bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sepsis-Induced T Cell Immunoparalysis: The Ins and Outs of Impaired T Cell Immunity
Authors: Jensen Isaac J. , Sjaastad Frances V. , Griffith Thomas S. , Badovinac Vladimir P. 
Published: January 2023

Abstract
Sepsis results in a deluge of pro- and anti-inflammatory cytokines, leading to lymphopenia and chronic immunoparalysis. Sepsis-induced long-lasting immunoparalysis is defined, in part, by impaired CD4 and CD8 αβ T cell responses in the postseptic environment. The dysfunction in T cell immunity affects naive, effector, and memory T cells and is not restricted to classical αβ T cells. Although sepsis-induced severe and transient lymphopenia is a contributory factor to diminished T cell immunity, T cell–intrinsic and -extrinsic factors/mechanisms also contribute to impaired T cell function. In this review, we summarize the current knowledge of how sepsis quantitatively and qualitatively impairs CD4 and CD8 T cell immunity of classical and nonclassical T cell subsets and discuss current therapeutic approaches being developed to boost the recovery of T cell immunity postsepsis induction.
Morphine and TAT: Additive modulation of wound healing events (100.15)
Authors: Martin Josephine L. , Roy Sabita 
Published: January 2023

Abstract
Interactions between chronic morphine and the HIV-1 protein TAT are believed to play a major role in accelerating wound pathogenesis of HIV positive patients. Chronic morphine users infected with HIV often display infected and improper wound closure events compared to non-drug abusive HIV populations. Although it is well documented that HIV-1 proteins such as TAT up-regulate chemokine secretion, our laboratory is the first to show morphine suppression of TAT-induced leukocyte migration and chemokine secretion. Simultaneous treatment of morphine and lipopolysaccharide results in altered cytokine profiles of monocyte chemotactic protein-1 and macrophage inflammatory protein-2. Such alterations lead towards altered immune cell recruitment profiles of neutrophils and macrophages resulting in inadequate bacterial clearance and ultimately, delayed closure of wounds.
We have recently found further exacerbation of murine wound beds following co-administration of Tat proteins and morphine using ELISA, flow cytometry and histological assays.
Taken together, our data implicates a mu opioid receptor-mediated immune suppression of wound healing events following Tat and chronic morphine treatment and shows the potentially detrimental effects chronic morphine administration may have in the wound healing processes of HIV positive patients.
Supported by: NIDA R01-DA12104, K02-DA015349 and P50-DA11806.
Wide Expression and Cellular Localization of Inflammation-associated TSG-6 Protein in Atherosclerotic Lesions (100.17)
Authors: Mindrescu Catalin , Coppola John , Doty Stephen , Antonescu Cristina , Staniloae Cezar 
Published: January 2023

Abstract
TNF-stimulated gene 6 (TSG-6) is induced by TNF-α during inflammation and its secreted product TSG-6 glycoprotein is involved in immune-mediated inflammatory diseases and fertility. It regulates COX-2 and prostaglandin synthesis, and participates in extracellular matrix remodeling. Considering the chronic inflammatory nature of atherosclerosis we hypothesized that TSG-6 is expressed in atherosclerotic plaques and investigated TSG-6 protein expression and cellular distribution on 12 superficial femoral artery endarterectomy specimens from 6 diabetic and 6 non-diabetic patients with peripheral vascular disease. Six histologically normal radial artery specimens were analyzed as control. Paraffin embedded samples were studied by immunohistochemistry using a goat polyclonal anti-human-TSG-6 antibody. TSG-6 expression was consistently present in all atherectomy specimens but not in control specimens. A distinct, strong cytoplasmic staining pattern was uniformly detected in the endothelial lining of the intima, as well as in the neo-vessel proliferation of the plaque. Cytoplasmic staining was also identified in the smooth muscle cell proliferation of the neo-intima. Patchy TSG-6 expression was noted in the extracellular matrix. Within the inflammatory plaques from diabetic patients, TSG-6 stained the foamy macrophages. Our study identifies for the first time the preferential expression of TSG-6 in atherosclerotic lesions and characterizes its distribution within the cellular and matrix components of the plaque. TSG-6 is a novel inflammatory mediator of atherosclerosis and a potentially new marker of endothelial / smooth muscle cell activation.
High levels of ROS block the differentiation of human and mouse osteoclasts (100.5)
Authors: Moreno Jose Luis , Celluzzi Christina M , Keegan Achsah D , Williams Mark S 
Published: January 2023

Abstract
During chronic inflammation at skeletal sites, production of inflammatory cytokines by cells from the joint tissues and recruited activated T cells generate a pro-inflammatory environment with high levels of Reactive Oxygen Species (ROS). A critical cytokine expressed by the joint tissue and by the activated T cells is RANKL, which exerts its activity by inducing the differentiation of osteoclasts (OC) from cells of the monocyte-macrophage lineage. The effect of RANKL on the OC is mediated by a transient generation of ROS which are critical in regulating RANKL signaling. Therefore, we investigated the effect of increased ROS levels on OC differentiation from human and mouse progenitors. We found that addition of the glutathione depleting drug BSO to the cultures blocked OC generation from CD14+ human PBMC and from mouse bone marrow cells. This blockage was reversible; withdrawal of BSO allowed for the recovery of OC formation. The level of recovery diminished with increasing concentrations of BSO. The inhibition was time dependent, and it had a more profound effect if monocytes were the precursor cells rather than macrophages. Thus, while RANKL-induced OC development requires ROS, excess ROS can inhibit OC development.
(Supported by PHS AI59775 (ADK) AI49807 (MSW))
Toll-Like Receptor Expression is Necessary for Synovial Type II Cells to Respond to LPS: a Potential Mechanism of Reactive Arthritis (101.14)
Authors: Hao Hsiao-Nan , Yang Shang-Yu , Wooley Paul H 
Published: January 2023

Abstract
Reactive arthritis is predominantly associated with abnormal immune reactivity to gram-negative bacterial infections in children and young adults, but the precise molecular mechanism of the pathology remains unknown. Our previous results indicated fibroblast-like synovial cells (FLSC) express inducible Toll-like receptors (TLR). In this study we investigated the potential relationship of synovial cell TLR-dependent responses to the pathologen of reactive arthritis. To define TLR positive synovial cell subtypes, cell specific surface markers and flow cytometry were used both to isolate and to characterize the cells from mouse synovial tissues. Using synovial cell cultures and anti-TLR oligonucleotides, we examined whether the synovial immunological responses to LPS is dependent on TLR expression. Our data shows partial of FLSC were consecutive positive for TLR4, and inducible by LPS or cytokines. These cells were different from typical fibroblasts which are TLR4 negative. The TLR4+ FLSCs contribute approximately thirty percent of the isolated FLSC population, and sensitive to gram-negative bacterial endotoxin to produce inflammatory cytokines. Suppression of TLR4 expression on the cell surface significantly decreased the production of LPS-induced cytokines. The findings suggest synovial cells may orchestrate the immunological responses to bacterial components via TLR-based mechanisms.
MAP kinase phosphatase-1 plays a pivotal role in the protective effects of dexamethasone against endotoxic shock (101.15)
Authors: Wang Xianxi , Nelin Leif D , Kuhlman Joshua R , Meng Xiaomei , Welty Stephen E , Liu Yusen 
Published: January 2023

Abstract
Although glucocorticoids potently inhibit the biosynthesis of pro-inflammatory cytokines, the underlying mechanism is still not fully understood. We have previously shown that glucocorticoids induce the expression of MAP kinase phosphatase (MKP)a-1 in innate immune cells. Since MKP-1 is a critical negative regulator in innate immune response, we hypothesize that induction of MKP-1 plays an important role in the inhibition of pro-inflammatory cytokine biosynthesis by glucocorticoids. To test this hypothesis, we compared the anti-inflammatory effects of dexamethasone between wild-type and Mkp-1-deficient mice. We found that prophylactic administration of dexamethasone completely protected the wild-type mice, but not the Mkp-1−/− mice, from mortality caused by high-dose of lipopolysaccharides (LPS). Dexamethasone attenuated TNF-α production to lower levels in LPS-challenged wild-type mice than in similarly challenged Mkp-1−/− mice. Likewise, TNF-α and IL-6 production in vitro by wild-type macrophages upon LPS stimulation was decreased to lower levels by dexamethasone than did cytokine production by Mkp-1−/− macrophages. Dexamethasone pretreatment shortened the duration of p38 and JNK activities in LPS-stimulated wild-type macrophages, but had little effect in similarly treated Mkp-1−/− macrophages. Our results indicate that the inhibition of p38 and JNK activities by glucocorticoids is mediated by enhancing MKP-1 expression. These results demonstrate that induction of MKP-1 constitutes an important part of the anti-inflammatory mechanism of glucocorticoids.
Gene delivery strategies for targeting microglia cells (101.17)
Authors: Puzis Leopold , Odashima Kay , Pearse Damien D. 
Published: January 2023

Abstract
Microglia (MG) are the resident immune cells of the CNS and are implicated as both important sensors for biological defense as well as mediators of various pathological conditions through the various inflammatory and oxidative molecules they produce. In the current study we compared different techniques for gene targeting of microglia using non-viral and viral methods in vitro for eventual application to the in vivo setting. For this investigation we employed the EOC2 MG cell line and three methods of gene introduction for pmax plasmid-GFP:
electroporation,cationic lipid co-delivery andlentiviral vector infection.
Electroporation when employed with various buffers and at different voltages led to low transfection efficacies (&gt;1%). Transfection with cationic lipids (LipofectAmine, NovaFector, GeneFector) showed higher transfection rates (%), but these were still relatively low. In contrast, lentiviral vector delivery produced robust infection (&gt;95%) and GFP expression. In addition, the activation of the cells, so that they underwent morphological (rounded) and immunophenotypical (ED1 immunoreactivity) changes, prior to or during transfection with the lower efficiency methods, did dramatically enhance efficiency (&gt;95%). We conclude, that in CNS pathologies, where MG cells are activated, both non-viral and viral methods of gene therapy will likely be highly successful for the introduction of specific anti-inflammatory or immune suppressive genes.
FUNDING: The Miami Project to Cure Paralysis, The Buoniconti Fund, The US Army Medical Research and Material Command
Biliary obstruction expands TREG which suppress liver T cell function (101.2)
Authors: Katz Steven C , Chaudhry Umer , Kingham T. Peter , Raab Jesse , DeMatteo Ronald P 
Published: January 2023

Abstract
INTRODUCTION: The mechanism by which obstructive jaundice predisposes to infection has not been clearly defined. We hypothesized that extrahepatic biliary obstruction would directly impair the function of T cells within the liver.
METHODS: Bile duct ligation (BDL) or sham laparotomy (SL) was performed in mice. Liver or spleen T cell function was assessed by proliferation in response to dendritic cells or anti-CD3 stimulation. TREG (CD4+CD25+FoxP3+) were measured by flow cytometry. CD4+C25+ (70% FoxP3+) liver T cells were purified by FACS and used as suppressors.
RESULTS: Following BDL, liver T cells were markedly less responsive to allogeneic or antigen-loaded syngeneic dendritic cells. In contrast, spleen T cell function was preserved. Even in the presence of direct anti-CD3 stimulation, liver T cells from BDL mice were functionally deficient and produced less IL-2 relative to spleen T cells. BDL expanded the number of liver TREG which suppressed the response of CD4+C25− T cells to DC in vitro.
CONCLUSION: BDL expands TREG, which may contribute to the increased risk of infection in patients with obstructive jaundice.
The effects of Immunosuppressive agents on NK cell function (102.18)
Authors: Wai Lu-En , Fujiki Masato , Takeda Saori , Hsieh Christine , Martinez Olivia M , Krams Sheri M 
Published: January 2023

Abstract
Natural killer (NK) cells are potent effector cells in allogeneic bone marrow and xeno transplantation. NK cells infiltrate solid organ allografts and a role for NK cells in acute and chronic rejection has been suggested. Little is known about the effects of immunosuppressive agents on NK cells
Primary NK cell lines were analyzed for proliferation in the absence or presence of increasing doses (0–100 ng/ml) of cyclosporine (CSA), FK506 (FK) or Rapamycin, (RAPA). NK cells demonstrated robust proliferation both in the absence and presence of CSA and FK. In contrast, RAPA significantly (p&lt; 0.05) inhibited the proliferation of NK cells in a dose-dependent manner. The effects of immunosuppressive agents on NK cells were determined in a rat liver transplant model. Peripheral NK cell levels in graft recipients treated with CSA and FK remained stable whereas there was a marked decrease in the levels of NK cells in recipients treated with RAPA. The immunosuppressive agents did not alter IFNγ production by NK cells however NK cell–mediated cytotoxicity was significantly (p&lt; 0.05) inhibited in the presence of RAPA (but not CSA or FK).
These data indicate that CSA and FK have little effect on NK cell function. In contrast, NK cell proliferation and function are significantly decreased in the presence of RAPA. Understanding the differential effects of immunosuppressive drugs on NK cell function is important in clinical transplantation.
Diversity and fate of pre-immune anti-tumor T cells after umbilical cord blood transplantation (102.28)
Authors: Merindol Natacha , Caty Martine , Gimmig Sylvain , Duval Michel , Champagne Martin A. , Soudeyns Hugo 
Published: January 2023

Abstract
Umbilical cord blood (UCB) has been used successfully as a source of progenitor cells to treat a variety of haematological and immune disorders in children. Donor-derived T cells play a key role in antitumoral and antiviral immunity and contribute to the survival of the graft recipient. Here we studied the diversity, persistence, fate and antitumoral function of UCB-derived T cells and characterized their persistence and function in the graft recipient. Naïve CD8+ T cells specific for the Melan-A26-35 peptide were isolated from UCB and UCB recipients at 1, 3, and 6 months post-transplant using Melan-A26-35-HLA-A2 tetramers and cell sorting, and were cultured in the presence of IL-2, IL-7, PHA and peptide. Melan-A-specific cytotoxicity was tested in 51Cr-release assays, and the clonal diversity of the pre-immune Melan-A-specific repertoire was determined by sequencing of the T cell receptor β chain CDR3 region. 14 UCB recipients were enrolled. Melan-A-specific CD8+ T cells were detected at a median frequency of 2.28% (range=0.0–8.18%) in 4 of 6 UCB samples initially analyzed. These cells proliferated in the presence of Melan-A peptide in 4 of 6 cases, and showed cognate cytotoxicity in 2 of 6 cases. Finally, tetramer-positive cells were detected in UCB recipients as early as 1 month post-transplant. This study will lead to a better understanding of the fate of the pre-immune T cell repertoire in UCB recipients.
A Novel In Vitro Murine Epithelial Culture System To Study Cutaneous GVHD (102.6)
Authors: Arditti Fabian David , Friedman Thea M , Murphy George , Korngold Robert 
Published: January 2023

Abstract
Graft-versus-host disease (GVHD), following allogeneic hematopoietic stem cell transplantation, involves a donor T cell attack against a number of tissue systems, including the skin, resulting in high levels of morbidity and mortality. The development of an in vitro murine model system for examining the underlying processes of specific target cell injury would be invaluable for increasing the understanding of the immunobiology of GVHD. In an effort to mimic the in situ nature of the target tissue, primary skin epithelial cell cultures were generated from ear skin of adult mice, after aggressive enzymatic digestion, without addition of growth factors or chemokines. The cultured cytokeratin-positive epithelial cells can be expanded and cryo-preserved for several passages. To test the ability of alloreactive T cells to mount responses against cultured epithelial cells, T lymphocytes from C57Bl/6 (B6) mice were added to the epithelial cells grown from minor histocompatibility antigen-mismatched BALB.B or syngeneic B6 mice. In vitro responses were characterized by day 6 supernatant interferon gamma levels (anti-BALB.B, 336.5 ± 32 pg/ml; anti-B6, 16.7 ± 12.2 pg/ml). Up-regulation of H2Kb was also observed on the surface of BALB.B epithelial cells (3.5 ± 2.2 and 90.5 ± 25 MFI on days 1 and 7, respectively). In addition, there was an increased amount of epithelial cell death, as denoted by 7-AAD staining.
Reduced sensitivity of effectors to regulation, and not reduced Treg function, may lead to autoimmune diabetes in NOD (128.1)
Authors: Nettles Adam , Honkanen-Scott Minna , Bansal-Pakala Pratima 
Published: January 2023

Abstract
T regulatory cells [Tregs] play an important role in controlling autoimmunity. The NOD mouse provides an excellent model to delineate the role of Tregs. Transfer experiments have shown that Tregs can prevent the progression of T1D. Similar results were obtained from depletion of Tregs in vivo. However, quantitative and qualitative changes in Tregs with the progression of T1D have not been examined in detail. In this study we studied changes in number and function of Tregs during progression of T1D in NOD. Since NOD males and females are very similar, yet the incidence of diabetes in males is much lower than the females, we examined sex based differences in Tregs and effectors.
We examined the expression of CD4+CD25+FoxP3+ to define the quantitative differences in Tregs. Time-course analysis showed that Treg numbers remained constant or slightly increased from 6 week to 16 weeks of age. Also, both female and male NOD had similar numbers of Tregs, suggesting that T1D progression is not due to a quantitative change in Tregs. Functional assays for purified Tregs from both male and female NOD showed marginal differences between their activities in vitro and in vivo. We found that islet-reactive T cells from female NOD were more auto-aggressive and refractory to regulation by Tregs than male effectors. These data suggest, for the first time, that refractory effector T cells, and not the number or function of Tregs, may account for T1D.
Humoral and cellular immune responses to IGF-2 in type 1 diabetes (128.21)
Authors: Louis Céline , Renard Chantal , Bosseloir Alain , Weets Ilse , Gorus Frans , Geenen Vincent 
Published: January 2023

Abstract
Background
IGF-2 is the dominant member of insulin family expressed in the thymus, Igf2 expression contributes to complete tolerance of insulin, and Igf2 transcription is defective in the thymus of diabetes-prone BB rats. Objectives
To evaluate the immunological tolerance to IGF-2 in T1D patients; andTo analyse the type of cellular response induced by IGF-2 B11-25, the homologous sequence of Insulin B9-23, a
major T1D auto-antigen.
Results
Using a sensitive and specific radio-binding assay, the presence of autoantibodies against IGF-2 was investigated in 100 newly diagnosed T1D patients and 100 healthy controls. No significant difference appeared between the two groups.We also investigated the profile of IL-10/IFN-γ secretion following presentation of Insulin B9-23 and IGF-2 B11-25
to PBMCs from T1D patients and healthy controls. Compared to Insulin B9-23 in ELISpot, ELISA and real-time RT-PCR, the presentation of IGF-2 B11-25 induced a regulatory/tolerogenic profile with a significantly higher increase of IL10 transcription and IL-10 secretion.
Conclusions
The very strong tolerance to IGF-2 is associated to active regulatory properties that might be exploited for development of a tolerogenic self-vaccine against T1D.
(Supported by the Belgian NFSR, Walloon Region TOLEDIAB project, and FP6 Euro-Thymaide Integrated Project)
Regulatory interactions between two innate immune cells - iNKT and MZB cells - suppress autoimmunity (128.24)
Authors: Wen Xiangshu , Yang Jun-Qi , Singh Ram Raj 
Published: January 2023

Abstract
In vivo activation of iNKT cells prevents autoimmune diseases such as lupus at least during early stages of disease development. Conversely, loss of iNKT cells in CD1d-KO mice worsens nephritis in lupus-prone BWF1 mice. Mechanisms of regulation of autoimmunity by iNKT cells remain poorly understood. Here, using a variety of in vitro and in vivo experimental systems, including adoptive transfers in SCID and iNKT cell-deficient (Ja18–/–) mice, we demonstrate that although iNKT cells trans-activate B cells to increase expression of activation markers and production of normal Ig, they selectively suppress IgG autoantibody production. Whereas increased expression of B cell activation markers occurs via cytokines secreted by iNKT cells, iNKT cell-mediated suppression of IgG autoantibody production requires a direct contact between iNKT cells and B cells. Furthermore, although iNKT cells increase production of cytokines by other immune cells, they specifically inhibit IL-10 production by B cells in a contact-dependent manner. Furthermore, whereas iNKT cells increase activation markers on all B cell subsets including B1, B2 and marginal zone B (MZB) cells, they significantly reduce the MZB cell population in lupus-prone BWF1 mice. Further support for the role of iNKT cells in regulating MZB cells came from iNKT cell transgenic (Va14Tg) and deficient (Ja18–/–) mice. Whereas the proportion of MZ-B cells are increased in Ja18–/– mice as compared to wild type BALB/c mice, these cells are reduced in Va14Tg mice. Thus, iNKT cells suppress autoreactive B cells and prevent systemic autoimmunity. They do so via regulating MZ-B cells. Ongoing investigations will elucidate how activated iNKT cells activate MZB cells but reduce their IL-10 and autoantibody production.
Harnessing the Fas pathway for type 1 diabetes therapy (128.29)
Authors: Mohamood Abdiaziz Sheikh , Xiao Zuoxiang , Schneck Jonathan P , Hamad Abdel Rahim A 
Published: January 2023

Abstract
The Fas pathway is not required for T cell activation yet loss-of-function mutation that inactivates Fas (lpr) or FasL (gld) prevents autoimmune diabetes in NOD mice without interfering with normal immune responses. However, the Fas pathway has not been considered a viable therapeutic option because these mutations also cause double negative (DN) T cell lymphoproliferation and lupus-like syndrome. Using bone marrow chimeras and adoptive transfer, we show that gld mutation on either hemopoietic or nonhemopoietic compartment is equally protective from autoimmune diabetes; however, gld mutation on hemopoietic cells is responsible for most of DN T cell lymphoproliferation. Therefore, limiting gld mutation to the nonhemopoietic tissue completely controls diabetogenicity of wild type T cells without causing lymphoproliferation. The protection is recapitulated therapeutically by treating NOD-wt mice with FasL neutralizing antibody. Anti-FasL treatment curtailed insulitis and did not lead to lupus-like disease or DN T cell lymphoproliferation. These results suggest the potential of generating a new class of immunotherapy that is based on targeting T cell apoptosis inducing ligand rather than costimulatory pathways.