bluuebunny/crossref_metadata_2025_split huggingface dataset:
Trogocytosis and endocytosis of HLA-G from extravillous trophoblasts by decidual NK cells and its functional consequence (MUC2P.922)
Authors: Tilburgs Tamara , Evans James , Crespo Angela , Strominger Jack 
Published: January 2023

Abstract
The interaction of non-cytotoxic decidual NK cells (dNK) and extravillous trophoblasts (EVT) at the fetal-maternal interface has been studied. Confocal microscopy revealed that many dNK can interact with a single large EVT, leading to the formation of synapses. Filamentous projections from EVT enriched in HLA-G were shown to contact dNK and may represent the initial stage of synapse formation. A small fraction of dNK as isolated (~3%) contain surface HLA-G. However, HLA-G negative dNK had internalized HLA-G. Activation of dNK by culture in IL-15 resulted in disappearance of the internalized HLA-G in parallel with restoration of cytotoxicity. Surface HLA-G could be reacquired by incubation with EVT. However, under the conditions of the current experiments, re-acquisition of surface HLA-G by IL-15 activated dNK did not suppress their cytotoxicity. These data indicate a cycle in which HLA-G is acquired from EVT, endocytosed, degraded and, finally, reacquired from EVT. The HLA-G cycle is related to the loss and reacquisition of dNK cytotoxicity. The fetal-maternal interface offers an opportunity to study how evolution has met the challenge of generating a localized and specific tolerized immune compartment that does not reject an allogeneic fetus while maintaining the ability to respond to infections at this site.
The effect of sympathetic innervation on murine corneal epithelial wound healing (MUC3P.944)
Authors: Li Zhijie , Xue Yunxia , Smith Wayne 
Published: January 2023

Abstract
This study investigated the influence of murine corneal sympathetic nerves on epithelial wound healing. The wound model involved mechanical removal of a 2mm diameter area of the central corneal epithelial layer. We evaluated re-epithelialization, leukocyte extravasation, and platelet accumulation in the limbus and cornea at different time intervals after injury. The potential influence of sympathetic nerves in healing and inflammation was assessed by chemical sympathectomy, as well as topically applied adrenergic agonists and antagonists. Sympathectomy and the β2 receptor antagonist, timolol, each significantly accelerated the corneal epithelial healing process, including re-epithelialization and epithelial cell division. The agonist isoproterenol significantly inhibited re-epithelialization and epithelial cell division. Neutrophil migration into the wounded cornea was significantly inhibited by either sympathectomy or topical timolol, while isoproterenol restored neutrophil migration in the sympathectomized corneas. Platelet accumulation was inconsistently influenced by sympathectomy and timolol, but markedly inhibited by isoproterenol. These results are consistent with the interpretation that corneal sympathetic nerves can modulate corneal epithelial wound closure and platelet accumulation, and augment neutrophil influx.
Development of anti-CXCL13 monoclonal antibody for the treatment of autoimmune and immuno-oncological disorders (THER2P.950)
Authors: Klimatcheva Ekaterina , Mallow Crystal , Reilly Christine , Torno Sebold , Bussler Holm , Howell Alan , Scrivens Maria , Wang Wei , Balch Leslie , Murante Frank , Foster Cathrine , Kirk Renee , Giralico Susan , Leonard John , Paris Mark , Fisher Terrence , Evans Elizabeth , Smith Ernest , Zauderer Maurice 
Published: January 2023

Abstract
The chemokine CXCL13 is expressed in secondary lymphoid organs by follicular dendritic cells and macrophages and is also expressed by TH17 cells. It is the only known ligand for the CXCR5 receptor expressed on mature B cells, multiple subsets of T cells, macrophages and endothelial cells. Aberrant expression of CXCL13 within ectopic germinal centers has been linked to the development of autoimmune and cancer-promoting inflammatory disorders. Over-expression of CXCR5 receptor in certain cancers has been reported to induce CXCL13-dependent cell proliferation and metastasis. We developed a human IgG1 monoclonal antibody that specifically binds to human, rodent and primate CXCL13 and is capable of neutralizing CXCL13 function of these various species in in vitro functional assays. For preclinical in vivo studies we have engineered a chimeric antibody that contains the same human heavy and light chain variable genes as the human antibody along with mouse constant regions. This antibody has demonstrated efficacy in murine models of autoimmunity (Experimental Autoimmune Encephalomyelitis, Collagen-Induced Arthritis) and immuno-oncology (MALT lymphoma). In adoptive transfer and immunization studies, treatment with this antibody reduced B cell trafficking to the spleen and lymph nodes and interfered with the formation and expansion of germinal centers. The human antibody is currently undergoing pre-clinical development to support a future FDA Investigational New Drug application.
Nicotine exerts protective effect on ulcerative colitis via miR-124/STAT3 pathway (THER2P.962)
Authors: Liu Xia 
Published: January 2023

Abstract
A large body of epidemiological studies showed that smoking has a protective effect in patients with ulcerative colitis (UC). However, the underlying mechanism remains unknown. We previously reported nicotine could inhibit inflammation via miR-124-STAT3 pathway. Whether this pathway is involved in the protective effect of nicotine, accounting for the beneficial effect of smoking on UC, should be further explored. We found nicotine treatment could hamper weigh loss, improve disease activity index, decrease HE score, and increase miR-124 expression in the colon tissue of UC mice. Colon biopsies from patients with UC also showed higher miR-124 expression when compared with normal colons. Furthermore, miR-124 knockdown could markedly impair the beneficial role of nicotine in UC mice, and attenuat the inhibitory role of nicotine in the expression of STAT3 /p-STAT3 in colon tissues. In vitro, nicotine elevated miR-124 expression in colon epithelial cell HT-29, and miR-124 agomir or antagomir could decrease or increase STAT3/p-STAT3 expression and TNF-a production in these cells. Our data provide the first evidence that miR-124-STAT3 pathway is essential for the nicotine therapeutic effect on UC, and miR-124 is a potential therapeutic target for the treatment of UC.
Immune responses to chimeric human-FIX-mouse-Fc fusion protein in a hemophilia B mouse model (THER2P.963)
Authors: Golding Basil , Jiang Haiyan , Sauna Zuben , Levin Ditza 
Published: January 2023

Abstract
The fusion of a protein with the Fc-region of an antibody is used to increase the circulating half-life of therapeutic proteins. Using a mouse hemophilia B (HB) model we compared the immune responses to infusions of recombinant human FIX (rFIX) and human FIX-mouse Fc (hFIX-mFc) fusion protein. HB mice received weekly IV injection of rFIX or hFIX-mFc for 5 weeks. Anti-FIX IgG, IgE and cytokines were measured. Splenic cells were used to determine FIX-specific T and B cell functional and memory responses. Our results showed that, following IV treatment with rFIX or hFIX-mFc, there are no statistically significant differences in the titers of anti-FIX IgG Ab and numbers of long term memory B cells. However, a significantly larger proportion of mice treated with rFIX developed elevated titers of total IgE which correlated with a Th2 bias following rFIX. In contrast, hFIX-mFc induced a Th1-like response and no detectable IgE. Our study suggests, that in mice, hFIX-mFc fusion protein promotes a Th1-like response and is less likely than rFIX to induce an IgE response, which may lead to anaphylaxis, a rare severe adverse event associated with the treatment of HB patients.
IL-10 is critical in the regulatory function of CD1dhiCD5+ B cells in Experimental Autoimmune Myasthenia Gravis (THER2P.964)
Authors: Sheng JianRong , Soliven Betty 
Published: January 2023

Abstract
Autoimmune Myasthenia Gravis (MG) is a T cell-dependent, B cell-mediated autoimmune disorder in which autoantibodies is produced targeting the skeletal muscle acetylcholine receptor (AChR). IL-10-competent subset within CD1dhiCD5+ B cells, also known as B10 cells, has been shown to regulate autoimmune diseases. In our previous study, adoptive transfer of CD1dhiCD5+ B cells expanded in vivo by GM-CSF prevented and suppressed Experimental Autoimmune Myasthenia Gravis (EAMG), which was associated with downregulation of mature dendritic cell marker and expansion of regulatory T cell resulting in the suppression of acetylcholine receptor (AChR)-specific T cell and B cell response. To further examine the role of IL-10 in the regulatory function of B10 cells in EAMG, we induced EAMG in both wild type and IL-10 knockout mice, and also performed the adoptive transfer IL-10 deficient CD1dhiCD5+ cells to EAMG mice. We found that IL-10 knockout mice showed more severe EAMG disease with more Th1 cytokine production and less Foxp3 expression compared with wild type mice. Adoptive transfer of CD1dhiCD5+ B cells from IL-10 knockout mice to EAMG mice could not prevent and suppress disease. Our observations further confirmed that IL-10 derived from B10 cells played a critical role in maintaining and restoring immune homeostasis in EAMG mice. Administration IL-10 competent CD1dhiCD5+ B cells may represent an effective strategy in the treatment of human myasthenia gravis.
Autophagy reduces the accumulation of misfolded HLA-B27 in a rat model of spondyloarthritis (THER2P.951)
Authors: Navid Fatemeh , Layh-Schmitt Gerlinde , Colbert Robert 
Published: January 2023

Abstract
The MHC class I allele HLA-B27 is strongly associated with the development of spondyloarthritis (SpA) in humans, and when expressed in rats causes a SpA-like inflammatory disease. HLA-B27 has the tendency to misfold, and when up regulated by IFNγ and TNFα can cause ER stress leading to an unfolded protein response (UPR), which promotes pro-inflammatory cytokine production and may be critical for disease development. We examined whether alteration of autophagy affects HLA-B27 misfolding and UPR activation. Bone marrow-derived macrophages were obtained from HLA-B27-transgenic (TG), HLA-B7-TG and Lewis control (WT) rats and autophagy was induced with rapamycin. Western blot (WB) and immunofluorescence analysis of LC3B-II expression revealed a normal autophagic response in HLA-B27 expressing cells. Activation of autophagy with rapamycin reduced the accumulation of misfolded HLA-B27 induced by TNFα and IFNγ by 50%, while properly folded HLA-B27 heavy chains were minimally affected. Inhibition of autophagic flux with bafilomycin resulted in an exacerbation of the UPR upon HLA-B27 upregulation as measured by XBP1 splicing and expression of UPR target genes. These results demonstrate for the first time that induction of autophagy can reduce the accumulation of misfolded HLA-B27, and blocking autophagic flux exacerbates the UPR in HLA-B27 expressing cells. Alteration of the autophagic pathway may be a promising therapeutic tool in the treatement of SpA.
Characterization of the function and co-expression patterns of human FOXP3 isoforms and their role in IPEX syndrome development (THER2P.952)
Authors: Darrasse-Jeze Guillaume 
Published: January 2023

Abstract
The transcription factor FOXP3 plays a critical role in the development and function of regulatory T cells (Tregs) that maintain self-tolerance. Contrary to mice, humans possess three isoforms of FOXP3 with unknown function, and the expression of these isoforms is not restricted to Tregs. We determined the levels of expression and distribution of the isoforms of FOXP3 as well as other transcription factors in normal human T-cell subsets and in T cell subsets of patients with autoimmunity at the single-cell level by multiplex RT-qPCR and flow cytometry. We observed that the expression of these isoforms is not restricted to Tregs, but can also be detected in naïve and activated/memory helper (Th) and in activated/memory CD8+ T cells. Distinct patterns of co-expression of the FOXP3 isoforms are modulated in response to inflammatory or autoimmune stimuli. Moreover, we observed that normal FOXP3 isoforms could induce bona fide regulatory function in Th cells. Our work supports the view that lack of FOXP3 expression in the effector T-cell populations in IPEX patients may contribute to the pathogenesis of the disease, and our preliminary data suggest that FOXP3 deficiency can be corrected by gene transfer of the appropriate FOXP3 isoforms into conventional T-cells, which has important implication for the treatment of IPEX patients.
The potent, selective RORγt blocker, VTP-43742, suppresses Th17 production in vivo and provides greater benefit than IL-17 blockade in the EAE model of autoimmunity (THER3P.968)
Authors: Zhao Yi , Meng Shi , Noto Paul , Bukhtiyarov Yuri , Stadanlick Jason , Zhuang Linghang , Dillard Lawrence , Claremon David , Gregg Richard , McGeehan Gerard , Lala Deepak 
Published: January 2023

Abstract
Antibodies targeting IL-23 or IL-17 are highly effective in the treatment of psoriasis and psoriatic arthritis, validating the RORγt /Th17 pathway in human disease. VTP-43742 is a potent, selective and orally active inhibitor of RORgt that is being pursued as a treatment of autoimmune disorders. Herein, we present data showing the effects of VTP-43742 in two separate mouse studies using the MOG35-55/CFA immunized EAE model. In a subchronic study, prophylactic dosing of VTP-43742 for 11 days gave a dose-dependent suppression of both CD4+IL-17+IFNγ- and CD4+IL-17+IFNγ+ T cells in spleen and in draining lymph nodes. Splenocytes from these mice restimulated in vitro with the MOG35-55 peptide showed specific suppression of Th17 derived cytokines. In a chronic disease study, the maximal pharmacological effect of VTP-43742 was compared to maximal IL-17 blockade by mouse monoclonal antibody MM17F3 (eBioscience) in a prophylactic setting. High dose VTP-43742 completely suppressed the EAE clinical score, significantly beyond the reduction seen with MM17F3 (p&lt;0.001). Analysis of spinal cord RNA showed significant suppression of Th17-associated markers by VTP-43742 with no significant reduction by MM17F3. Additionally, spinal cord histology showed qualitatively greater suppression of demyelination and inflammatory cell infiltration with VTP-43742 treatment versus MM17F3, suggesting that RORγt blockade may provide treatment benefit beyond anti-IL-17 targeted strategies in autoimmunity.
Metformin ameliorates inflammatory bowel disease by suppression of STAT3 signaling pathway and regulation of the Th17/Treg balance (THER3P.1001)
Authors: Lee Seon-Young , Lee Seung Hoon , Kim Eun-Kyung , Kim Jae-Kyung , Shin Dong-Yun , Cho Mi-La 
Published: January 2023

Abstract
Metformin is an effective pharmacological treatment for type 2 diabetes. The current study attempts to determine whether metformin reduces inflammation regulating p-STAT3 expression and Th17 cell proliferation in an experimental inflammatory bowel disease (IBD) mice model. IBD mice were intrarectally injected with either metformin once a day for 16 days. Histological analyses of the colons were performed via immunohistochemistry. AMPK, mTOR, p-STAT3 and p-STAT5 of the spleen and lymph node tissue were detected by confocal microscopy staining. Gene expression was calculated by real-time PCR and protein levels were measured using western blot enzyme-linked immunosorbent assay (ELISA). MTT assay was used to evaluate human HT-29 cell proliferation. Metformin reduced DAI score and inhibited weight loss. Metformin also decreased the colonic histological score and inflammatory mediators with increased colon length. Expression of IL-17, p-STAT3 and p-mTOR was inhibited with metformin treatment. On the contrary, metformin treatment increased the expression of p-AMPK and Foxp3. Additionally, inflammatory cytokines expression was reduced dose dependently in inflamed human HT-29 cells cultured in various concentrations of metformin treatment. Metformin attenuates the severity of IBD and declines inflammation through the inhibition of p-STAT3 and IL-17. These results support targeting therapeutic substances for IBD and to increase the understanding of IBD.
Immune cell responses to combined treatment of murine melanoma with ALT-803 and Sunitinib in normal and chronic alcohol consuming mice (TUM2P.1032)
Authors: Little Alexander , Gaither Kari , McBride Alisha , Castillo Savanna , Brar Kiran , Platt Amity , Zhu Zhaohui , Zhang Faya , Luong Dung , Zhang Hui , Meadows Gary 
Published: January 2023

Abstract
The immune enhancing IL-15 super agonist ALT-803 and Sunitinib, a tyrosine kinase inhibitor that inhibits immunosuppressive myeloid derived suppressor cells (MDSC) and T regulatory (Treg) cells, were examined for their single and combined effects on immune cell phenotype in normal and chronic alcohol consuming (CAC) mice inoculated with s.c. B16BL6 melanoma. ALT-803 alone and in combination with Sunitinib initially increased CD8+CD44hi and CD8+CD44hiCD62Llo memory phenotype T cells in the spleen and PBL. IFN-γ producing CD8+CD44hi T cells increased in the spleen and the increase was more robust in CAC versus normal mice. These responses decayed during the later stages of tumor growth, and declined more rapidly in the CAC mice. Sunitinib alone did not affect CD8+ T cells or their IFN-γ production. Sunitinib alone and in combination with ALT-803 prevented the increase in CD11b+Gr-1int MDSC in the spleen associated with no treatment or ALT-803 treatment in normal mice; however, the effect was less pronounced in CAC mice where MDSC did not increase in the untreated group over time. MDSC in the PBL increased over time in untreated and ALT-803-treated normal mice. Sunitinib prevented this increase until the late stages of tumor progression. Sunitinib alone and in combination suppressed CD4+CD25+FoxP3+ Treg cells in the spleen during tumor progression.
Oral vaccine by tumor cell-derived microparticles leads to effective anti-tumor immune responses (TUM2P.1041)
Authors: Huang Bo 
Published: January 2023

Abstract
In response to stimuli, tumor cells release size of 100-1,000 nm vesicles, called microparticles (MP) to the extracellular space. Our previous studies have demonstrated that tumor cell-derived microparticles (T-MP) can act as cell-free tumor vaccines. T-MPs present both tumor antigens and immunostimulatory signals to dendritic cells, leading to tumor-specific T cell responses. Here, we further examine whether the T-MPs are capable of inducing anti-tumor immune responses when administrated via oral route. We find that treatment with oral B16 melanoma-derived MPs effectively inhibits tumor growth and improves survival of B16 melanoma-bearing mice, compared with tumor lysate or whole tumor cell control group. However, these MPs do not induce the anti-tumor effect in nude mice. In addition, depletion of CD4 or CD8 T cells impairs T-MP-mediated anti-tumor immunity in immune-competent mice. Importantly, mice orally provided with MPs derived from OVA-B16 cells display significantly increased frequencies of OVA-positive CD8+ T cells in the blood and spleen. Also, the levels of IFN-γ are enhanced in mice immunized with OVA-B16-MPs. Interestingly, we find that epithelial cells transport T-MPs to dendritic cells, resulting in T cell responses. We use DC/EC coculture system and demonstrate that the transepithelial transport of T-MPs is mediated by the epithelial transcytosis. Collectively, our data suggest that oral delivery T-MPs elicit an effective anti-tumor immune responses.
A quantitative approach to T cell killing (TUM2P.1043)
Authors: Whitlock Ben , Altan-Bonnet Gregoire 
Published: January 2023

Abstract
Previous research has identified the pathways and proteins cytotoxic T lymphocytes (CTLs) use to kill antigen presenting cells (APCs); however, it is currently impossible to predict how many T cells are needed to kill a specific number of APCs in vivo. Current methods for examining killing rely on bulk measurements and effector to target ratios of 30:1, which are thought to be unphysiological. We have developed a single-cell killing assay that occurs within a 50x50μm well, which we image with time-lapse fluorescent microscopy. This has allowed us to make new observations that would have been impossible to discern using conventional bulk killing assays. Using this approach, we observe that T cells have a greater affinity for living cells than dead cells. We hypothesize that this phenomenon could be a way for T cells to avoid dead cells in vivo and are performing additional experiments to examine this.
Unmasking cancer to innate sensing by anti-CD47 antibody triggers adaptive immune-mediated tumor destruction (TUM3P.1045)
Authors: Xu Meng 
Published: January 2023

Abstract
Using xenograft and in vitro models, anti-CD47 antibody is thought to enhance "eat me" signaling to kill tumor cells by macrophage-mediated phagocytosis. However, the potential for adaptive immune response in anti-CD47 mediated tumor control has been ignored. Here we reveal a novel mechanism in which tumor regression after local anti-CD47 antibody treatment depends on cytotoxic T lymphocyte (CTLs) response. Mechanically, tumor recognition by DC following anti-CD47 enhances type I IFNs production and that subsequent IFNs signaling strengthens the cross-priming capability of DCs within the tumor microenvironment. Intriguing, anti-CD47 mediated antitumor immunity requires adaptor protein STING for the innate sensing of tumor cells. Additionally, we delineate a clinically relevant combinational strategy of chemotherapeutic drugs and anti-CD47 antibody to enhance the reduction of tumor burden without abrogating anti-CD47 initiated immunity.
Selective inhibition of the T cell p38 alternative activation pathway and pancreatic cancer (TUM3P.1046)
Authors: Alam Muhammad , Gaida Matthias , Bergmann Frank , Lasitschka Felix , Giese Thomas , Giese Nathalia , Hackert Thilo , Hinz Ulf , Hossain S. , Kozlov Serguei , Ashwell Jonathan 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive neoplasm with a five year survival rate of ~5%, that is characterized by a marked fibro-inflammatory microenvironment, the presence of which can promote cancer induction and growth. Therefore, selective manipulation of local cytokines is an attractive if unrealized therapeutic approach. T cells possess a unique mechanism for activation of p38 MAPK downstream of T cell receptor (TCR) engagement in which it is phosphorylated on Tyr-323 (pY323), which is required for pro-inflammatory cytokine production. Analysis of 192 human pancreatic carcinomas revealed that having a high percentage of infiltrating pY323+ T cells was accompanied by large numbers of TNFa and IL-17-producing CD4+ TIL and very aggressive disease. Cancer progression was inhibited in two different murine pancreatic tumors models by genetic ablation of the alternative p38 activation pathway. Strikingly, a plasma membrane-permeable peptide derived from Gadd45α, the naturally-occurring inhibitor of p38 pY323+, reduced infiltrating CD4+ T cell production of TNFa, IL-17A, IL-10, and induction of secondary cytokines, slowed the growth of implanted tumor cells and inhibited progression of spontaneous K-ras-driven adenocarcinoma. Thus, TCR-mediated alternative p38 activation in tumor-infiltrating CD4+ T cells, which promotes the production of pro-tumorigenic inflammatory factors, can be targeted for therapeutic benefit.