bluuebunny/crossref_metadata_2025_split huggingface dataset:
GMSCs suppress osteoclastogenesis and bone erosion in collagen-induced arthritis through curbing NF-KB and RANKL signals (THER5P.900)
Authors: Zheng Song , Gu Jian , Wang Julie , Olsen Nancy 
Published: January 2023

Abstract
Osteoclasts are responsible for bone destruction in rheumatoid arthritis (RA) and gingiva-derived mesenchymal stem cells (GMSCs) can inhibit experimental collagen-induced-arthritis (CIA) model. This study aims to determine if GMSCs also suppress osteolastogenesis and bone erosion in CIA. Osteoclasts were induced from bone-marrow CD11b+ cells with RANKL and macrophage colony-stimulating factor (M-CSF), and assessed with tartrate-resistant acid phosphatase (TRAP) staining. Human osteoclasts were induced from human CD14+ cells. GMSCs were generated and added to cultures with different ratios with CD11b+ or CD14+ cells. NF-κB activation was determined by western blot. 2×106 GMSCs or fibroblast cells were adoptively transferred to DBA1/J mice on day 14 after immunization with CII/CFA. CIA onset and severity were monitored and bone erosion was examined by HE staining. We showed that GMSCs but not fibroblast cells completely suppressed osteoclastogenesis in vitro for human and mice cells. GMSCs injected after immunization and before of onset of CIA significantly suppressed disease development. Treatment with GMSCs dramatically decreased the levels of NF-κB p65/p50 in osteoclasts in vitro and P65/50 and RANKL expression by synovial tissues in vivo. These results suggest that the manipulation of GMSCs may have therapeutic effects on rheumatoid arthritis and other bone erosion related diseases.
Computational modeling of transplant rejection: a systems view of the dynamic roles of T cells, antigen presenting cells, and cytokines during rejection (TRAN1P.941)
Authors: Raimondi Giorgio , Maturo Andrew , Brandacher Gerald , Arciero Julia 
Published: January 2023

Abstract
The quality of life of organ transplant recipients is compromised by complications that arise from ensuing life-long immunosuppression and by limited long-term graft survival due to absence of tolerance induction. There is then an urgent need to better understand the complex interactions of the immune system leading to transplant rejection, so to identify innovative strategies to promote transplant acceptance while preserving immune competence. In this study we have pioneered a systems biology approach in which an experimentally-based mathematical model is used to predict how alterations in the immune response influence the rejection of mouse heart transplants. The model correctly predicts that in the absence of CD4 T cells the transplant survives indefinitely, while in the absence of CD8 T cells it is quickly rejected, providing also dynamics of innate cells that fit recently published data. As expected, decreasing the migration of effectors to the graft delays transplant rejection. This latter relation is not monotonic as increasing the translocation of CD8 T cells has a beneficial effect. This unexpected result derives from the interconnections with the relative concentrations of pro- and anti-inflammatory cytokines and with the ratio between Treg and T cells. This suggests possible key points of intervention for a testable novel strategy of intervention that highlights the usefulness of this innovative multidisciplinary approach.
Aquaporin 4 blockade is a novel strategy targeting ischemia/reperfusion injury in heart transplantation (TRAN1P.942)
Authors: Ayasoufi Katayoun , Kohei Naoki , Farr George , Pelletier Marc , McGuirk Paul , Fairchild Robert , Valujskikh Anna 
Published: January 2023

Abstract
Aquaporin 4 (AQP4) is the major water channel in the brain and its absence reduces brain edema in murine models of stroke.Recently, human and mouse cardiomyocytes were shown to express AQP4. Our goal was to test whether therapeutic inhibition of AQP4 reduces IRI and improves cardiac allografts survival. B6 mice were transplanted with BALB/c heart allografts subjected to 8 h of cold ischemia. Donors received AQP4 inhibitor 0.5h before harvest; hearts were perfused with and stored for 8 h on ice with the inhibitor. Recipients were given AQP4 inhibitor every 6h for 5 days. In the absence of additional immunosuppression, treatment with AQP4 inhibitor significantly prolonged heart allograft survival (MST 2d, n=24 vs. 6 d, n=5), decreased early T cell infiltration and the intragraft expression of IFNγ, granzyme B, perforin, FASL, CXCR3, ICOS, and CXCL10 in comparison to untreated controls. The frequencies of donor-reactive spleen cells secreting IFNγ in recipients treated with AQP4 inhibitor were also four fold lower than those in untreated controls. Blocking AQP4 during allograft storage and early after transplantation prolongs the survival of fully MHC-mismatched heart allografts subjected to 8 h CIS. The graft prolongation is associated with the reduced early memory T cell infiltration into the graft and decreased anti-donor responses. Our data identify AQP4 as a promising therapeutic target to diminish the detrimental effects of prolonged CIS on transplant outcome.
Effector T cell migration to pancreatic islet allografts is dependent on donor antigen recognition and not Gai chemokine signaling. (TRAN1P.945)
Authors: Zhang Qianqian , Dai Hehua , Zeng Qiang , Williams Amanda , Oberbarnscheidt Martin , Lakkis Fadi 
Published: January 2023

Abstract
Host effector T cell graft infiltration is a key step in the pathogenesis of both acute and chronic rejection. Recent data indicate effector T cell migration into primary vascularized organ transplants such as heart and kidney is driven by donor antigen recognition, not via Gai-coupled chemokine receptor signaling. Whether it is true for neovascularized pancreatic islet allografts is unknown. We co-transferred pertussis toxin (Ptx)-treated and untreated TCR-tg (OT-1) effector T cells into B6 recipients 7-10 days after transplanting B6-Act-OVA or B6 islet grafts under the kidney capsule. Ptx irreversibly blocks Gai-dependent signaling. Grafts were imaged 20-24 hours after T cell transfer by two-photon intravital microscopy. We observed B6-Act-OVA grafts were heavily infiltrated with OT-1 effector T cells irrespective of pre-treatment with Ptx or not (453.7+/-170.0 vs. 632.8+/-221.1 per imaging volume, n = 10, p = 0.0522 ), while B6 grafts, which lack the cognate antigen recognized by OT-1 T cells, were free of infiltrate (&lt;5 cells, n = 7). These results indicate that, similar to heart and kidney transplants, islet allograft infiltration with host effector T cells is driven by donor antigen, not by Gai signaling. The paradigm of antigen-driven effector T cell migration is therefore generalizable to both primary vascularized grafts, in which donor antigens are abundantly expressed on the endothelium, and neovascularized cellular grafts whereby the endothelium is of host origin.
Allergic pulmonary inflammation accelerates breast cancer metastasis via increase of myeloid cells in the lung microenvironment (TUM6P.1005)
Authors: Libreros Stephania , Garcia-Areas Ramon , Gazaniga Nathalia , Robinson Philip , Keating Patricia , Iragavarapu-Charyulu Vijaya 
Published: January 2023

Abstract
Inflammation is known to contribute to tumor initiation and metastasis. Since it is not known the effect of pre-existing inflammation on metastasis, we combined a model of allergic pulmonary inflammation with a breast cancer model. Using this model, we found that allergen sensitized mice implanted with mammary tumors had a 5-fold increase in metastatic foci. More importantly allergic tumor bearing mice showed 3X faster tumor cell infiltration in their lungs with accelerated tumor growth and shorter survival. Since macrophages and MDSCs play a key role in the immunosuppressive processes associated with both inflammation and metastasis, we hypothesized that allergic pulmonary inflammatory microenvironment attracts myeloid cells that support incoming tumors cells and accelerates metastasis. We found an increase in the alveolar MO (CD11b-CD11C+CD68+F4/80+Ly6G-), interstitial MO (CD11b+CD11C-CD68+F4/80+Ly6G-), monocytic MDSC (CD11b+Ly6ChighLy6G-F4/80-), and granulocytic MDSC (CD11b+Ly6ClowLy6G+F4/80-) subpopulations in mice with allergic pulmonary inflammation prior to tumor inoculation. These populations were further increased after tumor cell inoculation with the exception the granulocytic MDSCs. Depletion of macrophages and MDSCs prior to tumor cell inoculation in allergen mice decreased tumor growth and metastasis in allergic tumor bearing mice. Targeting specific immune populations could lead to safer and more efficient strategies against breast cancer metastasis.
Impact of MDSC emergence during ovarian cancer in cytokine-deficient mice. (TUM6P.1011)
Authors: Riggs James , DePierri Kelley 
Published: January 2023

Abstract
We have been studying the emergence of myeloid-derived suppressor cells (MDSCs) following i.p. transfer of ID8 cells (mouse ovarian surface epithelial cell carcinoma) into intact, syngeneic, C57BL/6J (wild type, WT) mice. Our current work studies the impact of the cytokine (CK) environment on ID8 expansion and MDSC emergence in the peritoneal cavity (PerC). We wanted to determine if particular pro- (IFNg) or anti- (IL4, IL10) inflammatory CKs were drivers of disease in this model. We found that, relative to WT mice, the development of hemorrhagic ascites was accelerated in IL10-/-, IL4-/-, and IFNgR-/- mice. The greatest percentage and number of MDSCs was observed in IL10-/- mice, which also had the greatest expansion of ID8 cells. ID8 triggered a marked increase in PerC Mfs, particularly in the IL10-/- mice, and lymphocytes, particularly in the IFNgR-/- and IL4-/- mice. However, for all mice, the percent representation of B cells in the total PerC cell pool declined markedly, particularly in the IL10-/- mice (&gt; 70%). Interestingly, beyond the peritoneal locus of disease, both B and T lymphocyte percentages dropped in the spleens of all mice. This was seen without a concomitant increase in the number of MDSCs or appearance of ID8 in the spleen. These observations reinforce that inflammation drives MDSC emergence and suggest that changes in lymphocyte populations could serve as biomarkers for assessing ovarian cancer.
HMGB1 promotes MDSC survival through autophagy (TUM6P.956)
Authors: Parker Katherine , Ostrand-Rosenberg Suzanne 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSC) are immune suppressive cells that are elevated in most individuals with cancer where their accumulation and suppressive activity are driven by inflammation. Because MDSC globally inhibit anti-tumor immunity, we are determining how their turn-over is regulated. We have established that the damage-associated molecular pattern molecule (DAMP) High Mobility Group Box protein 1(HMGB1) is critical for MDSC accumulation and function, and is ubiquitously present in the tumor microenvironment. Studies by others demonstrated that HMGB1 is induced by reactive oxygen species (ROS) and facilitates tumor cell survival by inducing autophagy. These findings have led us to hypothesize that HMGB1 regulates MDSC turn-over by propagating a pro-autophagic state that is self-sustained by the high levels of ROS produced by MDSC. Since autophagy involves translocation of HMGB1 from the nucleus to the cytosol, we have tested this hypothesis by starving MDSC and inhibiting HMGB1 translocation into the cytosol, and by inhibiting MDSC production of ROS. Inhibition of HMGB1 translocation limited MDSC autophagy and decreased MDSC survival, while inhibition of ROS reduced autophagy in MDSC. These results suggest that HMGB1 prolongs MDSC survival by inducing a pro-autophagic state in MDSC.
Inhibiting fatty acid oxidation blocks the immunosuppressive functions of myeloid-derived suppressor cells and enhances cancer therapies (TUM6P.960)
Authors: Hossain Fokhrul , Al-Khami Amir , Wyczechowska Dorota , Hernandez Claudia , Zheng Liqin , Reiss Krzystoff , Del Valle Luis , Trillo-Tinoco Jimena , Rodriguez Paulo , Ochoa Augusto 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSC) promote tumor growth by inhibiting T cell immunity through various mechanisms including the depletion of arginine and the production of nitric oxide and peroxinitrites. Blocking of any single mechanism has only moderate therapeutic effects because MDSC show high adaptability and upregulate the remaining suppressive pathways. We studied whether disrupting the energy metabolic pathway(s) of MDSC would modulate their immunosuppressive functions and enhance antitumor effects. Using several murine tumor models, we found that tumor-infiltrating MDSC (T-MDSC) increased fatty acid uptake and activated fatty acid oxidation (FAO). This was accompanied by increased mitochondrial mass and function, key FAO enzymes, and oxygen consumption rate. These metabolic changes paralleled with the induction of immunosuppressive mechanisms. Inhibition of FAO blocked immunosuppressive mechanisms and functions in T-MDSC and decreased their production of immunosuppressive cytokines. Interestingly, it also diminished the number of regulatory T cells and resulted in a T cell-dependent decrease in tumor growth. FAO inhibition also significantly increased antitumor effects with low-dose chemotherapy or cellular immunotherapy. Human MDSC demonstrated similar characteristics. These data support the possibility of testing FAO inhibition as a novel approach to block MDSC and enhance various cancer therapies. *F. Hossain and A.A. Al-Khami contributed equally to this work.
Genetic depletion of cyclin dependent Kinase-5 results in enhanced tumor rejection via a CD4+ T cell dependent mechanism (TUM7P.1023)
Authors: Dorand Rodney , Petrositue Agne , Nthale Joseph , Scrimieri Francesca , Pareek Tej , Myers Jay , Barkauskas Deborah , Letterio John , Stearns Duncan , Huang Alex 
Published: January 2023

Abstract
CDK5 is a non-canonical member of the CDK family that does not require cell cycling for activity but relies on binding to its obligate activators, p35 and p39 which are mostly expressed in post mitotic neurons. CDK5 is required for normal Central Nervous System (CNS) development. Medulloblastoma (MB), derived from primitive cerebellar granule neuron precursors, is the most common pediatric CNS malignancy. Utilizing a short hairpin RNA approach to silence Cdk5 expression in a C57BL/6 (B6) syngenic MB line derived from p53+/- PTCH-/- (MM1), we found that 19 days after subcutaneous inoculation, mice with silenced Cdk5 (MM1-CDK5-KD) revealed a 78% tumor free survival (N=36) compared to 4% (N=24) of mice inoculated with wild type MB (MM1-WT). Furthermore, when injected into Nu/Nu mice, MM1-WT and MM1-CDK5-KD had similar subcutaneous growth kinetics. When CD4+ cells were depleted in B6 mice inoculated with MM1-CDK5-KD tumor free survival decreased to 28% (N=15). We sought to interrogate the role of the immune system in enhanced killing of MM1-CDK5-KD tumors. When exposed to IFN-γ stimulation in vitro, genetic profiling of MM1-CDK5-KD revealed a decrease in PD-L1 expression, a known immune checkpoint surface molecule, compared to MM1-WT. These results indicate a functional role for Cdk5 in MB immune evasion which, when circumvented via Cdk5 inhibition, leads to increased susceptibility to anti-tumor surveillance.
Diversity index of mucosal resident T lymphocyte repertoire predicts clinical prognosis in gastric cancer (TUM7P.1028)
Authors: Wan Ying , Jia Qingzhu , Zhou Junfeng , Chen Gang , Shi Yan , Yu Haili , Lin Regina , Jiang Ning , Yu Peiwu , Li Qi-Jing 
Published: January 2023

Abstract
A characteristic immunopathology of human cancers is the induction of tumor antigen-specific T lymphocyte responses within solid tumor tissues. Current strategies for immune monitoring focus on the quantification of the density and differentiation status of tumor-infiltrating T lymphocytes; however, properties of the TCR repertoire ‒ including antigen specificity, clonality, as well as its prognostic significance ‒ remain elusive. In this study, we enrolled 28 gastric cancer patients and collected tumor tissues, adjacent normal mucosal tissues, and peripheral blood samples to study the landscape and compartmentalization of these patients’ TCR beta repertoire by deep sequencing analyses. Our results illustrated antigen-driven expansion within the tumor compartment and the contracted size of shared clonotypes in mucosa and peripheral blood. Most importantly, the diversity of mucosal T lymphocytes could independently predict prognosis, which strongly underscores critical roles of resident mucosal T-cells in executing post-surgery immunosurveillance against tumor relapse.
Imaging in cancer immunology: phenotyping of multiple immune cell subsets in-situ in FFPE tissue sections (TUM7P.1029)
Authors: Mansfield James , Hoyt Clifford , Stack Ed , Feldman Michael , Bifulco Carlo , Fox Bernard 
Published: January 2023

Abstract
There has been a rapid grown in the field of tumor immunobiology in recent years as a result of recent successes in cancer immunotherapies, and it is clear that immune cells play many sometimes conflicting roles in the tumor microenvironment. However, obtaining phenotypic information about the various immune cells in and around the tumor has been a challenge. Existing methods can deliver phenotypic information on homogenous samples (e.g., flow cytometry or PCR) or morphologic information in single stain IHC. We present here a methodology for delivering quantitative per-cell marker expression and phenotyping, analogous to that obtained from flow cytometry, but from cells imaged in situ in FFPE tissue sections. This methodology combines: the sequential multi-marker labeling of up to 6 antigens using antibodies all of the same species; automated multispectral imaging to remove problematic FFPE tissue autofluorescence and correct cross-talk between fluorescent channels; and an automated analysis that can quantitate the per-cell marker expression, determine the cellular phenotype, count these cells separately in the tumor compartment and in the stroma and provide high-resolution images of their distributions. We will show a 6-plex assay in breast cancer showing the application of the multiplexed staining, per-cell quantitation and cellular phenotyping in FFPE tissue sections, as well as methods to explore the spatial distributions of the phenotyped cells in and around the tumor.
STING protects against colitis-associated colorectal tumorigenesis by modulating intestinal inflammation (TUM7P.1018)
Authors: Zhu Qifan , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
Stimulator of IFN genes (STING) is an innate immune sensor for cytosolic cyclic dinucleotides and also serves as an adaptor molecule for a number of intracellular DNA sensors. While STING has been shown to be critically involved in modulating immune responses during infections and autoimmune diseases, its physiological role in cancer remains largely unknown. In this study, we demonstrated that STING-deficient mice are highly susceptible to colitis-associated colorectal cancer with overt intestinal proliferation during early stages of tumorigenesis. Hyperactive NF-kB and STAT3 signaling was responsible for increased levels of proinflammatory cytokines observed in STING-deficient mice. Furthermore, intestinal tissues deficient in STING exhibited elevated level of DNA damage maker, γH2AX and a defect in caspase-1-mediated colonic IL-18 release, highlighting its fundamental functions in maintaining genomic integrity and activating inflammasome. Therefore, our results identify an unexpected and important role for STING in mediating protection against colorectal tumorigenesis.
Epigenetic silencing of Th1-type chemokines dictates effector T cell tumor trafficking and predicts patient outcome (TUM7P.1019)
Authors: Peng Dongjun , Kryczek Ilona , Nagarsheth Nisha , Zhao Lili , Wei Shuang , Zhao Ende , Vatan Linda , Szeliga Wojciech , Kotarski Jan , Tarkowski Rafal , Dou Yali , Cho Kathleen , Liu Rebecca , Zou Weiping 
Published: January 2023

Abstract
Epigenetic silencing including histone modifications and DNA methylation is an important tumorigenic mechanism in cancer. However, its role in cancer immunopathology and immunotherapy is poorly understood. In this study, we focused on the infiltration of effector T lymphocytes into ovarian tumor and established the histone H3 lysine 27 trimethylation (H3K27me3) and DNA hypermethylation as epigenetic silencing mechanisms regulating Th1-type chemokines CXCL9 and CXCL10 production. We found that a polycomb group (PcG) protein, enhancer of zeste 2 homolog (EZH2)-based histone H3 lysine 27 trimethylation (H3K27me3) alone or in collaboration with DNA methyltransferases (DNMTs) selectively control the tumor production of CXCL9 and CXCL10, and subsequently determine effector T cell trafficking to the tumor microenvironment. Treatment with epigenetic modulators removes the repression and increases effector T cell tumor infiltration, slows down tumor progression, and improves therapeutic efficacy of PDL1(B7-H1) checkpoint blockade and adoptive T cell transfusion. Moreover, tumor EZH2 and DNMT1 are negatively associated with tumor infiltrating CD8+ T cells and patient outcome. Our results indicate that epigenetic silencing of Th1-type chemokine is a novel tumor immune evasion mechanism. Selective epigenetic reprogramming alters T cell landscape in cancer and enhances clinical efficacy of cancer therapy.
IL-2 complex rescues CD8+ T cell tolerance for cancer immunotherapy (VAC7P.1041)
Authors: Swier Lauryn , Berrien Melissa , Teague Ryan 
Published: January 2023

Abstract
IL-2 is a well-characterized T cell growth factor that promotes proliferation and effector function of CD8+ T cells. For this reason, IL-2 has been used to boost immune responses in patients with melanoma. However, IL-2 treatment requires large potentially toxic doses of IL-2, and often only limited therapeutic efficacy is acheived. Efforts to enhance the efficacy and safety of IL-2 have recently focused on the use of cytokine/antibody complexes. These complexes demonstrate greater bioactivity in vivo relative to IL-2 alone, making them an attractive approach for improved immunotherapy. One of the major impediments to anti-tumor immunity is CD8+ T cell tolerance, where tumor-reactive T cells are either eliminated by apoptosis or rendered dysfunctional/anergic. The influence of IL-2 complex (IL-2c) on tolerant CD8+ T cells has not been explored. We utilized a mouse model of T cell tolerance to examine how IL-2c treatment impacts tolerant tumor-reactive T cells during immunotherapy for leukemia. Specifically, IL-2c was administered daily starting 3 days after T cell transfer into tumor-bearing mice. The function and persistence of transferred tumor-reactive T cells was assessed 5 days later. Treatment with IL-2c prevented apoptotic deletion of transferred CD8+ T cells and improved effector function. These results suggest that IL-2c treatment is sufficient to overcome peripheral T cell tolerance, and support further exploration of IL-2c as an immunotherapeutic for cancer.