bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhancement of neutrophil signaling and effector function by Imprime PGG in response to tumor cells in biomarker positive individuals (TUM7P.937)
Authors: Leonardo Steven , Gorden Keith 
Published: January 2023

Abstract
Imprime PGG is a yeast β-glucan immunomodulator being developed for tumor therapy. Imprime PGG, in combination with tumor-specific monoclonal antibodies (MAb), is more effective in human trials than MAb therapy alone. Imprime PGG’s mechanism of action involves redirecting the innate immune system, in particular neutrophils, to kill MAb-targeted tumors cells. Recent work has identified a subset of individuals with high anti-β-glucan antibodies (ABA) that demonstrate greater efficacy to Imprime PGG treatment in vivo. Using ABA levels, we investigated signaling and functional differences in Imprime PGG treated neutrophils between biomarker positive and negative individuals. In this study we developed a calcium flux assay to assess Imprime PGG’s ability to modulate neutrophil signaling in response to MAb-targeted tumor cells. Our data demonstrates that ABA high individuals show a 3-fold greater enhancement in calcium signaling then ABA low individuals after Imprime PGG treatment. Functional activity was measured by Fc-mediated oxidative burst in response to antibody coated particles; a mechanism mimicking how antibody-coated tumor cells are recognized and eradicated by Imprime PGG treated neutrophils. ABA high individuals demonstrated a 5-fold greater enhancement in Fc-mediated oxidative burst upon Imprime PGG treatment then ABA low individuals. Our results demonstrate that Imprime PGG modulates signaling and functional activities in neutrophils of biomarker positive individuals.
MUC1 modulates NF-kB-dependent gene expression of pro-inflammatory cytokines by altering their epigenetic regulation (TUM7P.944)
Authors: Cascio Sandra , Sciurba Joshua , Finn Olivera 
Published: January 2023

Abstract
MUC1 is a transmembrane glycoprotein overexpressed and hypoglycosylated in human adenocarcinomas as well as sites of chronic epithelial inflammations. The MUC1 extracellular domain plays an important role in cell-cell communication, protein-protein interaction and signaling pathway activation. We investigated the role of MUC1 in transcriptional regulation of pro-inflammatory cytokines, and whether NF-kB signaling pathway is required. Our results showed that under TNFα treatment the entire molecule MUC1 is translocated to the nucleus and binds cytokine promoters. In particular, ChIP and Real-time PCR experiments revealed that MUC1, in association with p65, up-regulated IL6 and TNFα expression by binding to their promoter regions in both mouse and human cancer cells. Because proteomic analysis showed that MUC1 is able to interact directly with histone proteins, we hypothesized that MUC1 might influence the transcriptional activity of cytokines by inducing epigenetic modification. Consistent with other studies revealing the cross-talk between histone acethylation, methylation and ubiquitination, our preliminary data demonstrated that full length form of MUC1 modulates the expression of several factors involved in chromatin remodeling such as CBP/p300, HDAC1, ubH2A and EZH2. We also found that MUC1 controls their interaction with IL6 and TNFα promoters in breast cancer cells. This is an important finding that may explain the mechanism by which MUC1 regulates cancer progression.
Immunization of CD200R1KO mice post surgical resection of EMT6 breast cancer leads to long-term (&gt;1year) cure of metastatic tumor growth (TUM7P.921)
Authors: Gorczynski Reg , Chen Zhiqi , Khatri Ismat , Podnos Anna 
Published: January 2023

Abstract
Attenuated expression of CD200 decreases local tumor growth and metastasis in immunocompetent animals. We explored the ability to cure BALB/c CD200KO or CD200R1KO mice of tumors ≤1cm3 in size by surgical resection of localized tumor, followed by immunization with irradiated EMT6 cells along with CpG as adjuvant. Control animals developed pulmonary and liver metastases within 30 days of surgery. Protection was seen in both CD200KO or CD200R1KO mice, with no macroscopic lung/liver metastases observed in CD200R1KO mice on sacrifice at day 300. After resection and immunization, draining lymph nodes from controls contained tumor cells cloned at limiting dilution in vitro even before pulmonary and hepatic metastasis was seen. Within the limits of detection of the assay used (sensitivity ~1 in 107 cells) no tumor cells were detected at limiting dilution in similarly treated CD200R1KO mice, and significant reductions were seen in CD200KO mice. Infusion of anti-CD4, but less so anti-CD8, mAb into surgically treated and immunized CD200R1KO mice attenuated protection from both macroscopic (liver/lung) and microscopic (assayed by limiting dilution of DLN) metastasis. Adoptive transfer of lymphocytes from treated CD200R1KO mice to surgically-treated control mice also attenuated metastatic growth of tumor, which was abolished by pretreatment of transferred cells with anti-CD4 mAb. Our data suggest CD200:CD200R attenuates a potentially tumor-protective CD4 host response to breast cancer.
Necrosis associated factors (S100A4 and uric acid) promote accumulation of MSCs which induce IL10+/IDO+ lymphocytes (TUM7P.949)
Authors: Lotfi Ramin , Eisenbacher Judith , Jahrsdörfer Bernd , Kaltenmeier Christof , Rojewski Markus , Yildiz Tatjana , Beyer Thamara , Erle Alexander , Wiegmann Denis , Grassl Stefan , Körper Sixten , Wiesneth Markus , Schrezenmeier Hubert 
Published: January 2023

Abstract
Necrosis with subsequent release of damage-associated molecular patterns (DAMPs) is a characteristic feature of advanced solid tumors. DAMPs impact the tumor-microenvironment stimulating tumor-associated cells like mesenchymal stromal cells (MSCs). Tumor-infiltrating-MSCs are associated with tumor progression and metastasis. Oxidized DAMPs lose their stimulatory capacity on MSCs. As a DAMP-member S100A4 is sensitive to oxidation while uric acid (UA) acts as an antioxidant. We tested these two DAMP-members separately and in combination for their biologic activity on MSCs. S100A4 and UA showed a dose-dependent chemotactic activity on MSCs with synergistic effects when both DAMPs were combined. Substituting for UA, alternative antioxidants (Vitamin-C, dithiothreitol, and acetylcystein) had also a comparable synergistic effect on the chemotactic activity of S100A4, emphasizing the reducing potential of UA being responsible for the observed synergy. Yet, S100A4 and UA inhibited MSC proliferation without impacting their viability. Nevertheless, in the presence of S100A4 or UA, MSCs gained immunosuppressive capacities by inducing IL-10 and indoleamine dioxygenase expressing lymphocytes. We characterized S100A4 and UA as necrosis-associated factors playing a crucial role within MSC biology and thus immunoregulation. Our results have implications for therapeutic approaches targeting the tumor microenvironment and addressing the immunosuppressive nature of necrotic tumor.
The role of intestinal epithelial exosomes in the progression of colon cancer (TUM7P.952)
Authors: Deng Zhong-bin , Wang Baomei , Zhuang Xiaoying , Jiang Hong , Wang Qinglong , Zhang Lifeng , Zhang Huang-Ge 
Published: January 2023

Abstract
Accumulating evidence indicates that dysregualtion of host-microbiota interaction in gut is associated with the development of colon cancer and with chronic inflammation. This chronic inflammation results in part from the interaction between inflammatory cells and gut microbiota, but pathway(s) leading to gut microbial mediated inflammation, and promoting colon cancer were heretofore unknown. Here we demonstrate that exosomes that bud from colon tumor cells play a key role in the recruitment of inflammatory CCR6+CD4+Th17+ into tumor site and subsequently promote tumor growth. We found that administered tumor exosomes can induce spontaneous intestinal polyposis in the Apc (Min/+) mouse model, a widely used spontaneous colorectal cancer model for human familial adenomatous polyposis (FAP) in a CCL20 dependent manner. We further demonstrate that bacterial products up taken by tumor cells promote the production of PGE2+CCL20+ tumor exosomes. Our studies identify a key mechanism of tumor exosomes-mediated T cells homing to intestine.
IRF5 is a novel regulator of CXCL13 expression in breast cancer that increases CXCR5+ B and T cell trafficking to the tumor (TUM7P.955)
Authors: Pimenta Erica , De Saurav , Feng Di , Hall Kelly , Ran Sophia , Barnes Betsy 
Published: January 2023

Abstract
Clinical studies using predictive signatures have shown that a high immune signal from whole tumor expression profiles correlate with improved outcome in some cancers, including breast cancer. Factors regulating immune cell trafficking to the tumor, however, are not known. Utilizing MDA-MB-231 breast cancer cells grown in 3D culture, we examined whether intratumoral expression of interferon regulatory factor 5 (IRF5) regulates the profile of cytokines/chemokines expressed. Previous work has shown that IRF5 expression is lost in ~80% of all invasive ductal carcinomas examined. Using focused arrays, we found a number of cytokines/chemokines that were dysregulated between IRF5-positive and -negative MDA-MB-231 cells. CXCL13 was identified as a direct target of IRF5. Using tumor-conditioned media (TCM) from IRF5-positive and -negative cells, we found a significant increase in B and T cell trafficking to TCM from IRF5-positive cells. Subset analysis revealed that primary CXCR5+ B and T cells showed significantly reduced migration to IRF5-negative TCM or IRF5-positive TCM depleted of CXCL13. CXCR5 is the primary receptor for CXCL13. Analysis of human breast tumor tissues revealed a positive correlation between IRF5 and CXCL13 expression providing clinical relevance to this study. Data support that IRF5 directly regulates a network of genes that shapes a tumor immune response and may serve as a novel prognostic marker for chemotherapy and immunotherapy response.
NLRX1 attenuates tumorigenesis through the negative regulation of AKT and NF-κB signaling (TUM7P.924)
Authors: Allen Irving , Simmons Alysha , Capria Vittoria , LeRoith Tanya , Robbins Greg , Heid Bettina , Philipson Casandra , Dervisis Nikolaos , Yuzbasiyan-Gurkan Vilma , Hontecillas-Magarzo Raquel , Bassaganya-Riera Josep , Ting Jenny 
Published: January 2023

Abstract
Members of the NLR family of pattern recognition receptors are essential mediators of the host immune response. Recently, our lab and others identified a novel sub-group of NLRs that function as negative regulators of inflammation. One of the members of this sub-group, NLRX1, is a potent regulator of interferon, NF-κB and autophagy signaling. Each of these signaling pathways has been linked to diverse types of cancer. Thus, we hypothesized that NLRX1 attenuates tumorigenesis through the negative regulation of overzealous innate immune system signaling. To evaluate this hypothesis, we utilized a urethane based model of tumorigenesis and explored disease progression in Nlrx1-/- mice. Mice lacking NLRX1 exhibited dramatically increased morbidity and mortality and were found to develop a rare neoplasia, diagnosed as histiocytic sarcoma. Of the already rare histiocytic disorders in humans, histiocytic sarcoma is exceedingly uncommon and the rarity of this disease has resulted in a paucity of data pertaining to diagnostic and therapeutic options. Mechanistically, our data indicates that NLRX1 attenuates tumorigenesis through the negative regulation of AKT and NF-κB signaling. These pathways were found to be associated with macrophage hyperproliferation in the absence of NLRX1. Our data extends the function of NLRX1 beyond its currently characterized role in host-pathogen defense and reveals a significant contribution for this NLR in cancer pathobiology.
A novel protein from lamprey immune body with selective cytocidal actions to tumor cells (TUM7P.925)
Authors: Li Qingwei , Pang Yue , Wang Shiyue , Ba Wei , Yu Tao , Liang Hongfang , Bi Dan 
Published: January 2023

Abstract
We purified and identified a novel protein, derived from cells of immune body tissues in the adult lamprey, the protein has strong cytocidal activity against various human cells with a markedly divergent target specificity, recognizes and kills human breast adenocarcinoma MCF-7, human caucasian chronic myelogenous leukaemia k562 cancer cells as well as some classes of human tumor cells, and less cytotoxic to the normal tissue cells. The novel protein induced remarkable morphological alterations, cell blebbing, cytoskeletal alterations, mitochondrial and endoplasmic reticulum fragmentation. Then the plasma membrane permeability was increased immediately and most of the cytoplasmic proteins and organelle proteins released into medium from tumor cells after the protein treatment. The novel protein shows specific binding to cell membrane and requires the high content of phosphatidylserine for the effective cytolysis. To confirm the novel protein effect on tumor, tumor xenograft experiments were performed, these results confirm that the novel protein inhibit tumor growth in vivo. Taken together, these observations suggest the novel protein play a crucial role in tumor cells survival and growth.
Lymphatic-targeted liposome vaccines promote long-term immunological memory (VAC10P.963)
Authors: Ma Yifan , Wang Ce , Liu Peng , Cai Lintao 
Published: January 2023

Abstract
Lymphatic-targeted vaccine delivery with liposome formulations has been reported to enhance the immunogenicity of vaccines. However, its effect on long-term immunological memory remains unclear. In the present study, OVA vaccines were formulated by cationic liposomes with different surface modification: DOTAP liposomes (LP), PEGylated DOTAP (LP-PEG) liposomes, and mannosylated DOTAP (LP-Man) liposomes. The in vivo imaging showed that LP mostly accumulated near the injection site, indicating its depot effect. In contrast, LP-PEG and LP-Man liposomes effectively accumulated in draining lymph nodes and the spleen, indicating their lymphatic targeting. More importantly, different liposomes differentially regulate antibody response and B-cell memory. Although LP vaccine led to the highest level of anti-OVA IgG among three liposome formulations on day 40 post priming, they failed to establish an effective memory response. In contrast, lymphatic-targeted LP-PEG and LP-Man vaccines elicited sustained antibody response and robust recall responses 3 months after priming. The enhanced long-term immunological memory by lymphatic-targeted liposome vaccines could be attributed to vigorous and persistent germinal center response, as well as increased Tfh cells and central memory CD4+ T cells in the spleen and lymph nodes. Hence, lymphatic-targeted vaccine delivery with surface modified cationic liposomes could be an effective strategy to promote long-lasting immunological memory.
Th1-promoting adjuvanticity of cholera toxin depends on Gsα in CD11b+ dendritic cells (VAC10P.969)
Authors: Lycke Nils , Mattsson Johan , Yrlid Ulf 
Published: January 2023

Abstract
Despite an extensive literature on the mechanisms of action of cholera toxin (CT) we still lack critical information about how CT augments immune responses. Although several studies have identified dendritic cells (DC) as critical target cells for the adjuvant function there is still not a distinct subset of DCs that has been shown to convey the adjuvant effect. Even more confusing is the fact CT has been considered a strong Th2-skewing adjuvant, but many researchers have found potent IFNγ and Th1 promoting function, despite that CT interferes with IRF8 and blocks IL-12 production. We immunized wild-type, Batf3-/- (lacking CD8α+ DCs) and Cre-lox mice with a CD11c -selective defect in Gsα, the membrane target protein for CT’s ADP-ribosylation. While CT exerted extensive global effects on secondary lymphoid tissues with recruitment and redistribution of both innate and adaptive immunocytes leading to a complete loss of CD4 T cell priming ability, this condition was effectively restored by transferring freshly isolated CD11c+ DCs. As Batf3-/- mice exhibited intact adjuvanticity, CD8α+ DCs were not required for the adjuvant effect. However, we found that CD11b+CD11c+ cells were absolutely needed and that Gsα-expression was critical. CD4 T cell priming was unrestricted and Th1, Th2 and Th17 differentiation was observed also in the absence of IL-12, arguing that CT exerts an adjuvant effect independent of IRF8.
Chimeric-antigen receptor engineered human T lymphocytes from induced pluripotent stem cells for cancer therapy. (VAC11P.1003)
Authors: Themeli Maria , Sadelain Michel 
Published: January 2023

Abstract
We hypothesized that genetic engineering of induced pluripotent stem cells (iPSC) with Chimeric Antigen Receptors (CARs) would be an efficient strategy to generate readily available functionally enhanced T cells targeted to a tumor antigen of interest. Since combined CD28 and 4-1BB signaling promote T cell proliferation and survival, we genetically engineered iPSCs from peripheral blood lymphocytes (T-iPSC) to express 1928z, a second-generation CAR specific to the B-cell lineage antigen CD19, and 4-1BBL. Stably transduced 1928z/4-1BBL-T-iPS lines were then directed to differentiate in vitro into CD3+TCRαβ+ T cells that expressed the 1928z CAR and 4-1BBL on the surface (1928zBBL-TiPSC-T). Upon exposure to cells expressing CD19, 1928zBBL-TiPS-T cells rapidly responded by forming clusters, increased the expression of activation markers and secreted type 1 cytokines. We were able to expand 1928zBBL-TiPSC-T cells and expanded 1928zBBL-T-iPSC-T cells displayed high antigen-specific cytotoxic activity in vitro. Treatment of CD19+ tumor-bearing mice with 1928zBBL-TiPSC-T cells delayed tumor progression and resulted in a significant survival advantage. In summary, the combination of iPSC and CAR technologies we describe here offers a tantalizing new source of “off the shelf” T cells of predetermined antigen specificity and enhanced properties.
Hyper-IL15 enhances human immune function and suppresses hepatitis B virus replication in humanized mice with both human liver and immune system (VAC12P.1024)
Authors: Cheng Liang , Li Feng , Bility Moses , Chi Liqun , Su Lishan 
Published: January 2023

Abstract
HBV infection remains a leading cause of death worldwide. However, the lack of robust small animal model has hindered the progress in hepatitis B virus research and the development of new treatment options. Immunodeficient mice reconstituted with both human immune system and liver provide us a powerful in vivo model to study HBV pathogenesis and evaluate potential therapeutics. Cytokines targeting T cells and NK cells are attractive clinical interventions against viruses. Here we tested the therapeutic effect of human hyper-IL15(IL15 fused with IL15Rα sushi domain) on HBV replication in vivo in our humanized mice with both human liver and immune system. We showed that hydrodynamic injection of HBV1.3 expression plasmid led to persistent HBV replication for at least 4 months in humanized mice. Ectopic expression of hyper-IL15 in humanized mice one week after HBV plasmid injection significantly inhibited HBV replication. The HBV inhibition effect was correlated with increasing human T cell and NK cell numbers and activation in humanized mice. To further test the therapeutic potential of hyper-IL15 on chronic phase of hepatitis B, we treated the humanized mice with hyper-IL15 7.5 weeks after HBV persistence in vivo. Our data showed that the HBV genome was undetectable in 75% of hu-mice 3 weeks after hyper-IL15 treatment. Our encouraging results warrant future exploration of the use of hyper-IL15 as an immunotherapy reagent to treat chronic hepatitis B in humans.
Therapeutic vaccination against tumor pericyte-associated antigens DLK1 and DLK2 (VAC12P.1013)
Authors: Fabian Kellsye , Chi-Sabins Nina , Storkus Walter 
Published: January 2023

Abstract
The tumor microenvironment (TME) promotes unique epigenetic programming of stromal cell populations, including vascular endothelial cells (VEC) and pericytes, leading to differential expression of antigens that may be recognized by therapeutic T cells. These antigenic differences may be targeted immunologically via specific vaccination. We show that pericytes in human and murine renal cell carcinoma (RCC) and melanoma differentially overexpress the antigen Delta-like homolog 1 (DLK-1). DLK-1 is an EGF-like protein that functions as a Notch pathway antagonist. We demonstrate that both peptide and genetic vaccination of RENCA-bearing BALB/c mice against DLK-1 results in tumor growth inhibition and removal of DLK-1 within the TME. However, DLK-1 removal also led to a compensatory increase in DLK-2 expression (a DLK-1 homolog and NOTCH antagonist) in the murine RENCA TME. Using recombinant lentiviruses encoding murine DLK-1 and DLK-2, we show that coordinate genetic vaccination against these two antigens leads to superior therapeutic inhibition of RENCA tumor growth compared to vaccination against either DLK-1 or DLK-2 alone. The coordinate vaccination also resulted in the activation of DLK-1- and DLK-2-specific CD8+ T cells, recruitment of T cells into the TME and normalization of tumor blood vessels. We conclude that vaccination against DLK-1 and/or DLK-2 may represent an effective strategy for vascularized, solid forms of cancer, including RCC and melanoma, among others.
NK cell activation by the heat shock protein gp96 (VAC12P.1018)
Authors: Sedlacek Abigail , Binder Robert 
Published: January 2023

Abstract
Heat Shock Proteins (HSPs) are key mediators in the development of immune responses. Following immunization with HSPs chaperoning tumor antigens, anti-tumor immune responses are elicited. The current model of HSP mediated anti-tumor immunity suggests HSPs bind to and are internalized by antigen presenting cells (APCs), which results in the priming of effector T cells that subsequently inhibit tumor growth. In addition to APCs and T cells, NK cells have also been demonstrated to be essential for HSP mediated anti-tumor immunity. However what role NK cells play in eliciting immunity is poorly understood. In these studies, we aim to characterize the activation of NK cells following stimulation with the prototypical HSP, gp96. Our results show that, following incubation with gp96 in vitro, we observe an increase in the frequency of NK cells containing the intracellular effector proteins granzyme B and IFNγ. Although intracellular levels of IFNγ are increased, we do not detect IFNγ secretion and the intracellular levels are less than what is observed following cytokine mediated activation. Due to these responses, we are currently investigating NK cells for the presence of the HSP receptor CD91. Collectively, these data indicate that directly, HSP stimulation results in partial NK cell activation, and other mechanisms, such as indirect activation via HSP activated APCs, may be necessary to observe optimal NK cell effector functions.