bluuebunny/crossref_metadata_2025_split huggingface dataset:
A 13-color parametric analysis for TBNK subset for the CytoFLEX instrument (TECH3P.935)
Authors: Lawson Jonel , Fischer Karen , Tung James 
Published: January 2023

Abstract
CytoFLEX is the newest cytometer offered by Beckman Coulter, Inc. that provides up to 3 lasers, 13 colors for detection in a compact system. This instrument incorporates the latest technology, Fiber array photodiode, FADP, to provide high resolution, dynamic range and sensitivity in signal detection. To demonstrate the ability of its multicolor detection, the standard multicolor panel design principles were followed to develop a 14 marker, 13 color experiment. The cocktail design process also took into account the desired immunophenotypic markers, the available fluorochromes, and the use of well-defined control populations to identify the classic TBNK populations. Data was collected from normal, whole blood stained with single color reagents and lysed with VersaLyseTM Lysing Solution. Data was analyzed using the CytExpert software and is presented here with special emphasis on the optimization of flow analysis and compensation visualization. Results show that all TBNK populations were well resolved, notably the CD25+ subset in CD4+ T-cells, even though this maker was placed in the PC7 channel, which normally has poor resolution. The data presented demonstrates the ability of the CytoFLEX to provide sound detection and resolution for even the most dim versus negative population.
Anti-inflammatory and neuroprotective effects of GILZ analogs. (THER7P.951)
Authors: Srinivasan Mythily , Chopra Nipun , Bayon Baindu , Lahiri Debomoy 
Published: January 2023

Abstract
Increased activation of the p65 subunit of nuclear factor kappa-B (NF-κB) upregulate inflammatory and apoptotic factors that mediate neuroinflammation and neurodegeneration in chronic neurological diseases, suggesting that activated p65 is an excellent therapeutic target. Glucocorticoid induced leucine zipper (GILZ) is a p65 interacting protein, that sequesters activated p65 in the cytoplasm and suppresses NF-κB. The GILZ:p65 interaction is localized to the proline rich region at the carboxy terminus of GILZ. Previously we reported that a synthetic peptide (GILZ-P) derived from the p65 binding domain of GILZ ameliorated experimental encephalomyelitis. To determine the potential application of GILZ-P in humans, we evaluated the toxicity and efficacy of the peptide drug in mature human neuroblastoma cells, macrophage like THP-1 cells and fetal neuronal cells. Treatment with GILZ-P at a wide range of concentrations commonly used for peptide drugs was non-toxic as determined by the cell viability and apoptosis assays in THP-1 cells. Functionally GILZ-P suppressed pro-inflammatory cytokine secretion by proteolipid protein or amyloid-β (Aβ1-42) stimulated mature neuroblastoma cells and suppressed glutamate secretion by activated macrophages by inhibiting nuclear translocation of p65. Collectively, GILZ-P exhibits therapeutic potential for chronic diseases wherein persistent inflammation precipitates neurodegeneration such as progressive multiple sclerosis and Alzheimer’s disease.
Arctigenin regulates Th1 and Th17 differentiation and ameliorates experimental autoimmune encephalomyelitis (THER7P.958)
Authors: Zhang Rongxin , Li Wen , Zhang Zhihui , Zhang Zimu , Xue Zhenyi , Li Yan , Zhang Lijuan , Da Yurong , Yao Zhi 
Published: January 2023

Abstract
Arctigenin is a phenylpropanoid dibenzylbutyrolactone lignan extracted from Arctium lappa (L.), and exhibit antitumor, antioxidant and neuron protective activities. But the effects of arctigenin on CNS inflammatory diseases are still unclear. Experimental autoimmune encephalomyelitis (EAE) is an animal model thought to be a T cell-mediated inflammatory demyelination disease. In this study, we in vivo explore the effect of arctigenin on mouse CD4+ effect T helper cell functions. Furthermore, in vitro explore the therapeutic effects and cellular mechanisms of arctigenin on EAE. Arctigenin treated mice are resistant to EAE, the clinical score of arctigenin treated mice is significantly reduced. Histochemistry assay of spinal cord sections also showed that arctigenin reduced inflammation and demyelination in EAE mice. The Th1 and Th17 cells were decreased in the spleens and lymph nodes of arctigenin treated EAE mice, while Th1 cells in CNS did not have significant changes. The expression of Th1 and Th17 cell proliferation and polarizing cytokines are inhibited in arctigenin treated mice spleens and lymph nodes. The Th1 cytokine IFN-γ and transcription factor T-bet, and Th17 cytokines IL-17A, IL-17F, and transcription factor ROR-γt, were significatly suppressed in vitro and in vivo. These suggest that arctigenin have properties of anti-inflammation and immunosuppression, which could be a potential therapeutic drug for multiple sclerosis and other autoimmune inflammatory diseases.
TNF regulated disability in experimental autoimmune encephalitis (THER7P.946)
Authors: Valentin-Torres Alice , Phares Timothy , Stohlman Stephen 
Published: January 2023

Abstract
In EAE, abrogation of IFN-γ signaling in astrocytes results in a progressive clinical disease, with increased CNS inflammation and demyelination, making this an excellent mouse model to study progressive multiple sclerosis (MS). Microglia and infiltrating macrophage TNF production is elevated during progressive EAE which is linked to pathology of several neurodegenerative disorders including MS. Moreover, anti-TNF therapies have been successfully used to treat several autoimmune diseases such as rheumatoid arthritis, Crohn’s disease, and psoriatic arthritis. Neutralization of both soluble and membrane bound TNF during progressive EAE significantly ameliorated disease progression and decreased demyelination. TNF blockade reduced the overall infiltration of immune cells into the CNS including IL-17 and IFN-γ producing T cells by restoring the blood brain barrier integrity. A reduction in the number of microglia and macrophages expressing MHC II+ correlated with decreased inflammation. Conversely, anti-TNF therapy increased IL-10 and IL-27 within the CNS without altering Foxp3+ Treg frequencies. Overall, these data demonstrate that TNF is an important mediator of the disability, inflammation, and pathology during progressive EAE. Our results demonstrate that TNF is a potential candidate to treat progressive forms of MS.
CD4+CD25+Foxp3+ cells from JNK-/- mice prolong pancreatic allograft survival in type 1 diabetic mice (TRAN2P.970)
Authors: Tripathi Deepak , Venkatasubramanian Sambasivan , Cheekatla Satyanarayana , Paidipally Padmaja , Welch Elwyn , Tvinnereim Amy , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
In the current study we asked whether modified CD4+CD25+FoxP3+ regulatory T-cells (T-regs) can induce long term allograft tolerance. Pancreatic islets from BALB/c mice survived in a type 1 diabetic C57BL/6 mice (recipient mice) liver up to 18-21 days (n=5). Preloading islets with T-regs from WT (wild type) C57BL/6 mice, enhanced survival up to 37 days (N=5 and p = 0.003). Further preloading islets with T-regs from JNK1-/- mice (C57BL/6 background) enhanced survival up to 107 days (N=5 and p = 0.002). In contrast preloading islets with T-regs from FAS-/- (C57BL/6 background) did not enhance islet survival beyond 45 days. T-cell receptor stimulation of T-regs from JNK1-/- mice produced significant high amount of IL-10 (14.67 ± 2.784 vs 3.803 ± 0.8325, p = 0.009) and TGF-β (27.93 ± 7.563 vs 9.920 ± 1.212, p = 0.05) compared to T-regs from WT mice. Preloading islets with T-regs from JNK1-/- mice reduced IL-1β (30.91 ± 2.41 vs 16.13 ± 2.76; p = 0.003), TNF-α (3.802 ± 0.69 vs 1.888 ± 0.26; p = 0.03) IL-17 (24.82 ± 4.07 vs 4.708 ± 1.57; p = 0.0017). Studies are underway to determine the JNK1-/-T-regs mediated mechanisms involved in prolongation of allograft survival.
Deficiency of indoleamine 2,3-dioxygenase in donor bone marrow cells leads to exacerbation of graft-versus-host disease through enhanced generation of neutrophils and effector T cells (TRAN2P.965)
Authors: Ju Ji Min , Choi Eun 
Published: January 2023

Abstract
Indoleamine 2,3-dioxygenase-1 (IDO-1) is a tryptophan metabolic enzyme expressed by various types of cells including dendritic cells, myeloid-derived suppressor cells, and intestinal epithelial cells. The enzyme is also well known for immune suppressive function, while the underlying mechanism is still obscure. To understand cellular mechanism underlying the suppressive role of IDO-1, especially expressed by donor-derived myeloid cells in generation of graft-versus-host disease (GVHD), we transplanted T cell-depleted (TCD) BM cells from Ido-1-/- mice together with wild type (WT) T cells on C57BL/6 (B6) background into MHC-matched allogeneic BALB.B mice. Compared with BALB.B hosts that received WT TCD-BM and T cells, those of Ido-1-/- TCD-BM and WT T cells showed exacerbated GVHD. T cell frequencies in blood and target organs, and their productions of effector cytokines (IFN-γ- and IL-17) were enhanced in the recipients of Ido-1-/- TCD-BM, compared with those of WT TCD-BM. Generation of CD11b+Gr-1+ cells, especially of Ly6GhiLy6lo cells, was also enhanced in the formers. The CD11b+Gr-1+ cells originated from Ido-1-/- TCD-BM exerted lower suppressive activity over proliferation and cytokine production of co-incubated allo-reactive T cells. Proportions of CD244- neutrophils within the Ly6GhiLy6lo population of Ido-1-/- TCD-BM origin were significantly higher than that of WT TCD-BM. These results demonstrate implication of IDO-1 function in neutrophil generation.
Liver NK1.1 cells and IL-22 promote pancreatic islets allograft survival in type 1 diabetic mice (TRAN2P.969)
Authors: Tripathi Deepak , Venkatasubramanian Sambasivan , Cheekatla Satyanarayana , Paidipally Padmaja , Welch Elwyn , Tvinnereim Amy , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
We determined the role of liver NK1.1 cells in allograft survival. Pancreatic islets from BALB/c mice survived in a type 1 diabetic C57BL/6 mice liver up to 15-21 days (n=5, p=0.001). Anti NK1.1 but not isotype control antibody treatment of type 1 diabetic C57BL/6 mice before and after pancreatic islet transfer rejected graft within 3 days (n=5, p=0.001). Anti-NK1.1 antibody treatment enhanced IL-1β (45.13 ± 5.9 vs 94.25 ± 5.5; p = 0.0003), TNF-α (4.746 ± 2.4 vs 11.92 ± 1.8; p = 0.047) and IFN-γ (2.714 ± 0.45 vs 4.344 ± 0.4; p = 0.036) and inhibited IL-22 (24.82 ± 4.07 vs 11.01 ± 2.186; p = 0.01) gene expression by liver lymphocytes of recipient mice. NK1.1 cells are major source of IL-22 in islets recipient mice liver and anti-IL-22 antibody treatment of recipient mice reduced allograft survival to 6-9 days compared to 18-21 days in isotype control antibody treated mice. Recombinant IL-22 but not IFN-γ enhanced insulin production by pancreatic islets of BALB/c mice (102.0 ± 22.65 vs 33.41 ± 12.07; p = 0.03) and enhanced expression of genes Reg2, Reg3a and Reg3g involved in islet survival. Further studies indicated that immunization and challenge of type 1 diabetic C57BL/6 mice with TLR9 agonist can enhance IL-22 production by NK1.1 cells and prolongs pancreatic allograft survival from 15 days to 45 days. The sum of our studies suggests that IL-22 produced by liver NK1.1 cells enhance allograft survival and TLR9 agonist can be used to prolong pancreatic allograft survival.
Expression of indoleamine 2.3-dioxygenase (IDO) by tumors induces local and systemic immunosuppressive effects in a murine melanoma model (TUM10P.1029)
Authors: Holmgaard Rikke , Zamarin Dmitriy , Munn David , Merghoub Taha , Wolchok Jedd 
Published: January 2023

Abstract
IDO is found in the majority of human tumors and has been described as an important contributor to the development of an immunosuppressive tumor microenvironment that blocks the action of cytotoxic antitumor effector T cells. In order to delineate the mechanisms of IDO-induced immunosuppression in melanoma, we generated B16 melanoma cell lines overexpressing IDO (B16-IDO). When implanted into mice, B16-IDO tumors exhibited aggressive tumor growth, characterized by more rapid tumor progression and local invasion. Furthermore, constitutive expression of IDO by B16 cells conferred resistance to T-cell-targeting immunotherapies, such as CTLA-4 and PD-1 blocking antibodies. Interestingly, the immunosuppressive effect of IDO overexpression was evident not just within the tumor, but systemically as well. Analysis of the tumors in vivo, showed that this effect was accompanied by changes favoring a highly inhibitory tumor microenvironment characterized by significant reduction on local tryptophan concentrations and lack of effector T cells at the tumor site. Most strikingly, we observed that immunosuppressive cell populations such as regulatory T cells and myeloid-derived suppressor cells dominated the cellular infiltrates of B16-IDO tumors. All together our findings delineates IDO expression as an important mechanism driving the institution of an inhibitory tumor microenvironment and tumor progression, and provides a strong rationale for therapeutic targeting of this pathway.
IL-22 induces colon cancer proliferation through STAT3 and PRC2 signaling (TUM10P.1036)
Authors: Sun Danfeng , Lin Yanwei , Kryczek Ilona , Zou Weiping 
Published: January 2023

Abstract
Background: IL-22(interleukin-22) is a newly identified cytokine which has been recently highlighted owing to its biological significance in carcinogenesis. However, the role of IL-22 in colon cancer proliferation has remained unclear. Since both JAK-STAT signaling and Polycomb Repression complex (PRC) 2 play crucial roles in epithelial proliferation. We investigated the mechanism by which IL-22 induces colon cancer proliferation. Methods: Colon cancer cells proliferation was detected by FACS and H3 Thymidine Incorporation. Cell cycle related genes were tested by Real-Time PCR. Lentiviral vector encoding gene specific shRNAs (STAT3, EZH2, SUZ12) and scramble particles were used to transfect colon cancer cells. ChIP assays were performed to detect the binding of different proteins and DNA. Results: IL-22 significantly induces colon cancer cells proliferation through STAT3 signaling. This increased proliferation involved the inhibition of p16 and p18 through aberrant promoter histone methylation H3K27me3. PRC2 catalyzes promoter H3K27me3 of p16 and p18, which can be up-regulated by IL-22. Furthermore, STAT3 can bind to PRC2 members (SUZ12 and EED), which also can be up-regulated by IL-22 stimulation. Conclusion: IL-22 can induce human colon cancer cells proliferation by activation of STAT3, which was phosphorylated and directed bind to the PRC2 complex, especially SUZ12 and EED, and consequently catalyzed the H3K27m3 of cell cycle check-point genes p16 and p18.
Induction of tissue-resident TFH-like cells in human cancer links monocyte activation to M2b macrophage polarization (TUM10P.1021)
Authors: Chen Min-Min , Kuang Dong-Ming 
Published: January 2023

Abstract
The existence, regulation, and functions of IL-21+ immune cells are poorly defined in human cancers. Here we identified a subset of protumorigenic IL-21+ TFH-like cells in human hepatocellular carcinoma. These cells were the major source of IL-21 in tumors and represented about 10% of the CD4+ T cell population at levels comparable to amounts of TFH cells present in lymph nodes. However, the TFH-like cells displayed a unique CXCR5−PD-1−BTLA−CD69high tissue-resident phenotype with substantial IFN-γ production, which differed from the phenotype of TFH cells. TLR4-elicited innate monocyte inflammation was essential for IL-21+ TFH-like cell induction in tumors, and activation of STAT1 and STAT3 was critical for TFH-like cell polarization in this process. Importantly, the TFH-like cells operated in IL-21-IFN-γ-dependent pathways to induce plasma cell differentiation and thereby create conditions for M2b macrophage polarization. Thus induction of TFH-like cells links innate inflammation to immune privilege in tumors.
Early stage human lung tumors give rise to a subpopulation of tumor-associated neutrophils with the composite characteristics of neutrophils and antigen-presenting cells (TUM10P.1042)
Authors: Eruslanov Evgeniy , Bhojnagarwala Pratik , Quatromoni Jon , Moon Edmund , Stephen Tom , Akimova Tatiana , Hancock Wayne , Conejo-Garcia Jose , Feldman Michael , Albelda Steven , Singhal Sunil 
Published: January 2023

Abstract
Tumor-associated neutrophils (TANs) represent a significant portion of inflammatory cells in the lung tumor; however, the role of TANs in cancer progression remains largely unexplored in humans. Given the key role of cytotoxic T lymphocytes in mediating anti-tumor immunity, the goal of this study was to identify subsets of TANs that might have specific roles in regulation of T cell responses in patients with lung cancer. Our analysis of 37 early-stage human lung tumors revealed two major sub-populations of TANs. One subset of “canonical” TANs expressed classic neutrophil markers. A second subset of “hybrid” TANs displayed a combination of neutrophil (Arg1+MPO+CD66b+CD15+) and antigen-presenting cell (CD14+HLA-DR+CCR7+CD86+) markers. These hybrid TANs comprised 0.5-25% of all TANs. Using human bone marrow (BM) cells, we identified conditions in which the immature granulocytes could be differentiated into cells that highly resemble hybrid TANs. Functionally, the BM-derived hybrid subset and hybrid TAN were able to: 1) stimulate antigen non-specific T cell proliferation, 2) induce the proliferation of allogeneic T cells, 3) trigger antigen-specific memory T cell responses, and 4) cross-present the NY-ESO peptide to TCR-transfected CD8 cells. Together, our results reveal a specialized subpopulation of TAN capable of triggering and supporting T cell responses. This property of hybrid neutrophils may provide new opportunities to boost the efficacy of current immunotherapy.
Defective TRAF3 modulates alternative NF-kB signaling and cytokine expression to promote cancer cell survival in HPV positive head and neck cancer (TUM10P.1049)
Authors: Zhang Jialing , Chen Hui , Yang Xinping , Guven Emine , Nussinov Ruth , Chen Zhong , VanWaes Carter 
Published: January 2023

Abstract
Persistent infection of high-risk human papillomaviruses (HPV) is a known pathogen associated with head and neck squamous cell carcinoma (HNSCC), especially in tonsil, oropharynx and oral cavity. The incidence of HPV+ HNSCC has been increased recently in the Unites States, while the molecular mechanism of virus promoting HNSCC has not been fully understood. Recently, The Cancer Genome Atlas (TCGA) project revealed unique homozygous and heterozygous deletions and mutations of TNF receptor-associated factor 3 (TRAF3) in HPV associated HNSCC tumors. Low expression of TRAF3 was observed in HPV positive HNSCC samples and cell lines with TRAF3 deletions. These tumors had elevated expression of NF-kB subunits involved in alternative pathway, such as NF-kB2 and RelB, often associated with increased NIK, cIAP1 and cIAP2 expression. Overexpression of TRAF3 in HPV+ cell lines significantly suppressed alternative NF-kB subunit NF-kB2/RELB nuclear localization, while increasing expression of IFNα/β and IL10 implicated in anti-viral immunity. Further, overexpression of TRAF3 inhibited cell growth, colony formation, and migration, and sensitized cells to TNF-a and cisplatin induced cell death. Our data revealed that defective TRAF3 activated alternative NF-kB signaling and suppressed anti-viral cytokine expression, which promoted cancer cell survival and drug resistance, especially in HPV+ HNSCC.
Inflammatory chemokine CCL3 Leads to enhanced antitumor development in the draining lymph node (TUM10P.1053)
Authors: Allen Frederick , Bobanga Iuliana , Barkauskas Deborah , Myers Jay , Huang Alex 
Published: January 2023

Abstract
CCL3 has been shown to be important in orchestrating cellular contacts in vaccinated lymph nodes. Moreover, dendritic cells can mediate both tumor tolerance and anti-tumor immunity in some tumor models. We hypothesize that incorporating CCL3 into tumor vaccines can enhance anti-tumor immunity. We inoculated naïve recipient mice with either WT or CCL3-secreting colon tumors. CCL3-secreting tumors showed enhanced antitumor immunity and suppressed growth kinetics in immunocompetent mice versus WT-tumors. Relative to non-injected controls or WT-injected tumors, 3 days after injections CCL3-secreting tumors revealed an increase of CD4+, CD8+, and CD19+ lymphocyte in the draining lymph node. Concurrently we conducted in vitro BMDCs experiments to assess the effects the tumor microenvironment (+/- CCL3) has on the capacity of DCs to upregulate T cell activation and adhesion molecules. BMDCs differentiated in the presence of CCL3 showed a greater capacity to upregulation CD86, ICAM1, CD11b, and CCR7 before and after LPS stimulation. Moreover, BMDCs cultured in the presence of WT and CCL3-secreting tumor supernatants revealed an increase in CD86, MHCII, and CCR7 before and after LPS stimulation. Together these data reveal that CCL3 in the tumor microenvironments leads to a reduction in tumor growth and early lymphocyte accumulation in the DLN, which may be a result of an enhanced capacity for DCs to home to the DLN and activate antitumor T cells.
Attenuated rabies virus promotes CNS immune infiltration and tumor necrosis in a mouse model of glioma (TUM10P.1059)
Authors: Bongiorno Emily , Hooper Douglas 
Published: January 2023

Abstract
Malignant glioma is resistant to conventional treatment regimens, and recent immunotherapeutic strategies have had limited success, presumably due to lack of immune cell infiltration and function in the brain. Attenuated rabies virus (RABV) is unique in its ability to allow the therapeutic infiltration of immune cells into the CNS, hence we speculated that infection might have a beneficial effect in the GL261 mouse model of glioma. We found that infection of mice with attenuated RABV after the establishment of a GL261 CNS tumor significantly prolongs their survival. Improved survival is associated with the appearance of tumor necrosis despite the inability of the virus to replicate in the tumor. Infected tumor bearing animals also have higher levels of GL261-specific IgG2A antibody, indicating that the Th1 arm of the response has been promoted. Increased B, T and NK cell infiltration as well as elevated IFNγ and ICAM expression is also observed in CNS tissues of infected mice. In addition tumor-associated cells bearing PD-L1, FasL, CD133, and M2 markers are strikingly less abundant in the infected mice compared to uninfected controls. Our results suggest that the tumor necrosis caused by infection with RABV is the result of altered cell activity within the tumor rather than a direct effect of the virus. Increased effector presence in the CNS combined with fewer regulatory and evasive tumor-associated cells contribute to better tumor killing in RABV-infected mice.
Exosomes released from Doxorubicin-treated breast tumor promote lung metastasis through miR126 mediated induction of MDSCs (TUM10P.1060)
Authors: Deng Zhongbin , Teng Yun , Rong Yuan , Zhuang Xiaoying , Mu Jingyao , Zhang Lifeng , Samykutty Abhilash , Zhang Huang-Ge 
Published: January 2023

Abstract
Chemotherapy induces myeloid-derived suppressor cells (MDSCs) which promote tumor metastasis. However, the molecular basis for chemotherapy induced MDSCs remains poorly understood. Here, we demonstrate that Doxorubicin-derived MDSC promote cancer progression via their effect on angiogenesis and the formation of a premetastatic niche. miRNA-containing exosomes from doxorubicin-treated MDSC exert more potent proangiogenic effects. We also demonstrate high level of MiR-126 was detected in Doxorubicin-derived exosomes, and inhibition of miR126 encapsulated in exosomes impaired proangiogenic effects of Doxorubicin-derived MDSC. Furthermore, nanoparticle-mediated targeting delivery of anti-miR-126 to MDSCs improves Doxorubicin-based therapeutic responses, leading to inhibition of lung metastasis. Collectively, our finding provides a compelling rationale to develop therapeutic strategy for overcoming chemotherapy resistance.