bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tonsil-derived stromal cells reduces CCl4-induced liver fibrosis in mice via autophagy activation (P2180)
Authors: Woo So-Youn , Park Minhwa , Lee Hye Jin , Kim Ji-Yon , Ryu Kyung-Ha 
Published: January 2023

Abstract
Liver transplantation is the treatment of choice for chronic liver failure due to fibrosis, however, it faces several difficulties, including donor shortage, surgery-related complications, immunological rejection, and high medical cost. Cell transplantation therapy would be the minimally invasive and alternative methods, potentially with fewer complications. As cell source, we isolated mesenchymal stromal cells from human palatine tonsils (T-MSCs) and injected into CCl4-induced liver injury model of the mice. We found that T-MSCs were differentiated into hepatocyte-like cells under 25 ng/ml insulin-like growth factor, 25 ng/ml hepatocyte growth factor, 10-7 M dexamethasone, and 10 ng/ml oncostatin M for three weeks in vitro. After injecting into CCl4-liver damaged mice, T-MSCs were differentiated into albumin positive cells but were not detected in the group of mice with normal liver. During this process, T-MSCs showed accumulation of autophagosome-bound LC3B, upregulation of proautophagic beclin-1, and ubiquitin positive in vivo. Our data imply that T-MSCs contributed to reduction of CCl4-induced liver fibrosis through autophagy activation pathway.
Nonmyeloablative conditioning employing busulfan and sirolimus permits donor-dominant stable chimerism in a murine model of haploidentical allogeneic stem cell transplantation (P2192)
Authors: Weitzel R. , Fitzhugh Courtney , Phang Oswald , Hsieh Matthew , Tisdale John 
Published: January 2023

Abstract
Allogeneic stem cell transplant (SCT) is a potentially curative option for many hematologic diseases, yet donor availability remains a challenge, and haploidentical SCT is associated with a higher risk of rejection. We successfully translated a model based on total-body irradiation (TBI) with to humans; however, TBI involves toxicities and risk of malignant side-effects, and we hypothesize that busulfan may be a superior alternative in some settings. G-CSF-mobilized F1/J (C57BL/6-BALB/c hybrid) splenocytes were injected into C57BL/6 hosts who had received 10-30mg/kg IP busulfan the previous day. All recipients received 31 days of 3mg/kg IP sirolimus beginning the day before transplant. No GvHD was observed and counts recovered by week 8 in all groups. Mice conditioned with 30mg/kg busulfan displayed remarkably high donor chimerism (&gt;90%), which was significantly higher than 300cGy TBI-conditioned control mice (64-86%) across all timepoints. Mice receiving 30mg/kg or 20mg/kg busulfan maintained stable chimerism for over 18 months. Mice conditioned with 10mg/kg busulfan showed only transient chimerism through 4 weeks. Proliferation of CD4+ cells in response to donor increases with decreasing busulfan, and mice receiving 30mg/kg responded more strongly than mice receiving TBI. A memory effect was observed in mice that received 10mg/kg busulfan and lost their grafts. Notably, a slightly higher incidence of host-derived Tregs in mice was also observed after graft rejection.
Development of engineered heart tissue from MHC-I downregulated non-cardiac fibroblasts and stem cell derived cardiomyocytes. (P2224)
Authors: Karabekian Zaruhi , Ding Hao , Jamshidi Aria , Sarvazyan Narine 
Published: January 2023

Abstract
Our goal was to test a hypothesis that downregulation of MHC class I molecules via inhibition of beta2-microglobulin gene expression can minimize immunogenicity of non-cardiac fibroblasts and stem cell derived cardiomyocytes without impairing ability of these cells to form functional cardiac tissue. When co-cultured with mouse stem cell cardiomyocytes, two different types of non-cardiac mouse embryonic fibroblasts (NIH3T3 and WT19) supported the development of three-dimensional cardiac fibers. Fibroblasts had to be irradiated to ensure optimal growth, so that they did not overgrow non-replicating cardiomyocytes and a proper ratio of fibroblasts to cardiomyocytes was needed to create optimal fibers. The fibers also formed when both cell types underwent downregulation of MHC-I molecules. The latter was achieved by inhibition of beta2-microglobulin gene expression using shRNA technology and was confirmed by qPCR and FACS analyses. The downregulation of MHC-I molecules made fibroblasts and cardiomyocytes less susceptible to T-cell recognition in both in vitro and in vivo experiments. We conclude that readily obtainable non-cardiac fibroblasts can be used to engineer functional cardiac tissue and that downregulation of MHC-I can make such tissue less prone to immune rejection.
Inhibition of CD1d-mediated antigen presentation by the TGF-β/Smad signaling pathway (APP3P.113)
Authors: Iyer Abhirami Kannan , Bailey Jennifer C. , Gourapura Renukaradhya , Lin Yinling , Nguyen Hoa B. , Brutkiewicz Randy 
Published: January 2023

Abstract
CD1d is a MHC class I-like molecule that presents lipid Ags to a subpopulation of innate lymphocytes called natural killer T cells. In a prior study, we showed that the mitogen-activated protein kinase p38 is a negative regulator of Ag presentation by CD1d. It is unknown how p38 is activated to induce this regulation; however, several studies have implicated a role for the immunoregulatory cytokine, TGF-β, in the activation of p38. Therefore, we hypothesized that TGF-β negatively regulates the CD1d-mediated Ag presentation pathway. Indeed, a dose-dependent decrease in CD1d-mediated Ag presentation and impairment of lipid Ag processing was observed in TGF-β-treated antigen presenting cells. However, this inhibition was not through p38 activation. Instead, by employing a lentiviral shRNA-based approach, we found that the canonical TGF-β signaling elements, Smads 2, 3 and 4, contributed to the TGF-β-mediated inhibition of Ag presentation by CD1d. Furthermore, a side-by-side comparative analysis of the effect of TGF-β on CD1d and MHC class II-mediated Ag presentation revealed that in contrast to that observed with CD1d, TGF-β actually enhances MHC class II-mediated Ag presentation. Overall, these results suggest that the canonical TGF-β/Smad pathway has distinctly different effects on the regulation of CD1d- and MHC class II-mediated antigen presentation pathways.
HLA-DM and HLA-DO: bi-directional regulation of function, tuned by pH (APP3P.103)
Authors: Jiang Wei , Strohman Michael , Wang Nan , Hou Tieying , Mellins Elizabeth 
Published: January 2023

Abstract
Peptide processing and loading onto MHC-II in endosomes occurs across a wide pH range. We studied pH effects on DM/DO regulation of peptide loading in vitro, using soluble, recombinant molecules, including a stabilized mutant DO dimer (sDOAP11A) that inhibits DM normally. We tested different DO:DM ratios at pH 4.6-7.2 during loading of 4 HLA alleles. DO alone had no effect. DM inhibition was diminished at lower DO:DM ratios, implying stable DM association is required for DO effect. Inhibition occurred across the pH range, but was reduced at pH 4.6. Using a narrower pH gradient (4.56-5.30), across which DM function is equivalent, we identified DO as the primary target of pH effects. DO activity was reduced after pre-incubation at pH 5.4, and longer exposure, lower pH and higher temperature enhanced this effect. FRET analysis of DM/DO at low pH was consistent with DO release rather than conformational change of intact complexes. SPR, bio-layer inferometry and ELISA assays showed reduced DM/DO association at pH≤4.85, suggesting released DO binds back inefficiently. With pre-incubation at pH 4.6 or 5.4, DM-DOwt complexes acquired DM-like activity; analysis of active SEC fractions indicated presence of free DM. Pre-incubation of sDM alone at these pHs caused aggregation. Complexes of full-length DM and DOwt showed similar behavior to soluble molecules. We conclude that low pH generates free DM by releasing DO, which both inhibits and chaperones DM.
Potentiation of antigen-specific CTL generation by caTLR4 gene transduction into leukemic plasmacytoid dendritic cell line (PMDC11) (APP3P.108)
Authors: Takahashi Masuhiro , Iwabuchi Minami , Satoh Naoya , Uchiyama Takayoshi , Iwaya Shunpei , Oiwa Rie , Nishizawa Yoshinori , Hashimoto Shigeo , Bonehill Aude , Kasahara Noriyuki , Narita Miwako 
Published: January 2023

Abstract
Leukemic plasmacytoid dendritic cell (pDC) line with HLA-A02/24, PMDC05, which was established in our laboratory, acquired a higher antigen presenting ability by being transduced with lentiviruses encoding the CD80 gene. CD80 gene-transduced PMDC05 was named PMDC11. In order to establish a more potent antigen presenting cells for cellular immunotherapy in tumors, PMDC11 cells were transduced with a constitutively active (ca) TLR4 gene by using the Tet-On system (caTLR4-PMDC11). CD8+ T cells from healthy donors with HLA-A24 were co-cultured with WT1 peptide-pulsed caTLR4-PMDC11 cells. Priming with WT1 peptide-pulsed caTLR4-PMDC11 cells was performed every week. Immediately before the priming with caTLR4-PMDC11 cells, cultured cells were analyzed for the percentage and number of CD8+/WT1 tetramer+ T cells. CD8+/WT1 tetramer+ T cells were generated after 2-3 weeks of culture and much increased in caTLR4-PMDC11-primed CD8+ T cell culture compared with parental PMDC11-primed CD8+ T cell culture. These CD8+ T cells co-cultured with caTLR4-PMDC11 cells were demonstrated to be cytotoxic to WT1-expressing HLA-A24+ target cells. These data revealed that the antigen-specific cytotoxic T lymphocyte (CTL) inducing ability of PMDC11 was enhanced by being transduced with caTLR4 gene, and that caTLR4-PMDC11 cells could be applied as potent antigen presenting cells for generating antigen-specific CTLs in adoptive cellular immunotherapy against tumors and severe viral infection.
Human peripheral invariant natural killer T are adaptive lymphocytes, whose TCR CDR3β regulates their antigen specificity and reactivity to avoid harmful autoimmune responses (BA8P.124)
Authors: Chamoto Kenji , Imataki Osamu , Guo Tingxi , Saito Toshiki , Saito Akiko , Elia Andrew , Chiu Priscilla , Butler Marcus , Hirano Naoto 
Published: January 2023

Abstract
Human invariant natural killer T (iNKT) cells carry a sole variable element, complementarity determining region 3β (CDR3β) in their T cell receptors (TCRs). It has been widely accepted that iNKT cells recognize foreign and self-ligands via CD1d in a pattern recognition-like innate-type manner. Here, we have created a thymically unselected human iNKT Vβ11 TCR repertoire and isolated a series of iNKT TCRs with ultra-high to low autoreactivity. We have found three distinctive CDR3β sequence motifs that dictate high iNKT TCR autoreactivity. In vitro and in vivo studies demonstrated that highly autoreactive iNKT cells carry innate-type TCRs, recognize CD1d molecules in a ligand-independent manner and evoke severe systemic autoimmune responses within a few hours upon adoptive transfer. In contrast, iNKT cells with intermediate to low autoreactivity express adaptive-type TCRs, which selectively recognize self-ligands and induce tissue-specific moderate autoimmune responses in vivo. High throughput CDR3β sequencing revealed that the frequency of highly autoreactive human iNKT cells positive for all 3 motifs was significantly reduced among iNKT cells in the periphery compared to the thymus. We conclude that intermediately and lowly autoreactive iNKT cells which prevail in the human periphery are adaptive lymphocytes and that the sole adaptive CDR3β sequences regulate the antigen specificity and reactivity of iNKT cells to avoid harmful autoimmune responses.
IL-33/ST2 promotes tissue mastocytosis under conditions of inflammation (CCR3P.219)
Authors: Wang Jun-Xia , Kaieda Shinjiro , Nigrovic Peter 
Published: January 2023

Abstract
Mast cells (MC) are potent innate immune cells that accumulate in chronically inflamed tissues, where they have been implicated in multiple allergic and inflammatory diseases. MC express the IL-33 receptor ST2, and this new member of the IL-1 cytokine family is recognized as an important pathway both for direct MC activation and for priming MC to respond to other pro-inflammatory signals. However, IL-33 has no known role in tissue mastocytosis. Using primary MC cultured from human skin, we found that IL-33 did not alter MC proliferation but instead protected MC from apoptosis, principally through upregulation of the anti-apoptotic molecule BCLXL. Murine bone marrow-derived MC demonstrated a similar response to IL-33, dependent entirely upon ST2, allowing us to test the role of this biology in vivo. Mice lacking ST2 exhibited fewer peritoneal MC as well as reduced accumulation of MC in inflamed arthritic joints and helminth-infected intestine. Upon engraftment into the peritoneum, WT MC exhibited a clear, IL-33-mediated survival advantage over ST2-deficient MC, in particular after thioglycollate-induced peritonitis. Together, these data reveal a novel, cell-intrinsic role for the IL-33/ST2 axis in tissue mastocytosis, especially under conditions of local inflammation.
Significance of CXCR6-CXCL16 in prostate cancer progression and therapeutic efficacy (CCR3P.222)
Authors: Gupta Pranav , Sharma Praveen , Grizzle William , Lillard James , Singh Shailesh 
Published: January 2023

Abstract
Despite recent advances prostate cancer (PCa) is still a leading cause of cancer related deaths among men, primarily because etiopathogenesis of PCa is not well defined. Chemokines and their corresponding receptors have been shown to play major role in PCa progression and poor therapeutic outcomes. The focus of this study is to determine the potential role of CXCR6 and its natural ligand CXCL16 in PCa pathogenesis and impact of this axis on docetaxel (DTX) efficacy. We investigated CXCR6/CXCL16 expression in clinical samples and prostate cell lines. CXCR6/CXCL16 expression was significantly higher in PCa samples compared to their respective controls, which showed positive correlation with tumor stage and grades. Similar to patient’s samples, PCa cell lines showed higher expression of CXCR6 and CXCL16 compared to normal prostatic epithelial cells. Furthermore, molecules involved in cell survival, matrix digestion and epithelial-mesenchymal transition were differentially modulated by CXCL16 treatment. Interestingly, expression of CXCR6/CXCL16 was elevated in PCa cells following DTX treatment compared to untreated cells. Taken together, better efficacy of DTX was observed when given with anti-CXCR6 is presumably due to inhibition of CXCR6-mediated survival signals. These findings suggest that CXCR6-CXCL16 axis play a crucial role in PCa pathogenesis and inhibition of this axis may have significant impact on disease progression and therapeutics outcomes.
HMGB1 is a key mediator of anemia in a murine model of sepsis survivors (CCR3P.224)
Authors: Valdes-Ferrer Sergio , Blanc Lionel , Chavan Sangeeta , Tracey Kevin 
Published: January 2023

Abstract
Anemia is omnipresent in patients with severe sepsis, and correlated to long term outcome. Its mechanism is incompletely understood. High mobility group box 1 (HMGB1) is persistently elevated in murine sepsis survivors. To study the role of HMGB1 in anemia of sepsis, we used a murine model of severe sepsis induced by cecal ligation and puncture (CLP). Anemia developed 3 days after onset of sepsis and persisted for no less than 28 days. The compensatory regenerative response was blunted, as demonstrated by reduced and erratic reticulocytosis. Splenomegaly was present in CLP survivors for as long as the follow up period. Microscopy revealed splenic architecture disruption, with expansion of the red pulp, characteristic of stress erythropoiesis. Analysis of terminal erythroid differentiation using flow cytometry demonstrated an increase in all erythroid progenitors, from proerythroblast to orthochromatic erythroblast, while mature erythroid cell populations were significantly reduced. An anti-HMGB1 (2G7 clone; 50ug/day x3 days I.P.) administered 9 days after after CLP significantly protected septic mice from anemia. In contrast, exogenous administration of HMGB1 (500ug/mouse/day x7d I.P.) to healthy mice was sufficient to induce a significant decrease in hemoglobin (p&gt;0.05) and splenomeglay. These results indicate HMGB1 is necessary and sufficient for developing anemia in murine sepsis, and suggest HMGB1 may be a novel therapeutic target of anemia in sepsis survivors.
BAFF induces IL-6 production by monocytes through a BAFF receptor and promotes IgG production by B cells in patients with primary Sjögren's syndrome. (CCR3P.210)
Authors: Yoshimoto Keiko , Ishioka Eriko , Nishikawa Ayumi , Suzuki Katsuya , Kameda Hideto , Abe Tohru , Takeuchi Tsutomu 
Published: January 2023

Abstract
Backgrounds and Objectives: We have found that the elevated expression of BAFF receptor (BR3) on monocytes is involved in activation of monocytes to promote IL-6 production in patients with primary Sjögren's syndrome (pSS). In addition, it is well known that pSS is often accompanied by hypergammaglobulinemia (HγG). In our present study, we investigated the possible involvement of monocytes in the development of HγG. Methods: Peripheral monocytes were cultured in vitro with or without peripheral B cells and stimulated with soluble BAFF (sBAFF). The amounts of IL-6 and IgG in the culture supernatants were measured by ELISA. FACS analysis of whole blood samples was employed to analyze the expression of BR3. Results: The serum level of IgG and the proportion of BR3-positive monocytes (BR3+/CD14+) were elevated in pSS patients as compared to those in normal individuals. Remarkably, the BR3+/CD14+ ratio was positively and significantly correlated with the serum IgG level. In addition, the ratio was positively and significantly correlated with the amount of IL-6 produced by pSS monocytes stimulated with sBAFF. Stimulation of co-culture of B cells and monocytes with sBAFF drastically enhanced IgG production by B cells. Conclusions: Our data collectively suggest that the abnormal expression of BR3 on monocytes is responsible for the overproduction of IgG by B cells in pSS patients.
Metabolic maturation, migration, and liver retention of CD4 single positive thymocytes and recent thymic emigrants (HEM4P.238)
Authors: Ge Qing , Xu Xi , Zhang Shusong 
Published: January 2023

Abstract
CD4 or CD8 single positive (SP) thymocytes, recent thymic emigrants (RTEs), and peripheral naïve T cells represent three consecutive cell populations along T cell maturation program. Except for negative selection in SP thymocytes, the importance and the regulation of this maturation is not well understood. In particular, little is known about the transition from mature SP to RTE and key events of this transition during and after thymic egress. We identified the phenotype of CD4+ pre-RTEs, a population of SPs that has acquired the thymic egress capability. A pro-oxidant shift and an increase of mitochondria content was found in pre-RTEs when compared to SPs with less functional maturity. The elevation of intracellular ROS was involved in promoting the functional maturation of these young T cells. After egress, autoreactive CD4+ pre-RTEs were retained and some were activated in the liver. Education from secondary lymphoid organs were not required for hepatic retention but TCR-MHC interaction in the liver was important for the activation of CD4+ liver RTEs. When transferred into RAG-/- mice, liver but not lymph node RTEs caused T cell infiltration and severe pulmonary inflammation, suggesting that the liver may be the first checkpoint in the periphery to filter and retain autoreactive CD4+ RTEs that escape from negative selection. Together, our results emphasize the complexity of SP-RTE maturation and the impact of various lymphoid tissue microenvironments on this maturation.
Cell types important for fostering normal thymic medullary epithelial cell development (HEM4P.242)
Authors: Jeon Hyein , Williams Joy , Hodes Richard 
Published: January 2023

Abstract
Tolerance in developing T cells is dependent on medullary thymic epithelial cells (mTEC) and mTEC development in turns requires signals from mature SP thymocytes. Several TNF family receptor-ligand pairs have been shown to be important players in the thymic crosstalk required to foster mTEC development including RANK-RANKL, LTαβ-LTβR and CD40L-CD40. In addition to confirming the important roles of TNF family members in thymic crosstalk, we have recently identified CD28-CD80/86 interactions as an additional pathway important for normal thymic medullary compartment development. CD80 and CD86, similar to LTβR, CD40 and RANK, are expressed by mTEC as well as by bone marrow (BM) derived cells in the thymus. To address the question of which cell type(s) need to express CD80/86, LTβR, CD40 and RANK in order to support mTEC development, we have generated and analyzed BM chimeras or conditional knockout mice in which either non-BM-derived cells (including thymic epithelium) or BM-derived cells express the molecule in question. For LTβR and RANK, we find that expression on radiation-resistant non-BM-derived cells, is required. Interestingly, however, we find that for CD80/86 and CD40, expression on either non-BM-derived or BM-derived cells is sufficient to promote mTEC development. Our findings suggest that models of thymic crosstalk must consider not only communication between TEC and thymocytes but with other bone marrow derived cell types as well.
Exploring the thymic niche requirements for regulatory T cell development (HEM4P.230)
Authors: Weist Brian , Robey Ellen 
Published: January 2023

Abstract
Development of regulatory T cells in the thymus is crucial for maintenance of tolerance to self. Here we demonstrate a new approach to systematically study the niche requirements for development of regulatory T cells in thymus. Using a thymic slice model, delivery of a model antigen leads to synchronized Treg development over the course of 72 hours. Antigen delivered by an exogenous antigen loaded dendritic cell population, or expressed as a transgene in medullary thymic epithelial cell populations, or a mixture of the two results in similar Treg numbers developing, and in all cases the number of developing regulatory T cells does not exceed 20% of antigen specific thymocytes. Addition of IL2 to thymic slices enhanced development, providing evidence that IL2 availability is a limiting factor for Treg development. We observed diminished Treg development when IL2-/- dendritic cells are used to present antigen, providing evidence that the APC population can be an important source of IL2 for developing Tregs. Overall, the thymic slice model provides an excellent system to dissect the niche requirements for the development of Tregs. Here we show that both antigen availability and IL2 exposure from a novel source can influence Treg development, but additional factors remain to be uncovered.