bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhancing adoptively transferred T cells by stimulating with dendritic cell stimulation: exploring novel therapies (P4405)
Authors: Murakami Kiichi , Kawamura Kim , Chan Olivia , Tran Michael , Elford Alisha , Nguyen Linh , Ohashi Pamela 
Published: January 2023

Abstract
Adoptive Cell Transfer (ACT) of tumor infiltrating lymphocytes (TILs) is a promising strategy for cancer immunotherapy. The current therapy relies on non-myeloablative lymphodepletion and high dose IL-2 therapy, which has serious side effects for patients. Optimization of the current ACT therapy, using animal models, may help reduce or eliminate these additional treatments and enhance the response rate. ACT has been evaluated in the autoimmune model RIP-GP mice, which express the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) on the β-islet cells of the pancreas. Our findings showed that LCMV memory T cells stimulated with matured dendritic cells (DCs) and pulsed with peptides derived from LCMV, successfully induced diabetes. These stimulated T cells expanded well in vivo and showed strong killing activity after transfer. However, unstimulated memory tissue specific T cells rarely expanded after transfer and showed limited cytolytic activity. To evaluate this method in the tumor model, RIP-Tag2/GP double transgenic mice were used, which express the large T antigen under the control of RIP and spontaneously develop insulinomas. Islet specific T cells cocultured with DCs and LCMV-GP peptides displayed strong anti-tumor activity after transfer in vivo. Surprisingly, mice bearing insulinomas survived longer after ACT than mice treated with LCMV vaccination. Therefore, stimulating TILs before infusion has high potential for enhancing ACT therapy.
Characterization of a human CD27 agonistic monoclonal antibody for use as an agent in glioblastoma therapy (P4411)
Authors: Riccione Katherine , Sanchez-Perez Luis , Flores Catherine , Sampson John 
Published: January 2023

Abstract
CD27 is a costimulatory molecule constitutively expressed on naïve T-cells, B-cells, and subsets of NK cells; signaling through CD27 has been shown to promote Th1 polarization, clonal expansion, enhanced survival of effector and memory cells, upregulation of effector molecules, and enhanced cytolytic activity. We are characterizing the use of an agonistic anti-CD27 antibody, CDX1127, as a treatment in a preclinical glioblastoma model in human CD27 transgenic mice. We have shown that in vitro stimulation of isolated T-cells from hCD27+/- mice with CDX1127, in the context of TCR signaling, leads to increased proliferation, decreased apoptosis, and increased secretion of IFN-γ and TNF-α, corroborating its agonistic activity. Furthermore, we have analyzed the efficacy of CDX1127 as a monotherapy in a mouse glioblastoma model as well as an adjuvant in a total tumor RNA dendritic cell vaccine.
A VCG-based chlamydial vaccine engenders broad-based protective immunity (P4391)
Authors: Eko Francis , He Qing , Black Carolyn , Igietseme Joseph 
Published: January 2023

Abstract
Genital chlamydial infections pose a considerable public health challenge worldwide and a prophylactic vaccine is required for control. Since immunity against natural infection is serovar specific, an efficacious chlamydial vaccine should induce broad-based immunity against the different chlamydial serovars. Intramuscular (IM) immunization with recombinant Vibrio cholerae ghosts (rVCG) expressing chlamydial antigens has shown promise in development of immunity against genital chlamydial infection. We evaluated the ability of a VCG-based multisubunit vaccine expressing the porin B (PorB) and polymorphic membrane protein D (PmpD) proteins of C. trachomatis to induce cross-protective immunity in a mouse model of genital chlamydial infection. IM immunization with the vaccine candidate elicited high levels of antigen-specific genital mucosal and systemic Th1 cell-mediated and humoral immune responses against heterologous serovars and strains, including serovars E-H and L. Also, the multisubunit vaccine conferred significant cross protection that was superior to that provided by VCG expressing the single antigens against the heterologous mouse strain, C. muridarum. These results demonstrate the ability of the serovar D-derived multisubunit vaccine to induce cross protective chlamydial immunity using the VCG platform.
Altered distribution patterns of TLR5 and TLR7 in the central airways of severe asthmatics (P4427)
Authors: Shikhagaie Medya , Bergqvist Anders , Andersson Cecilia , Mori Michiko , Krohn Kortekaas , Dahl Ronald , Hoffman Hans-Jurgen , Bjermer Leif , Erjefält Jonas 
Published: January 2023

Abstract
Toll-like receptors are a major class of pattern and damage recognition receptors controlling innate immune responses. TLR5 recognizes bacterial flagellin and TLR7 identifies single stranded viral RNA. The present study characterizes TLR5 and TLR7 expression pattern in central airways of asthmatics. Methods: Bronchial biopsies were obtained from healthy controls, uncontrolled asthmatics on inhaled glucocorticoids, and severe asthmatics on oral glucocorticoids. Subjects were classified according to GINA guidelines and asthma control score. Tissue sections were processed for immunohistochemical identification of TLR positive cells. Results: TLR5 immunoreactivity was identified in the airway epithelium, glands, smooth muscle, alveolar macrophages and plasma cells. In the airway epithelium, TLR5 had mainly a basolateral distribution. In severe asthmatics the overall and epithelial TLR5 immunoreactivity were significantly decreased. TLR7 displayed a distinct peri-nuclear expression. Quantification of the epithelial TLR7 immunoreactivity showed a significantly decreased expression in severe asthmatics compared to healthy controls and uncontrolled asthma. TLR7 expression was also detected in nerve cells, NK cells and mast cells. Conclusions: Our data suggest that patients with severe asthma have decreased expression of TLR5 and TLR7 compared to healthy controls and uncontrolled asthmatics. This altered expression may reflect a dysfunctional immune-regulation in severe asthmatics.
Hepcidin protects sepsis through regulating iron homeostasis to inhibit immunocyte apoptosis in mice (P4435)
Authors: Zeng CongLi , Zhang Kai , Chen QiXing , Fang XiangMing 
Published: January 2023

Abstract
Background: Hepcidin is a key regulator of iron metabolism and may have a critical role in modulating LPS-induced acute inflammatory responses. The role of hepcidin in cecal ligation and puncture (CLP) induced sepsis model was not clear. Methods: Adenovirus-mediated shRNA to inhibit the expression of hepcidin and a relative negative control were constructed and were administrated to mice through hydrodynamic injection via tail vein. Thirteen days later, the mice were subjected to CLP. Bacterial colony counts in blood and organs, reactive oxygen species (ROS) production in liver and immunocyte apoptosis in spleen were analyzed. Survival rate was estimated after 7 days. Results: The survival rate of hepcidin-knock down (KD) mice in CLP-induced sepsis was significantly decreased. Hepcidin-KD mice exposed to CLP showed more severe lung injury and increased inflammation cytokines. The serum iron concentration in hepcidin-KD mice after CLP was significantly higher, which resulted in more ROS production in liver and increased bacterial counts in blood and organs. In the spleen, there were significantly more apoptotic immunocytes in hepcidin-KD mice than those in controls. However, no significant differences in SOCS3 and phosphorylation of STAT3 level between the two groups were observed. Conclusions: Hepcidin may have a protective role in sepsis through regulating iron balance to inhibit bacterial infection and oxidative stress, which then protected immunocyte from apoptosis.
Monitoring epithelial cell responses to an IBD-associated bacterial pathotype using quantitative proteomics (P1352)
Authors: Lijek Rebeccah , Chan Eric , Fleming Mark , Starnbach Michael 
Published: January 2023

Abstract
Many lines of evidence suggest that the initiation of inflammatory bowel disease (IBD) involves abnormal host-microbial interactions at the intestinal epithelium. One microorganism that is commonly associated with the intestinal mucosa of IBD patients but not from healthy subjects is called adherent invasive E. coli (AIEC). AIEC are a new pathotype defined by their ability to adhere to and invade epithelial cells in the absence of known virulence factors associated with other pathogenic E. coli. Previous work has demonstrated that AIEC infection of epithelial cells induces cell-intrinsic responses such as the activation of autophagy markers, reduction of barrier function and secretion of IL-8. However, a comprehensive analysis of the cellular response to AIEC has not been performed. Here, we take a global approach to define how protein levels are altered in epithelial cells exposed to AIEC using quantitative mass spectrometry. This experimental strategy allows us to determine how the endogenous abundance of known and novel cellular proteins is altered by AIEC infection. Our preliminary results reveal specific differences between the epithelial cell response to adherent invasive E. coli compared to commensal E. coli not associated with IBD. Ongoing work examines whether any of the changes in protein abundance observed in response to AIEC infection are necessary for AIEC attachment, invasion, or intracellular growth.
The role of microRNA in T Helper cell plasticity (P1307)
Authors: Siede Julia , Baumann Claudia , Haftmann Claudia , Mashreghi Mir-Farzin , Löhning Max 
Published: January 2023

Abstract
Upon antigen encounter naive CD4+ T helper (Th) cells differentiate into distinct T helper cell subsets characterized by specific cytokine and transcription factor expression. It is currently debated whether these Th subsets have a fixed or flexible phenotype. Previously we showed that in the course of viral infection Th2 cells can be reprogrammed into a hybrid Th2+1 phenotype co-expressing Th1 and Th2 key transcription factors and cytokines. However, the T cell intrinsic control of subset-specific gene expression is only partially understood. Studies have indicated that post-transcriptional regulation via microRNA contributes to several aspects of T cell gene expression. Nevertheless, only few relevant miRNAs have been described so far. The aim of this project is to identify miRNA specifically expressed in novel hybrid T cell phenotypes as well as to analyze their impact on T cell plasticity and function. We used microarray analysis to identify 22 differentially expressed miRNAs in hybrid Th2+1 cells compared to both Th1 and Th2 cells. Within this subset 8 miRNAs were up and 11 miRNAs were down regulated. We found that type I IFNs are potent inducers of some of these miRNAs. Additionally, we observed an inverse correlation in expression of some miRNAs with their predicted targets that are known for their role in Th cell activation and differentiation. Overall, these miRNAs could prove to be important players in the induction and maintenance of Th2+1 cells.
Regulation of intrathymic γδ T effector subset differentiation by high mobility group box transcription factors (P1342)
Authors: Cho Ok Hyun , Kang Joonsoo 
Published: January 2023

Abstract
Innate-like γδ T effector subsets are exported from the thymus as “memory-like” cells prewired for rapid, specialized function. Previously, we showed that emergent γδ cell subsets distinguished by TCRγ and TCRδ usage in the fetal and adult thymus possess distinct global transcriptomes. In particular, the subset-specific combinatorial expression of High Mobility Group box transcription factors (HMG TFs) was shown to be a primary determinant of γδ effector differentiation. We showed that a quartet of HMG TFs, SOX4, SOX13, TCF1 and LEF1, are critical for the development and function of IL-17 producing γδ T effectors (Tγδ17). The TFs are found co-localized near the transcriptional regulatory sites of Rorc, Blk, Il17a and Gata3 in a subset-specific manner, suggesting that they directly regulate the expression of key proteins that control effector cytokine production. We also observed a positive correlation between active epigenetic modifications and subset-specific gene networks governing effector function, suggesting that the initial programming of γδ effector diversification involves selective chromatin modifications. While the detailed mechanism of HMG TFs cooperativity and counter-regulations is not fully understood, a key feature is predicted to be context-dependent interactions between SOX13 and TCF1 that result in diversified target gene regulation.
Acute-phase serum amyloid A causes Th1 differentiation and proliferation (P1325)
Authors: He Rong , Lui Asona , Hinton Michael 
Published: January 2023

Abstract
Host response to injury and infection is accompanied by a rapid rise of acute-phase proteins in the blood. Serum amyloid A (SAA) is a major acute-phase protein, and has been used as a marker for inflammatory diseases. However, its precise role in inflammation has not been defined. In our previous studies, we found that SAA can induce IL-8 and TNF-a secretion from peripheral blood neutrophils, and can be an endogenous ligand that induces the expression of interleukin-23 (IL-23), a hetero-dimeric cytokine that promotes autoimmune inflammation. SAA is found in very high levels in the arthritic joints of patients, along with a high concentration of certain subsets of T lymphocytes, notably helper T cell type 1 (Th1). In this study, we discovered that SAA can also stimulate CD4+ T cells to produce the cytokine interferon gamma (IFNg) and interleukin-2 (IL-2), and induce Th1 cell differentiation and proliferation. Our data showed that SAA-induced IL-2 production and cell proliferation can be blocked by the inhibitor of MAP kinase p38, and the phosphorylation of p38 indeed can be increased after SAA stimulation. These results suggest that SAA, which is found in arthritic joints, may play a role to induce Th1 cell production, and is perhaps a missing link in our understanding of Rheumatoid Arthritis.
Hypoxia inducible factor-1 alpha links reduced oxygen availability to the generation of anti-inflammatory IL-10 expressing T cells (P1328)
Authors: Clambey Eric , Westrich Joseph , Eltzschig Holger 
Published: January 2023

Abstract
Hypoxia, the state of limited oxygen availability, is a frequent metabolic constraint in physiologic and pathologic states. The impact of hypoxia and hypoxic signaling on T cell differentiation remains incompletely defined, especially regarding inhibitory pathways that may be induced to limit hypoxia-associated tissue damage. Here we identified CD4 and CD8 T cells expressing IL-10 in the context of inflammatory hypoxia in vivo. To test the consequence of limited oxygen availability on the generation of IL-10 expressing T cells, we analyzed T cell differentiation under hypoxic conditions. Hypoxic culture significantly induced IL-10 expression in both CD4 and CD8 T cells. Hypoxia further increased IL-10 expression across diverse differentiation conditions, including conditions known to induce IL-10 expression. The hypoxic enhancement of IL-10 producing T cells was independent of TGF-β and IL-27, but required signals from multiple cell types, as well as T cell-intrinsic expression of the transcription factor hypoxia inducible factor (HIF)-1α. HIF-1α was further required for IL-10 expression in regulatory T cells even in normoxia, consistent with impaired suppressive function of HIF-1α deficient Tregs in vitro and in vivo. In total, these data demonstrate that reduced oxygen availability, interpreted through the HIF-1α transcription factor, is a potent physiologic cue that can drive T cell differentiation towards IL-10 production, thereby limiting immune-mediated pathology.
SNAP-23-dependent surface trafficking of NOX2 regulates ROS-dependent surface up-regulation of BLT1 during exocytotic degranulation induced by Trichomonas vaginalis-derived LTB4 (P3351)
Authors: Shin Myeong Heon , Min Arim 
Published: January 2023

Abstract
Trichomonas vaginalis is a sexually transmitted parasite that causes vaginitis in women. T. vaginalis itself secretes lipid mediator LTB4. Mast cells are important effector cells for tissue inflammation during infection with parasites. In this study, we investigated signaling mechanisms of mast cell degranulation induced by T. vaginalis secretory products (TvSP). Herein, we found that SNAP-23-dependent surface trafficking of NOX2 is required for BLT1-dependent exocytotic degranulation of mast cells induced by LTB4 in the TvSP. Stimulation with TvSP induced p47phox phosphorylation, ROS generation, surface up-regulation of CD63 that is a marker for exocytosis in human mast cell cells. TvSP-induced ROS generation and exocytosis were strongly inhibited by treatment with BLT1 siRNA or NOX2 siRNA. In addition, LTB4 in the TvSP was found to induce trafficking of NOX2, LTB4 receptor BLT1, and CD63 to the cell surface. Moreover, we found that t-SNARE SNAP23 siRNA almost completely inhibited TvSP-induced surface trafficking of NOX2 and ROS generation, which down-regulated surface upregulation of BLT1 and CD63 in HMC-1 cells. In conclusion, our results suggest that SNAP23-dependent surface trafficking of NOX2 are essential for ROS-dependent BLT1 upregulation during exocytotic degranulation in human mast cells induced by T. vaginalis-secreted LTB4. These results can help to explain the signaling talk mechanism between T. vaginalis and mast cells during tissue inflammatory responses.
Entamoeba histolytica induces cell death in HepG2 cells via the change of protein O-GlcNAcylation (P3355)
Authors: Lee Young Ah , Min Arim , Kim Kyeong Ah , Cho Jin Won , Shin Myeong Heon 
Published: January 2023

Abstract
Entamoeba histolytica is an enteric tissue-invasion protozoan parasite that causes amoebic colitis and liver abscess in humans. E. histolytica can induce host cell apoptosis to evade host immune responses. Protein O-GlcNAcyation has been thought to contribute to the various cellular signal processes including apoptosis and proliferation. O-GlcNAc addition and removal are regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), respectively. However, it is unknown whether the alteration of O-GlcNAcylated protein level in host cells can affect E. histolytican-induced cell death. In this study, co-incubation of HepG2 cells with virulent amoeba HM-1 strain remarkably increased DNA fragmentation and LDH release compared to cells incubated with medium alone. In addition, Entamoeba HM-1 induced activation of caspases-3 and calpain in HepG2 cells. Amoebic adherence to the HepG2 cells showed the decrease of O-GlcNAcylated protein levels in HepG2 cells within 2 min, while non-virulent Rahman strain did not. Indeed, pretreatment of host cells with O-GlcNAcase inhibitors (PUGNAc or Thi-Met G) or OGA siRNA prevented the E. histolytica-induced protein deGlcNAcylation and cell death in HepG2 cells. These results suggest that decrease of protein O-GlcNAcylation is an important process required for hepatocyte cell death induced by adherence of E. histolytica.
Role of SERPINB3, SERPINB4, and their mouse homolog Serpinb3a in allergen-induced cutaneous inflammation. (P3347)
Authors: Sivaprasad Umasundari , Kinker Kayla , Gibson Aaron , Bass Stacey , Hershey Nicolas , Biagini Myers Jocelyn , Butsch Kovacic Melinda , Martin Lisa , Khurana Hershey Gurjit 
Published: January 2023

Abstract
The serine protease inhibitors SERPINB3 and SERPINB4 have been implicated in allergic disorders. We have recently demonstrated a role for the serine protease inhibitor Serpinb3a (the mouse homolog of SERPINB3 and SERPINB4) in a mouse model of asthma. Elevated levels of SERPINB3 and SERPINB4 are detected in the serum of patients with atopic dermatitis (AD) and they have been proposed as possible biomarkers for AD. Serine proteases are critical for epidermal barrier homeostasis and aberrant expression and/or activity of serine proteases have been associated with AD in human studies. We also observe a significant induction in the expression of Serpinb3a in a mouse model of cutaneous inflammation that has many similarities to AD. However the role of Serpinb3a in the skin is unknown. Using our mouse model of skin inflammation, we observed attenuation in trans-epidermal water loss, skin inflammation scores, decreased levels of total IgE, and pro-inflammatory markers like S100A8 in mice lacking Serpinb3a following exposure to Aspergillus fumigatus extract. In humans, we have identified a single nucleotide polymorphism (SNP) in the locus that encodes SERPINB3 and SERPINB4 that is associated with trans-epidermal water loss (p-val=0.031). In conclusion, we demonstrate that SERPINB3 and SERPINB4 contribute to cutaneous inflammation and work is ongoing to elucidate the role of this protein in regulating the skin barrier.
Anti-tumor activity of obinutuzumab and rituximab in a follicular lymphoma 3D model (P3287)
Authors: Bezombes Christine , Decaup Emilie , Jean Christine , Laurent Camille , Gravelle Pauline , Laurent Guy , Klein Christian , Varoqueaux Nathalie , Fournié Jean-Jacques 
Published: January 2023

Abstract
Follicular lymphomas (FL) account for 35-40% of all adult lymphomas. Treatment typically involves chemotherapy combined with the anti-CD20 monoclonal antibody rituximab (RTX). The development of type II anti-CD20 MAb obinutuzumab (GA101) aims to further improve treatment. Here we show using FL cells that RTX and GA101 displayed similar activity on RL cells cultured in 2D. As 2D culture cannot mimic tumor spatial organization, conventional 2D models may not reflect the effects of antibodies as they occur in vivo. Thus, we created a NHL 3D culture system, termed multicellular aggregates of lymphoma cells (MALC), and compared RTX and GA101 activity. Our results show that both antibodies display greater activity towards FL cells in 3D culture compared to 2D culture. Moreover, we observed that in the MALC model GA101 was more effective than RTX in inhibiting tumor growth through induction of (lysosomal) cell death and senescence; and in inhibiting intracellular signaling pathways such as mTOR, Akt, PLCgamma and Syk. Altogether, our study demonstrates that spatial organization strongly influences response to antibody treatment supporting the use of 3D models for the testing of therapeutic agents in NHL.
Identification of sulfatide reactive type II NKT cells using CD1d dimers (P3291)
Authors: Parish Stanley , Izhak Liat , Xia Zheng , Suzuki Motoshi , Terabe Masaki , Berzofsky Jay 
Published: January 2023

Abstract
Natural killer T cells (NKT) are a unique subset of lymphocytes that bridge the gap between innate and adaptive immunity by recognizing lipid antigen presented by CD1d. While type I NKT cells have been extensively studied by using CD1d multimers loaded with α-galactosylceramide (α-GalCer), a type I specific antigen, studying type II NKT cells has lagged behind, since loading sulfatide, a type II-specific lipid antigen, to the tetramer has been difficult to accomplish reliably. In this study, we have developed sulfatide loaded CD1d dimers that accurately identify type II NKT cells by flow cytometry. Using saposin C, a lipid loading chaperone protein, and acidic conditions, as found in the natural loading environment, we could create sulfatide-loaded CD1d dimers to identify this subset. Sulfatide-loaded CD1d dimers specifically identified type II NKT cells in both WT and type I NKT deficient mice, and were not observed in NKT cell-deficient CD1d-/- mice. The specificity of the binding through TCRβ was confirmed by TCRβ blockade. Consistent with previous findings, in the liver, the sulfatide-reactive type II NKT cells were a distinct population that encompasses about 2% of liver lymphocytes. These data suggest that the method we developed can create sulfatide-loaded CD1d dimers to identify type II NKT cells, and has enabled us to identify other markers of this subset. This method can be used to load other lipids onto CD1d to identify other NKT subsets across multiple models.