bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel p65 inhibitor suppresses glutamate release and proliferation of macrophages: therapeutic implications for chronic neurological diseases. (THER3P.887)
Authors: Srinivasan Mythily , Blackburn Corinne , Lahiri Debomoy 
Published: January 2023

Abstract
Activated nuclear factor-κB (NFκB) plays critical role in the pathogenesis of chronic neurological diseases constituting an attractive therapeutic target. Based on the cross talk between NFκB and glucocorticoid signaling pathways, we developed a novel strategy to selectively target p65, the functionally critical subunit of NFκB. Glucocorticoid induced leucine zipper (GILZ) is a “divergence” hub linked to multiple proteins in the NFκB signaling pathway. Structurally, proline rich carboxy terminus of GILZ binds the transactivation domain of p65. An immunomodulatory peptide derived from the p65 binding region of GILZ, the GILZ-P ameliorated autoimmune disease in mice. Here we determine the secondary structural characteristics and investigate the efficacy of GILZ-P in lipopolysaccharide (LPS) activated ThP-1 macrophages used as model systems for human microglial cells. GILZ-P treatment did not adversely affect cell viability as determined by vital staining and flow cytometry. LPS stimulated mature macrophages treated with GILZ-P exhibited reduced proliferation and inflammatory cytokine secretion as assessed by the MTT assay and ELISA respectively. Mechanistically the inhibitory potential of GILZ-P was substantiated by the reduced nuclear translocation of p65 and decreased glutamate release. Collectively, our data support anti-inflammatory and neuroprotective effects for the PPII helical GILZ-P suggesting therapeutic applications in neuroinflammatory-neurodegenerative conditions.
Cbl-b deficiency results in abnormalities in both T cell sphingosine-1-phosphate receptor 1 function and response to the multiple sclerosis therapeutic agent, FTY720 (THER3P.881)
Authors: Fujiwara Mai , Anstadt Emily , Khanna Kamal , Clark Robert 
Published: January 2023

Abstract
FTY720 is a sphingosine-1-phosphate receptor 1 (S1P1) modulator used to treat Multiple Sclerosis (MS). While FTY720 is postulated to mediate its effect by enhancing T cell S1P1 internalization and degradation, the mechanisms underlying the therapeutic effect of FTY720 in MS are not fully understood. To further clarify the functional effects of FTY720 we utilized Cbl-b-deficient (Cbl-b-/-) mice. Cbl-b is an E3 ubiquitin ligase that regulates the PI3K-Akt pathway in T cells and its absence leads to resistance to regulatory T cells (Tregs) and multi-organ autoimmunity in mice. Given that MS has been associated with both SNPs in the CBLB gene and resistance to Tregs, Cbl-b-/- mice represent an important model for studying MS. We now report that Cbl-b-/- T cells demonstrate multiple functional abnormalities in S1P1. These include significantly less lymphopenia induced by the S1P lyase inhibitor, THI, (Cbl-b-/- blood CD4+ T cells decreased 19% vs 59% in wild type (WT) at 48 hrs), suggesting abnormal regulation of Cbl-b-/- T cell S1P1 in response to the endogenous ligand S1P. Paradoxically, Cbl-b-/- T cells demonstrate enhanced lymphopenia induced by the therapeutic agent FTY720 (Cbl-b-/- blood CD4+ T cells decreased 51% vs 26% in WT at 120 hrs). These results for the first time identify a complex role for Cbl-b in regulating expression and turnover of T cell S1P1 and suggest that SNPs in the CBLB gene may associate with a differential response to FTY720 in MS patients.
Senescence associated inflammasome (SAI) activation promotes cancer progression (TUM4P.909)
Authors: Guo Beichu , Zhang Jinyu 
Published: January 2023

Abstract
Oncogene-induced senescence functions as a barrier against cell transformation and tumorigenesis. Paradoxically, recent studies indicate that senescence also promotes tumor progression through secretion of inflammatory molecules or senescence-associated secretory phenotype (SASP). However, the signaling pathways leading to SASP and the interaction between senescent cells and innate immune cells are poorly understood. Utilizing human and murine breast cancer cells, as well as breast cancer mouse models, this study demonstrates that senescence-associated inflammasome (SAI) generate a positive feedback loop to propagate inflammation through interaction with innate immune cells, contributing to the tumor development. Our data show that cellular senescence induced by oncogenes, such as K-Ras12V and Her2/ERBB2, led to inflammasome activation and IL-1beta production in both murine and human breast cancer cells. Notably, we found that blocking inflammasome activity could suppress both IL-1beta production and oncogene-induced senescence in breast cancer cells. Our results further show that the crosstalk between senescent cells and innate immune cells promoted inflammation and breast cancer progression. Together, our findings demonstrate a critical role for the senescence-associated inflammasome in oncogenic inflammation and tumor development.
The myeloid receptor TREM2 exacerbates DSS-induced colitis and promotes colitis-associated cancer (TUM4P.916)
Authors: McVicar Daniel , Howard O.M.Zack , Sublesk Jeff , Quigley Laura , Yang Youfeng , Rayman Patricia , Rini Brian , Linehan W. Marston , Sayers Thomas , Finke James , Stappenbeck Thaddeus , Ford Jill 
Published: January 2023

Abstract
The Triggering Receptors Expressed on Myeloid cells (TREM) gene cluster contains receptors that regulate a variety of leukocytes. TREM1 synergizes with Toll Like Receptors to promote cytokine production in neutrophils, macrophages and dendritic cells (DC). Paradoxically, although TREM1 and TREM2 both signal via DAP12, TREM2 is a negative regulator of inflammation in macrophages and DC. TREM2 is expressed in the colon but its role in colitis and colitis-associated cancer (CAC) is poorly understood. We found TREM2 expressed in monocytic myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages (TAM). Moreover, TAM expressed a apparent TREM2 ligand. To test the role of TREM2 in inflammation-induced cancer we studied the azoxymethane (AOM)-dextran sulfate sodium (DSS)-induced model of CAC. DSS administration led a pronounced influx of myeloid cells and upregulation of both TREM1 and TREM2. Surprisingly, we found that Trem2-/- mice had less severe colitis as indicated by reduced histological scores, colon shortening and inflammatory cytokine production in the colon. AOM-DSS carcinogenesis in Trem2-/- mice resulted in slightly reduced total polyp counts and mitotic epithelial cells but a substantial reduction in the numbers of advanced carcinomas compared to wild type mice. Taken together, our data suggest an important role for TREM2 in the regulation of colitis and CAC likely via control of epithelial proliferation during colonic injury and inflammation.
Activation of IL-6/STAT3-signaling cascade induces suppression of antigen presentation by human dendritic cells (TUM4P.917)
Authors: Kitamura Hidemitsu , Ohno Yousuke , Ohtake Junya , Kaneumi Shun , Kishikawa Takuto , Terada Satoshi , Sumida Kentaro , Masuko Kazutaka , Kita Toshiyuki , Takahashi Norihiko , Taketomi Akinobu 
Published: January 2023

Abstract
Immunosuppression in tumor microenvironments is one of the critical issues for cancer immunotherapy. We have been demonstrated that IL-6, a pleiotropic cytokine that regulate the growth, differentiation and survival of a variety of cells, significantly inhibited maturation of murine dendritic cells (DCs) via STAT3 activation both in vitro and in vivo. In this study, we focused on IL-6/STAT3-signaling cascade in human dendritic cells (DCs), and investigated the effects of IL-6 on antigen-presenting ability of DCs. Surface HLA class I, HLA class II, and costimulatory molecules of human DCs were evaluated after the stimulation with IL-6 in vitro. As a result, HLA-DR and CD86 expressions were significantly reduced by IL-6 treatment of human DCs. The reduction was remarkably blocked in the presence of STAT3 inhibitor. In addition, IFN-γ production by T cells after TCR-stimulation was significantly suppressed in the presence of IL-6-treated DCs compared with control DCs. Moreover, we found that activation of antigen-specific CD4+ T cells by DCs was remarkably reduced by the IL-6-treatment. In this study, we demonstrated that IL-6/STAT3-signaling cascade was one of the regulating factors for antigen presentation of DCs. From these findings suggest that IL-6/STAT3 signaling cascade regulates immunosuppressive function of human DCs, which would be a promising target for improving the effects of cancer immunotherapy.
IRF-8 levels constrain aberrant myelopoiesis and improve immune control in the metastatic tumor microenvironment (TUM4P.901)
Authors: Netherby Colleen , Waight Jeremy , Miller Austin , Bogner Paul , Liu Kebin , Abrams Scott 
Published: January 2023

Abstract
Abnormal myelopoiesis is common in patients with diverse malignancies, including breast cancer. The myeloid compartment is essential for the induction of host immunity, thus alterations in myelopoiesis may help explain the failure to control neoplastic disease. Therefore, understanding the molecular basis of myeloid dysfunction may improve the efficacy of cancer immunotherapies. Given that interferon regulatory factor-8 (IRF-8) is crucial for myeloid differentiation, we hypothesized that changes in this master regulator underlie a novel mechanism for the altered myelopoiesis and myeloproliferative phenotypes observed in cancer, such as the accumulation of myeloid-derived suppressor cells (MDSC). We recently showed that tumors downregulate IRF-8 expression in myeloid progenitors in a cytokine-driven STAT3 or STAT5-dependent manner, resulting in MDSC generation. This suggested that IRF-8 expression functions as a ‘rheostat’ which directs the fate of the tumor-induced myeloid response. To test this, we examined the impact of modulating IRF-8 levels in two mouse mammary tumor models of lung metastasis using a newly designed genetic gain-of-function approach. In both models, we observed a significant reduction in lung metastases and the frequency of MDSC in IRF-8 transgenic mice compared to wild-type controls. Altogether, our results reveal an unrecognized role for IRF-8 in the host response against metastasis, which may lead to new therapeutic targets in cancer immunotherapy.
Genomic ablation of HMGB1 in dendritic cells mitigates their capacity to both promote carcinogenesis and effectively confer anti-tumor immunity as vaccines (TUM4P.919)
Authors: Vernon Philip , Liang Xiaoyan , Wang Wenqian , Schapiro Nicole , Lowe Devin , Billiar Timothy , Zeh Herbert , Lotze Michael 
Published: January 2023

Abstract
HMGB1 is the prototypical damage associated molecular pattern (DAMP) molecule. It plays a critical role in many aspects of DC biology. To study the role of DC HMGB1 in tumor immunity, DC-specific HMGB1-/- mice (DCH) were generated. In both subcutaneous and liver metastasis models of pancreatic and colorectal cancer respectively, DCH mice exhibited inhibited tumor growth. DC HMGB1 is also critical for carcinogenesis; bone marrow from DCH mice was transplanted into mice expressing pancreas-specific oncogenic Kras (KC). KC mice given DCH bone marrow exhibited lower frequencies of malignant pancreatic and less pro-tumor inflammatory infiltrate. Genomic loss of HMGB1 also negatively impacted other aspects of DC functionality. Mice inoculated subcutaneously with pancreatic and melanoma cancer cell lines and then vaccinated with bone-marrow derived KO DCs pulsed with tumor antigen were unable to achieve therapeutic protection in both models. Although both WT and KO DCs were able to stimulate allogeneic lymphocyte proliferation, KO DCs skewed responses away from Th1/Th2 phenotypes, and favored Treg/Th17 polarization. KO DCs also expressed lower levels of both RAGE and TLR4, suggesting these signaling pathways may be impaired in DCH mice, thus accounting for our observations.
Negative impact of myeloid-derived suppressor cells on CD8 effector T cell trafficking within the tumor microenvironment (TUM4P.924)
Authors: Ku Amy , Muhitch Jason , Abrams Scott , Evans Sharon 
Published: January 2023

Abstract
The success of immunotherapy and, unexpectedly, chemotherapy and radiation hinges on cytotoxic T cells gaining access to tumor targets. These observations have prompted interest in developing strategies to improve T cell trafficking in tumors. Here, we report that the ability of tumor vessels to respond to IL-6-dependent preconditioning regimens that boost CD8 effector T cell homing is temporally and inversely related to the expansion of myeloid-derived suppressor cells (MDSC) within the tumor microenvironment. Using real-time intravital imaging and immunofluorescence histology, IL-6 therapies were shown to convert vessels from T cell-low to -high recruitment sites in murine tumors with minimal MDSC infiltration (i.e., CT26 colorectal, B16 melanoma, EMT6 mammary tumors). This conversion requires induction of the ICAM-1 trafficking molecule on tumor vessels. Conversely, mammary (4T1, AT-3 and PyMT-MMTV) and pancreatic (Pan02) tumors with high MDSC burdens were refractory to IL-6 therapies, but became responsive after acute MDSC depletion. To further investigate contributions of MDSC to poor trafficking, IL-6-responsive tumors were admixed at a ratio of 2:1 with MDSC from tumor-bearing mice. Sustained elevation of MDSC in admixed tumors resulted in failure to support increased T cell trafficking in response to IL-6-dependent therapy. Taken together, these findings identify a novel role for MDSC in immune evasion by limiting T cell trafficking at tumor vascular loci.
Neutrophil recruitment into premalignant lesions differentially influences tumor growth versus progression. (TUM4P.928)
Authors: Blaisdell Adam , Daikoku Takiko , Tay Chin Siean , Dey Sudhansu , Erlebacher Adrian 
Published: January 2023

Abstract
Neutrophils possess a diverse arsenal of enzymatic and chemical weapons that can inflict damage upon pathogens and host cells alike. Most tumors harbor neutrophils even from the earliest stages of progression (premalignancy), wherein these weapons could, in theory, either kill a tumor cell or enhance its malignant potential via mutagenesis. However, the interaction between neutrophils and premalignant tumor cells has not been well characterized. To this end, we utilized a mouse model of endometrial carcinoma with defined stages of premalignancy and malignancy that faithfully represent human endometrial cancer progression. We observed robust neutrophil infiltration within premalignant lesions, accompanied by elevated expression of G-CSF, Il1a, Il1b and CXCR2 ligands. Blockade of G-CSF signaling substantially reduced tumor-associated neutrophil numbers and had profound and contrasting effects on tumor growth versus progression. Neutrophil-depleted lesions were larger and contained an increased number of proliferating tumor cells, but concomitantly showed less dysplasia and a reduced amount of DNA damage within tumor cells. Additionally, neutrophil recruitment into premalignant lesions relied heavily upon IL-1 signaling to induce the expression of G-CSF and CXCR2 ligands. These data establish a double-edged sword paradigm for neutrophils in the premalignant tumor microenvironment as agents of both tumor cell death and malignant potentiation.
Produce immunogenic monocyte derived-dendritic cells by Toxoplasma gondii proteins (TUM4P.904)
Authors: Boghozian Roobina , Mirzaei Reza , Hadjati Jamshid 
Published: January 2023

Abstract
Background: DCs based immunotherapy is considered as a promising approach for activating immune responses against tumor antigens. It has been believed that TLR ligands derived from microorganisms are more potent in activating DCs. In this study, the effects of Toxoplasma gondii (T.gondii) protein and DNA components on human DCs maturation were evaluated. Method: T.gondii Tachyzoites were obtained by intraperitoneal passage in mice. Protein and DNA components were extracted by relevant kits. Subsequently, protein lysate was fractionated by increasing amounts of ammonium sulfate and after that by ion-exchange chromatography. Protein fractions and DNA components were added to human monocyte derived DCs. After 20h, phenotype markers (CD80, CD83, CD86 and HLA-DR) on DCs were determined by flow cytometry. Also, IL-12p70 and IL-10 concentration in DCs supernatants were evaluated. Ultimately, the effects of different fractions on phagocytosis activity of DCs were compared. Results: We found that T.gondii protein components are more potent in activating DCs than DNA components. Moreover, there is a considerable difference in phenotype markers and phagocytic activity between DCs matured by different protein fractions. Also, the ability of some protein fractions is higher in activating DCs to produce IL-12p70 and lower IL-10. Conclusion: The protein fractions with high ability can be used to produce potent DCs for instructing immune responses against tumor antigens.
Effect of blocking different TGF-β isoforms on tumor immunity (TUM5P.937)
Authors: Terabe Masaki , Robertson Faith , Mizra Amer , Berzofsky Jay 
Published: January 2023

Abstract
TGF-β is a pleiotropic cytokine with three isoforms that have significantly different physiological functions. This cytokine has potent immunosuppressive activity and has been shown to play critical roles in tumor development. TGF-β1 is also necessary for the development of T cell subsets as well as MDSC. We have previously reported that blockade of TGF-β by a monoclonal antibody 1D11, which neutralizes all three isoforms of TGF-β, improves immunosurveillance and vaccine efficacy in mouse tumor models. However, the role of individual isoforms of TGF-β is unknown. In this study, we examined the role of different TGF-β isoforms in tumor immunity by using anti-TGF-β with unique specificities in two tumor models, a lung metastasis of the BALB/c CT26 colon carcinoma and a subcutaneous TC1 lung epithelial cell tumor in B6. In the CT26 model, significant reduction of tumor burden was seen through the dual blockade of TGF-β1 and 2, or inhibition of all three TGF-β isoforms. In the TC1 model, the mice were treated with a peptide vaccine when the tumor size reached at least 5mm in diameter with/without anti-TGF-β. Although the vaccine alone induced significant regression of tumors, both types of anti-TGF-β facilitated the vaccine efficacy. These results suggest that blockade of only TGF-β1 and 2 is sufficient to facilitate natural immunosurveillance or enhance vaccine-induced tumor immunity. Currently we are investigating the effect of TGF-β isoform blockade on T cell subsets.
Antibodies targeting the Rhizopus oryzae invasin, CotH3, enhance neutrophil killing (VAC6P.952)
Authors: Lin Lin , Gebremariam Teclegiorgis , Ibrahim Ashraf 
Published: January 2023

Abstract
Mucormycosis is a lethal fungal infection that afflicts the immunocompromised host such as those suffering from hyperglycemia, ketoacidosis, other forms of acidosis, neutropenia, and those receiving corticosteroids. Rhizopus spp. are the most common pathogens isolated from patients with mucormycosis. Previously, we identified R. oryzae CotH3 as a surface protein which mediates fungal invasion of host cells during mucormycosis. We found rabbit polyclonal anti-CotH3 antibodies to significantly block host cell invasion by R. oryzae and protect mice from R. oryzae infection. However, it is unclear if CotH3 antibodies enhance phagocyte killing of the fungus. Here, we used flow cytometry and mouse neutrophil killing assays to determine the binding and opsonophagocytic killing capacities of varying concentrations of anti-CotH3 antibodies against R. oryzae, respectively. Anti-CotH3 antibodies showed high-affinity binding (&gt;99%) to R. oryzae even when low concentration of the antibodies were used (e.g. 3mcg dose). CotH3 antibodies also widely bound to other Mucorales known to cause mucormycosis including Mucor circinelloides, Lichtheimia, Rhizomucor, and Cunninghamella. Neutrophil kill assay has shown Anti-CotH3 antibodies to dramatically enhance mouse neutrophil killing by &gt; 2-fold increase vs. control with preimmune antibodies (P&lt;0.04). Our studies warrant the further development of monoclonal antibodies targeting CotH3 protein as a novel therapy against mucormycosis.
Approaches for sampling human antibodies for repertoire analysis and reagent development (VAC6P.953)
Authors: Chen Zhifeng , Kannan Harilakshmi , Derstine Lauren , Qiu Ping , Prokop Michael , Touch Sinoeun , Su Bin , Dubey Sheri , Wang I-Ming , ter Meulen Jan , DiStefano Daniel , Vora Kalpit 
Published: January 2023

Abstract
Purpose: Majority of vaccines rely on Human humoral response for protection. Sampling antibody response is useful for understanding protection correlates of vaccine and isolating reagent antibodies for lifecycle management. We used 3 approaches delineated below to isolate Rubella neutralizing antibodies from plasmablast B-cells separated from human subjects recently vaccinated with MMRII vaccine. Direct B cell cloning: Plasmablast B cells isolated from donors at day 7 after vaccination were used directly as templates in the RT-PCR amplification of antibody genes. Antigen-specific Fabs were isolated following molecular cloning and ELISA screening. Deep Sequencing: RNA extracted from plasmablast B cells was used as template in RT-PCR amplification. The PCR products of antibody genes were subjected to deep sequencing, retrieved antibody VH, VL and VK sequences were rank ordered based on the frequency and, top antibody sequences in each category were expressed as Fabs and tested. Immune Library Construction/Phage Display: Antibody heavy and light chain variable regions from plasmablast B cells were cloned into bacterial phage display vectors to make immune libraries, specific Fabs were identified following bio-panning. Result: All together 25 specific antibodies against Rubella were isolated through the above approaches, the best Fab ELISA binder had an EC50 of 3.5ng/mL (0.07nM). Two of the Fabs demonstrated neutralizing activity with EC50’s of 1µg/mL (20nM) and 4µg/mL (80nM).
Long-lived bone marrow plasma cells are inhibited by protein A in Staphylococcus aureus infection. (VAC6P.955)
Authors: Keener Amanda , Thurlow Lance , Richardson Anthony , Vilen Barbara 
Published: January 2023

Abstract
Long-lived plasma cells are now recognized as major contributors to protection against bacterial infection through their constitutive secretion of antibody. Staphylococcus aureus is a growing concern due to expansion of antibiotic resistance and causes recurring infections without inducing long-lived immunity. Little is known about the nature and longevity of the adaptive response to S. aureus. Protein A on the surface of S. aureus promotes virulence and inflammation which can alter B cell function and hinder long-lived plasma cell development. Although similar B cell responses were induced by WT and protein A-deficient infection and secondary challenge, long-term antigen-specific antibody levels waned after challenge with WT S. aureus, but were maintained after challenge with the protein A-deficient mutant. This suggests that long-lived B cell immunity is disrupted by protein A. We developed an ELISpot assay to detect S. aureus-specific antibody secreting cells and found that they accumulated in the bone marrow niche after protein A-deficient, but not WT challenge. The absence of bone marrow plasma cells after WT challenge was not a result of defects in germinal center or memory B cell formation. Our work indicates that protein A interferes with migration and/or survival of long-lived bone marrow plasma cells after S. aureus infection. It also provides insight into the requirements for successful development of long-lived immunity to bacterial pathogens.