang , Du Ming-Juan , Wan Ya-Juan , Lan Bei , Liu Yao-Hui , Yang Yin , Zhang Cui-zhu 
Published: January 2023

Abstract
Vav1 is a guanine nucleotide exchange factor specifically expressed in hematopoietic cells. It consists of multiple structural domains and plays important roles in T cell activation. The other highly conserved isoforms of Vav family, Vav2 and Vav3, are ubiquitously expressed in human tissues including lymphocytes. All the three Vav proteins activate Rho family small GTPases, which are involved in a variety of biological processes during T cell activation. Intensive studies have demonstrated that Vav1 is indispensable for T cell receptor mediated signal transduction. T cells lacking Vav1 exhibited severe defect in TCR-mediated calcium elevation, indicating that the co-existing Vav2 and Vav3 did not compensate Vav1 in calcium signaling. What is the functional particularity of Vav1 in lymphocytes? In this study, we identified the N-terminal region of Vav1 in calponin homology domain to be essential for its interaction with Calmodulin, that leads to TCR-induced calcium mobilization. Substitution of the region of Vav1 with the homologous region of Vav2 or Vav3 abolished the association with CaM, and the N-terminal mutations of Vav1 failed to potentiate normal TCR-induced calcium mobilization, that in turn, suspended nuclear factor of activated T cells activation and IL-2 production. This study highlights the importance of the N-terminal region of Vav1 for calmodulin binding, and that attributes irreplaceable role of Vav1 in T cell activation and signal transduction.
Ca2+ regulates T cell receptor activation by neutralizing the charge of phospholipids (P1164)
Authors: Xu Chenqi 
Published: January 2023

Abstract
The inner-leaflet of the plasma membrane is enriched of anionic phospholipids, which interact with many positively charged proteins and regulate their function. The cytoplasmic domains of CD3ϵ and ζ chains in TCR-CD3 complex are enriched of basic residues and they interact with the inner-leaflet of the plasma membrane in quiescent T cells. Crucial tyrosine residues in ITAM insert into the hydrophobic core of the membrane, preventing the spontaneous tyrosine phosphorylation by Lck. An obvious follow-up question is what would dislodge ITAM from the membrane after TCR engagement. Ligation of TCR with agonist peptide-MHC can induce receptor clustering, which can disrupt ITAM-membrane binding and trigger the initial TCR phosphorylation and downstream signaling events including Ca2+ influx. As a divalent cation, Ca2+ directly binds to the phosphate group of phospholipids, which can disrupt the ionic interaction between CD3ϵ/ζ cytoplasmic domains and the plasma membrane and facilitate ITAM phosphorylation. This regulation can help the phosphorylation of bystander TCRs and amplify the weak initial ligand-induced TCR signaling. When compared with wild-type T cells, T cells deficient of CRAC channel have substantially lower TCR phosphorylation level after triggering. Our study thus demonstrates a positive feedback regulation of Ca2+ to TCR phosphorylation involving lipid manipulation, which helps explain the unique nature of T cell’s hypersensitivity to even a single antigen molecule.
Transcription factor Ascl2 limits Tfh cell differentiation and function (IRC8P.493)
Authors: Bertino Sarah , Dong Xuemei , Laidlaw Brian , Craft Joe 
Published: January 2023

Abstract
Follicular helper T (Tfh) cells promote germinal center (GC) formation and subsequent B cell maturation through the secretion of cytokines such as IL-21 and IL-4. Achaete-scute complex homologue 2 (Ascl2), a transcription factor previously shown to have a role in stem cell maintenance, is highly upregulated in the Tfh-cell subset of CD4+ T cells. However, its role in Tfh-cell development and function has not been established. We generated mice with conditional deletion of Ascl2 in the CD4 T cell compartment to assess whether this protein has a functional role in Tfh cells. Tfh cells deficient in Ascl2 promote a more robust germinal center response, with larger germinal centers and an increased percentage of GC B cells after immunization. This phenotype was coupled with augmented class-switched antibody production in the Ascl2-deficient mice. The expansion of GCs and class-switched antibody production is likely a result of increased IL-4 production by Tfh cells in the Ascl2-deficient mice. These data suggest that Ascl2 may play a role in limiting the functionality of Tfh cells by preventing their secretion of IL-4 and repressing their potential as cell helpers.
In vivo monitoring of the immune response to mIgG conjugated to iron oxide nanoparticles using the pulp (dermis) of growing feathers as a test-tissue in chickens, (IRC8P.495)
Authors: Alaamri Olfat , Byrne Kristen , Falcon Daniel , Erf Gisela 
Published: January 2023

Abstract
Biomedical applications of nanoparticles (NPs) are growing. Data on the biological activities of NPs are, however, limited. We are evaluating the growing feather (GF) in chickens as a test-site (US-Patent 8,216,551; GF Erf) to monitor in vivo activities of NPs. In this study, the immune response to mouse (m) IgG initiated with different antigen-adjuvant combinations was examined. Specifically, 8-week-old male chickens were immunized i.m. with 0.1 mL of mIgG alone, a mixture of Alum adjuvant & mIgG, or mIgG conjugated to iron oxide NP (IONP-mIgG, 10 nm) (n=6 birds/treatment). To monitor the immune response, 0.01 mL mIgG was injected into the pulp (dermis) of 20 GF/bird ten days post-immunization. GF were collected before (0 h) and at 0.25, 1, 2, 3, 5, and 7 d post-GF-injection for flow cytometry, mRNA and histological analysis. Flow-cytometric cell-population analysis of pulp cell suspensions revealed higher levels (%, P&lt;0.001) of lymphocyte infiltration [due to B cells (P&lt;0.001), alpha-beta TCR+ T cells (P&lt;0.005) and CD4+ T cells (P&lt;0.05)] in birds immunized with IONP-mIgG compared to Alum & mIgG or mIgG. These data demonstrate the effectiveness of GF as a test-site to monitor qualitative and quantitative aspects of immune activities in vivo and the usefulness of this minimally invasive procedure as a window into biological activities of NP. Moreover, the results suggest effective adjuvant activities of IONP in stimulating a primary immune response to protein antigen.
Tim-3 enhances CD8 T cell responses to acute listeria monocytogenes infection (IRC9P.711)
Authors: Gorman Jacob , Starbeck-Miller Gabriel , Pham Nhat-Long , Rothman Paul , Traver Geri , Harty John , Colgan John 
Published: January 2023

Abstract
Tim-3 is a cell surface molecule expressed on multiple immune cell populations that can mediate both stimulatory and inhibitory effects. Tim-3 has been previously shown to enforce CD8 T cell exhaustion, a dysfunctional state associated with chronic stimulation. In contrast, the role of Tim-3 in the regulation of CD8 T cell responses to acute infection remains undefined. To address this knowledge gap, we examined how Tim-3 affects CD8 T cell responses to acute Listeria monocytogenes (LM) infection. Analysis of wild-type (WT) mice infected with LM showed that Tim-3 was transiently expressed by activated CD8 T cells and was associated with acquisition of an effector CD8 T cell phenotype. Comparison of responses to LM by WT and Tim-3 KO mice showed that the absence of Tim-3 significantly reduced the magnitudes of both primary and secondary CD8 T cell responses, which correlated with decreased IFN-γ production and degranulation by Tim-3 KO cells. To address the T cell-intrinsic role of Tim-3, we analyzed responses to LM infection by WT and Tim-3 KO TCR-transgenic CD8 T cells following adoptive transfer into a shared WT host. In this setting, the generation of cytokine-producing cells and proliferation by Tim-3 KO cells were significantly reduced, demonstrating that Tim-3 has a direct effect on CD8 T cell function. Our results suggest that Tim-3 can mediate a stimulatory effect on CD8 T cell responses during an acute infection.
T Cell Factor-1 and β-catenin specifically and critically control the generation and function of T helper 2-type natural killer T cells (LYM7P.725)
Authors: Berga Bolanos Rosa , Sharma Archna , Sultana Dil , Chen Qinghua , Heller Nicola , Sen Jyoti 
Published: January 2023

Abstract
Phenotypically and functionally heterogeneous natural killer T (NKT) cells develop from common thymocyte precursors. Transcriptional programs that regulate the development of different subsets of NKT cells remain to be delineated. In this report we demonstrate that transcription factors T cell factor (TCF)-1 and β-catenin direct the development of CD4+ T helper (Th)2-type, IL-4-producing NKT2 cells in the thymus without influencing the development of Th1-type NKT1 cells. NKT2 cells are significantly diminished in TCF1-deficient and β-catenin-deficient mice and are increased in mice with enforced expression of β-catenin. Increased NKT2 cells in mice with enforced expression of β-catenin are functional and produce increased cytokines that exacerbate the development of IL-25-dependent airway hypersensitivity.
FoxP3+Helios+, but not FoxP3+Helios-, Tregs express GARP and LAP, representing the expanded subpopulation in pancreatic cancer patients (TUM2P.911)
Authors: Elkord Eyad , Al Samid Abd Al Samid , Khaled Yazan , Ammori Basil 
Published: January 2023

Abstract
Recent evidences show that Tregs negatively impact anti-tumour immunity and cancer immunotherapy. We, and others, reported that Tregs are expanded in peripheral blood and tumour microenvironment of cancer patients, correlating with poor prognosis and reduced survival. Our work has further established that Treg infiltration of tumors is correlated with a lack of patients’ responsiveness to therapy. Recent observations indicate that Tregs could be phenotypically and functionally different in cancer settings. However, molecules contributing to their enhanced immunosuppressive activity are yet to be elucidated. We recently found that the majority of peripheral and tumor-infiltrating FoxP3+ Tregs expresses Helios, an Ikaros family transcription factor. Interestingly, FoxP3+Helios+ Treg subset is significantly higher in cancer patients, compared to healthy individuals. Herein we show that markers of activated Tregs [latency associated peptide (LAP) and glycoprotein A repetitions predominant (GARP, or LRRC32)] are mainly expressed on FoxP3+Helios+ Treg subset, but not on FoxP3+Helios- Treg subset. Additionally, FoxP3+Helios+GARP+LAP+ activated Tregs represent the expanded subset in pancreatic cancer patients, compared to controls. Taken together, these observations confirm that the expanded FoxP3+Helios+ Tregs in cancer patients represent the activated Treg subset, with more suppressive characteristics, that could be targeted by specific protocols.
CTLA-4+ regulatory T cells are increased in cetuximab treated head and neck cancer patients, suppress NK cell cytotoxicity and correlate with poor prognosis (TUM2P.916)
Authors: Jie Hyun-Bae , Schuler Patrick , Lee Steve , Srivastava Raghvendra , Argiris Athanassios , Ferrone Soldano , Whiteside Theresa , Ferris Robert 
Published: January 2023

Abstract
The EGFR-targeted antibody, cetuximab, is clinically effective against head and neck cancer (HNC) but only in 15 - 20% of patients. We hypothesized that regulatory T cells may have functional and clinical impact antitumor immunity induced in cetuximab-treated patients. Indeed, Treg frequency and expression of CTLA-4, CD39, and surface TGF-β were significantly increased after cetuximab-based therapy. These Treg were inversely associated with functional activity of intratumoral NK cells and with clinical outcome in cetuximab-treated HNC patients. Importantly, NK cells selectively eliminated intratumoral Treg in the presence of anti-CTLA mAb (ipilimumab), which overcame the Treg suppressive activity on cetuximab-mediated ADCC. These results suggest that differences in Treg-mediated suppression may play a role in the clinical response to cetuximab therapy, and that efficacy of cetuximab therapy could be enhanced by adding ipilimumab to eliminate Treg, thus promoting anti-tumor immunity.
Tissue resident memory T cells are found in the primary and metastatic tumors of breast cancer patients (TUM2P.922)
Authors: Egelston Colt , Lee Peter 
Published: January 2023

Abstract
Tumor tissues are composed of significant numbers of Tumor Infiltrating Lymphocytes (TILs), which may include CD8+ T cells, CD4+ T cells, and B cells. Of the T cell population, it is known that these cells are often functionally suppressed and defective in their ability to kill and proliferate. However, little is known about how T cells traffic into tumor sites. Furthermore, it is unclear how the various T cell memory subtypes play a role in attempting to eradicate tumor cells. Such knowledge is critical for the development of successful cancer immunotherapies. Here we assayed peripheral blood mononuclear cells (PBMCs), tumor tissue, and sentinel lymph nodes (SLNs) of breast cancer patients to phenotype memory T cells. We found higher proportions of EM cells among CD8 TILs as compared to SLN CD8 T cells. Conversely, the CD4 compartment in both TILs and SLNs was dominated by CM CD4 T cells. Further analysis showed that breast cancer patient CD8+ TILs in primary tumors were made up of a significant population of Tissue Resident Memory cells (TRMs) as identified by CD103 expression and high CD69 expression. We also showed that the CD8+ TILs found in a metastatic brain lesion of a breast cancer patient also contained TRMs. Finally we showed that healthy breast tissue from a patient with no clinical signs of breast cancer also contained CD8+ TRMs.
CpG immunization protects against a lethal influenza challenge and generates antigen specific memory T cells (VAC2P.932)
Authors: Vogel Alexander , Condon Shirley , Brown Deborah 
Published: January 2023

Abstract
Despite extensive research, influenza A virus (IAV) remains a major cause of mortality and healthcare expenditure. Emerging pandemics from highly pathogenic IAV strains such as H5N1 and H1N1 highlight the need for universal, cross protective IAV vaccines. Current vaccine formulations generate strain-specific neutralizing antibodies primarily against the outer coat proteins hemagglutinin and neuraminidase. In contrast to these highly mutable proteins, internal proteins of IAV are more conserved and are therefore a favorable target for developing vaccines that induce strong T cell responses in the absence of humoral immunity. Here, we found that intranasal administration of a TLR 9 agonist (CpG) and inactivated x31 (H3N2) protects mice from a heterosubtypic challenge with a lethal dose of PR8 virus (H1N1). Vaccinated mice also showed less weight loss and lower viral titers compared to control mice following a lethal PR8 challenge. Both CD4 and CD8 effector functions were measured five days post challenge with significant increases in the number of Granzyme B positive cells as well as antigen specific IFN-γ cells. Additionally, vaccinated mice had higher memory recall responses following ex vivo restimulation with NP specific peptides as measured by IL-2 and IFN-γ production. Thus, CpG is a potent inducer of antiviral immunity highlighted by robust T cell responses underscoring the potential of this vaccination strategy as a prophylactic measure against pandemic IAV strains.