bluuebunny/crossref_metadata_2025_split huggingface dataset:
Granzyme B contributes to impaired wound healing during chronic inflammation in apolipoprotein E knockout mice (P3036)
Authors: Hiebert Paul , Wu Dan , Granville David 
Published: January 2023

Abstract
Granzyme B (GzmB) is a serine protease expressed by immune cells during chronic inflammation and is capable of degrading extracellular matrix (ECM) proteins such as fibronectin (FN). Apolipoprotein E (ApoE) is a protein known to suppress inflammation and is expressed in skin. ApoE knockout (KO) mice develop an inflammatory skin phenotype when fed a high fat diet leading to impaired wound healing. We hypothesized that excessive GzmB activity contributes to impaired healing in ApoE KO mice through the degradation of ECM. Wild type (WT), ApoE KO and ApoE/GzmB double knockout (DKO) mice were fed a high fat diet for 30 weeks and given a 1 cm diameter skin wound on their mid dorsum. Wounds were allowed to heal for 16 days at which point the wounded tissue was harvested. All WT mouse wounds showed complete closure after 16 days while only 40% achieved closure in ApoE KO mice. Wound contraction was also slower in ApoE KO mice compared to WT controls (P&lt;0.05). Histological analysis demonstrated persistent inflammation in ApoE KO mouse wounds at Day 16 featuring neutrophils, macrophages, T cells and GzmB. Wounded skin from ApoE KO mice also contained a FN fragment of similar size to that produced by GzmB in vitro. Compared to ApoE KO mice, DKO mice showed faster contraction (P&lt;0.05) and better healing with 80% of wounds achieving closure by Day 16. These results suggest GzmB contributes to impaired wound healing in ApoE KO mouse wounds, possibly through excessive degradation of FN.
TSLP elicits IL-33-independent innate lymphoid cell responses to promote skin inflammation (P3040)
Authors: Kim Brian , Siracusa Mark , Saenz Steven , Noti Mario , Monticelli Laurel , Sonnenberg Gregory , Hepworth Matthew , Van Voorhees Abby , Comeau Michael , Artis David 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) are a recently identified family of immune cells that regulate inflammation and tissue repair at barrier surfaces such as the intestine and lung. Previous studies have demonstrated that group 2 ILCs found in the intestine or lung respond to the predominantly epithelial cell-derived cytokines IL-33 and IL-25 and produce the type 2 cytokines IL-5 and IL-13. However, whether group 2 ILCs are found in the skin, whether they respond to epithelial cell-derived cytokines and whether they contribute to skin inflammation has not been characterized. Here, we identified a population of skin-resident group 2 ILCs present in healthy human skin and enriched in lesional human skin isolated from atopic dermatitis (AD) patients. Group 2 ILCs were also found in normal murine skin, accumulated in the skin and skin-draining lymph nodes in a murine model of AD-like inflammation and were critical for the elicitation of type 2 cytokine responses and AD-like inflammation. Remarkably, ILC responses and AD-like inflammation were independent of IL-33 and IL-25 but critically dependent on thymic stromal lymphopoietin (TSLP). Collectively, these results demonstrate an essential role for IL-33- and IL-25-independent group 2 ILCs in promoting skin inflammation.
An 8-color panel for detection of human blood dendritic cells by flow cytometry (P3014)
Authors: Miloud Tewfik , Dupas Nathalie , Montero-Julian Felix 
Published: January 2023

Abstract
Dendritic cells (DCs) are antigen presenting cells capable of presenting antigen and priming a T cell response. They form a heterogeneous group of cells based on phenotype, location and function. In human blood, DCs represent less than 1% of white blood cells, and can be separated into 2 main cell subsets, namely the myeloid DCs (mDCs) and the plasmacytoid DCs (pDCs). Among the mDCs, 3 distinct cell subsets are identified: CD1c+mDCs, CD141+mDCs and CD16+mDCs. In blood, the frequency of DCs is affected in certain pathological conditions such as HIV, diabetes, asthma, chronic viral hepatitis, and graft-versus-host-disease. Thus, the detection and enumeration of different blood DC subsets is important to understand immune regulation in pathological conditions and to guide specific patient treatments. Due to the lack of specific markers for DC definition, the combination of several markers is required to allow their identification. Based on current knowledge in human DC biology, we have evaluated the expression and association of several DC markers to design an optimized 8-color panel for flow cytometry which allows for the detection of all DCs subsets in whole blood samples or peripheral blood mononuclear cells (PBMCs). This panel (CD1c/HLADR/Lineage/CD11c/CD16/Clec9A/CD123/CD45) provides an easy and robust assay to study the role of DCs in healthy donors and patient samples.
Fli-1 transcription factor regulates inflammatory mediators in endothelial cells (P6240)
Authors: Karam Eva , Williams Sarah , Zhang John 
Published: January 2023

Abstract
Fli-1 transcription factor is implicated in the pathogenesis of human and murine lupus. A two-fold overexpression of Fli-1 protein in transgenic mice results in the development of a lupus-like disease. Expression of Fli-1 in lupus patients is higher compared to normal controls. Endothelial cells play a role in a number of biological processes ranging from inflammation to the regulation of blood flow. We have reported that Fli-1 directly regulates the expression of MCP-1 in endothelial cells. In this study we investigated if Fli-1 also regulates other inflammatory chemokines. We transfected an endothelial cell line with Fli-1 specific siRNA and negative control oligo siRNA. There was more than an 80% decrease in Fli-1 expression in cells transfected with Fli-1 siRNA when compared to those transfected with control siRNA. Next, we stimulated these cells with TLR4 ligand lipopolysaccharide and extracted mRNA. An mRNA array was used to screen the levels of 26 chemokines. A number of message levels were decreased, including MCP-1, in the cells transfected with Fli-1 specific siRNA when compared to the control. Expression of MIF was higher in the cells when Fli-1 was silenced. We further confirmed MIF mRNA expression levels by real-time PCR. Our data indicates that Fli-1 is likely involved in regulating the production of several chemokines in endothelial cells.
Isoliquiritigenin and glycyrrhizin inhibit the inflammasome activation in a different manner. (P1246)
Authors: Honda Hiroe , Nagai Yoshinori , Takatsu Kiyoshi 
Published: January 2023

Abstract
A soponin glycyrrhizin (GL) and a chalcone isoliquiritigenin(ILG) are representative components of Glycyrrhiza uralensis, which attenuate inflammatory responses mediated by TLR4. We have showed that GL and ILG suppressed different steps of the LPS sensor TLR4/MD-2 complex signaling at the receptor level. GL and ILG attenuated LPS binding to the TLR4/MD-2 complex and LPS-induced TLR4 homodimerization, respectively. In the present study, we examined effects of ILG and GL on IL-1β production in macrophage. When bone marrow-derived macrophages were stimulated with LPS to prime TLR4 signaling pathway followed by ATP to activate NLRP3 inflammasome, significant IL-1β production was observed. However, addition of ILG or GL together with ATP severely impaired the IL-1β production without changing the pro-IL-1β expression levels. We infer from these results that ILG and GL inhibit the NLRP3 inflammasome activation in both priming phase as well as activating phase. We also investigated effects of ILG and GL on the AIM2 inflammasome activation and found that GL but not ILG could suppress its activation, while both ILG and GL could suppress ATP-dependent inflammasome activation.We will discuss molecular mechanisms by which these compounds suppress inflammasome activation.
NFkB1 inhibits NOD2-induced cytokine secretion through ATF3-dependent and -independent mechanisms (P1252)
Authors: Zheng Shasha , Abraham Clara 
Published: January 2023

Abstract
NOD2, an intracellular sensor of bacterial peptidoglycan, is associated to Crohn’s disease (CD). In the intestine, NOD2 is continuously stimulated. Upon chronic NOD2 stimulation in human monocyte-derived macrophages (MDM), cytokine secretion is significantly attenuated upon restimulation of NOD2 or other pattern recognition receptors (PRR); cytokine downregulation is a prominent feature of intestinal tolerance. NFkB1 (p50/p105) is a critical transcription factor regulating PRR responses. However, the role of NFkB1 in NOD2-mediated tolerance reported is not known. Knocking down NFkB1 expression in primary human MDM significantly reversed NOD2-induced cytokine downregulation. MAPK and IkBa activation was dramatically reduced in NOD2-initiated tolerance; this reduced signaling was reversed upon NFkB1 knockdown. Furthermore, upon NOD2-initiated tolerance, NFkB1 knockdown significantly reversed the regulation of both activating (H3K4Me2, H4Ac) and inhibitory (H3K27Me3) histone modifications within cytokine gene promoters. Upon chronic NOD2 stimulation, the transcriptional repressor, ATF3, was induced and bound to the promoters of cytokine genes; both features were reversed upon NFkB1 knockdown. Restoring ATF3 expression under these NFkB1 knockdown conditions significantly restored NOD2-initiated tolerance. We therefore define a clear role for NFkB1 in the mechanisms through which the CD-associated protein NOD2 downregulates cytokines.
IL-12 priming and selective IL-2Rα engagement leads to enhanced CD8+ T cell persistence and anti-tumor immunity in the absence of lymphodepletion. (P1424)
Authors: Rubinstein Mark , Moore Caitlin , Su Ee , Songalia Neizel , Neitzke Daniel , Cloud Colleen , Johnson C. , Paulos Chrystal , Cole David 
Published: January 2023

Abstract
Effective T cell therapy against metastatic cancer requires high frequencies of functional tumor-reactive T cells. However, achieving sustained and elevated levels of tumor-reactive T cells has proven difficult and usually requires highly toxic preconditioning of patients with radiation or chemotherapy to destroy host lymphocytes and suppressive cells. Using the mouse CD8+ T cell pmel-1 TCR transgenic model, we found that the combination of 1) in vitro IL-12-priming of donor T cells during activation and 2) the administration of IL-2/anti-IL-2 mAb complexes after adoptive transfer promoted the dramatic and systemic increase in tumor-reactive T cells in the absence of host lymphodepletion. Importantly, this combinatorial therapy led to the cure of over half of mice with established subcutaneous B16 tumors. Interestingly, while IL-2/anti-IL-2 mAb complexes synergized with ex vivo IL-12-priming, IL-15/sIL-15Rα-Fc complexes failed to mediate comparable synergistic activity despite signaling through the shared IL-2Rβγ subunits. Given that IL-12 priming strongly promotes the upregulation of the high affinity IL-2 receptor subunit, IL-2Rα, our results suggest that selective IL-2Rα engagement uniquely promotes T cell persistence and anti-tumor efficacy in the absence of lymphodepletion. Our findings underscore the importance of T cell polarizing conditions in dictating the requirement of adoptively transferred T cells for exogenous cytokine.
Residual antigen presentation by CD40-licensed DCs instructs memory CD8+ T cells differentiation program to influenza (P1412)
Authors: Ballesteros-Tato André , León-Ruiz Beatriz , Lund Frances , Randall Troy 
Published: January 2023

Abstract
Here we show that residual antigen presentation by CD40-licensed dendritic cells (DCs) tunes memory CD8+ T cell responses to influenza. Primary and secondary CD8+ T cell responses to influenza nucleoprotein (NP) were reduced in Cd40-/- mice. When primed in Cd40-/- mice, NP-specific memory CD8+ T cells responded poorly to re-challenge, even in a CD40 sufficient environment. Defective programming in Cd40-/- mice was due to inefficient late cross-presentation of NP by CD70-expressing CD103-CD11bhi DCs. We demonstrated that residual antigen presentation by CD40 licensed CD70-expressing CD103-CD11bhi DCs prevented apoptosis of IL-2 responding CD8+ T cells and promoted memory CD8+ T cells differentiation by allowing combined TCR- CD25-CD27 signaling during the contraction phase of the primary response. We also show that CD40 signaling preferentially licensed CD70-expressing CD103-CD11bhi DCs to cross-present the more abundant influenza antigens. As a consequence, T cells of different specificities received different memory programing signals during the contraction phase depending on the antigen abundance, and the memory pool result enriched in T cell clones with a TCR repertoire selected for dominant epitopes. Taken together our results suggest that diferential residual antigen presentation by CD40 licensed CD70-expressing CD103-CD11bhi DCs represents a mechanism to enhance the fitness of the memory T cell responses
Polarized release and retroviral subversion of TCR-enriched microvesicles at the T cell immunological synapse (P6155)
Authors: Choudhuri Kaushik , Llodra Jaime , Tsai Jones , Roth Eric , Gordo Susana , Wucherpfennig Kai , Kam Lance , Stokes David , Dustin Michael 
Published: January 2023

Abstract
The recognition events that mediate adaptive cellular immunity depend on intercellular contacts between T cells and antigen presenting cells (APC). T cell signaling is initiated at these specialized junctions between T cells and APCs, known as the immunological synapse (IS), when surface-expressed antigen receptors (TCR) recognize peptide fragments of pathogens (pMHC) on APCs. Using high resolution optical and electron microscopy, optical-EM correlation, and electron-tomography, we show that centrally accumulated TCR at the IS is located on the surface of extracellular microvesicles that bud at the IS center and are recognized by APC bearing cognate antigen. An early endosomal-sorting complex required for transport (ESCRT) sorts TCR for inclusion in microvesicles, while terminal ESCRT components mediate scission of microvesicles from the T cell plasma membrane. The HIV polyprotein GAG co-opts this process by displacing TCR from microvesicles, resulting in release of virus-like particles at the antigen-dependent IS. We conclude that post-signaling TCR accumulates at the IS center and is released in extracellular microvesicles by an ESCRT-dependent mechanism triggered by T cell activation. These microvesicles mediate intercellular communication in immune cell collaboration, and can be co-opted by HIV GAG for viral transmission across antigen-dependent synapses.
Loss and aberrant function of NKp44+ ILCs in the SIV-infected gut are associated with RORγt suppression (P6175)
Authors: Reeves R. Keith , Li Haiying 
Published: January 2023

Abstract
Recent studies have identified a novel NK cell-like population found in mucosal tissues and mucosae-draining lymph nodes distinguishable by high surface expression of NKp44 and referred to as innate lymphoid cells (ILCs). ILCs secrete IL-17/IL-22 which are critical for epithelial homeostasis and thus could play a role in gut integrity, a major factor in HIV/SIV pathogenesis. During chronic SIV infection, numbers of ILCs declined ~8-fold in gastrointestinal tissues, but remained comparable to naïve animals in distal sites such as mesenteric lymph nodes and lungs. Loss of ILCs was associated with up to 100-fold increases in apoptosis. Interestingly, gut ILCs expressed NFIL3, a classic NK cell-related transcription factor, as well as the IL-17 transcriptional regulator, RORγt, but during SIV infection RORγt was significantly downregulated associated with decreased IL-17 secretion. In these same animals increased IFN-γ secretion and acquired cytotoxicity were associated with an increase in ILC NFIL3. Loss and functional skewing of ILCs in vivo correlated with upregulated IDO-1 and IFN-α in the gut, and in vitro treatment of ILCs with IDO-1 catabolites and IFN-α also suppressed RORγt expression, ablated IL-17 production and induced apoptosis. SIV infection results in massive numerical and functional depletion of ILCs in the inflamed gut, but not distal sites. This phenomenon was due, at least in part, to IDO-1- and IFN-α-induced apoptosis and suppression of RORγt.
Innate cytokines associate with exosomes in plasma of HIV-1+ individuals (P6205)
Authors: Johnson Kateena , Chu Jane , Huang Ming , Powell Michael , Villinger Francois , Bond Vincent 
Published: January 2023

Abstract
Purpose: Our group and others have demonstrated a potential role for exosomes in HIV pathogenesis. Exosomes secreted from HIV-infected cells were shown to contain coreceptors for HIV. We have shown HIV Nef-containing exosomes from HIV-infected cells induced apoptosis in bystander CD4+ T cells. This suggests that exosomes from HIV-infected cells could drive CD4 depletion and immune activation contributing to HIV pathogenesis. Thus, exosomes from HIV-1+ viremic individuals were investigated for protein content. Methods: Exosomes were isolated from plasma of ARV naïve, HIV+ and negative control individuals by differential centrifugation followed by iodixanol density gradient ultracentrifugation. Biochemical characterization included AChE activity, flotation in iodixanol gradients, and Western analysis for exosomal markers. Plasma and purified exosomes were also tested for levels of 30 cytokines and chemokines by multiplex assay. Results: The concentrations of 10 innate cytokines (IL-15, IP-10, FasL, CD40L, IFNα, IFNβ, MIP-1α, MIP-1β, TNFα, and ICAM-1) were found to be associated with and significantly elevated in exosomes from HIV+ individuals relative to negative controls, suggesting a contributing role in generalized chronic immune inflammation. Conclusion: This is the first demonstration of innate cytokines in exosomes of HIV+ individuals. It suggests a potential mechanism for HIV-1 driven immune modulation and propagation of HIV via activation of bystander target cells.
Cell-intrinsic HIV-1 immune responses in conventional dendritic cells from HIV-1 elite controllers (P6171)
Authors: Martin-Gayo Enrique , Hickman Taylor , Pimenova Dina , Pereyra Florencia , Rosenberg Eric , Lichterfeld Mathias , Yu Xu 
Published: January 2023

Abstract
Introduction: Recent data suggest that in most HIV-1-infected individuals, cell-intrinsic immune responses of conventional dendritic cells (cDC) are blocked by the host proteins Samhd1 and Trex1. Elite controllers (EC) control HIV-1 replication in the absence of treatment, but immune defense mechanisms in these patients are not well understood. Here, we investigated cell-intrinsic immune responses to HIV-1 in these specific patients. Methods: PBMC from EC, untreated chronic progressors (CP), HAART-treated and HIV-1 negative subjects were ex vivo infected with HIV-1. Expression of viral replication products, type I interferons, Samhd1 and Trex1 were analyzed by qPCR. Results: cDC from HIV-1 negative persons were moderately susceptible to HIV-1, while cDC from EC and CP supported HIV-1 replication very weakly. However, in CP, HIV-1 replication was blocked at the level of early reverse transcription, likely as a result of high-level Samdh1 expression, while In EC, reverse transcription was unaltered, and restriction of viral replication mostly occurred at the level of integration. Functionally, these altered patterns of viral restriction in cDC from EC were associated with increased cellular activation, secretion of type I interferons and improved abilities to prime T cell responses. Conclusion: cDC from EC can mount cell-intrinsic immune responses against HIV-1, which may support the generation of highly effective HIV-1-specific T cell responses in these patients.
Lysine dimethylation in the BH4 domain of Bcl-2 and functional significance (P1306)
Authors: Li Wenqing , Xiao Zhen , Ravichandran Sarangan , Collins Jack , Veenstra Timothy , Durum Scott 
Published: January 2023

Abstract
Bcl-2 is an anti-apoptotic protein that can protect developing and mature T cells against a variety of apoptotic signals. The Bcl-2 family of proteins, compromised of anti-apoptotic as well as pro-apoptotic members, plays key roles in apoptosis; the balance of these members determines cell survival. Anti-apoptotic Bcl-2 contains four conserved Bcl-2 homology (BH) domains and the BH4 domain is indispensable for its ability to suppress cell death. Using mass spectroscopy to analyze a murine T cell line, we identified dimethylation of two lysine residues in the BH4 domain: K17 and K22. Mutation of K22 (but not K17) to alanine abrogated Bcl-2 binding to pro-apoptotic proteins Bax, Bak, Bad, Bim and Puma and greatly reduced Bcl-2 anti-apoptotic function in cytokine withdrawal. This assigns functional importance to K22 (which is conserved across species and other anti-apoptotic members) and suggests that dimethylation could affect its function. K22, based on structural models, appears to be involved in stabilizing peptide secondary structure, rather than directly interacting with pro-apoptotic proteins. We developed a monoclonal antibody that recognizes dimethylated K22 in the Bcl-2 sequence and detected expression in murine B cells (but not thymocytes). This suggests that Bcl-2 dimethylation may regulate B cell survival.
CD56 is a legitimate immune receptor regulating T cell and NK cell effector function, and its expression level predicts successful aging (P1324)
Authors: de Vallejo Abbe , Griffin Patricia , Montag David , Nussbaum Ryan , Gunawardema Naomi , Studenski Stephanie 
Published: January 2023

Abstract
CD56 is a classical marker of NK cells, but its direct relevance to the elaboration of NK cell effector function is not known. In two cohort studies of aging, we have found that high levels of CD56 expression on both NK cells and T cells are highly correlated with unimpaired cognitive and physical ability among older adults aged ≥65 years. In multivariate analysis of collated data on clinical and laboratory assessments of health and performance, we identified CD56 as a key immunologic predictor of successful aging, an emerging physiologic and clinical construct pertaining to the maintenance of functional independence in old age despite a history of disease and concurrent comorbid conditions. In biological assays using primary T and NK cells, and various immortalized T/NK cell lines, ligation of CD56 alone was sufficient to induce activation of NK cells, and of T cells in a TCR-independent manner. CD56 triggering effectively elicited expression of activation antigens such as CD69 and CD25, and the production of various cytokines including IL-2, IL-4, and IFN-γ. CD56-mediated activation of both T cells and NK cells involved LAT phosphorylation. Clearly, CD56 is a functional immune receptor capable of effecting proximal and distal cellular outcomes. TCR-independent CD56-driven activation of T cells in particular is consistent with the concept of beneficial immune remodeling that has been proposed to underlie healthy aging and long-term survivorship.