bluuebunny/crossref_metadata_2025_split huggingface dataset:
Combination B-lymphocyte Depletion Treatment with Anti-CD20 and BAFF Blockade is No Superior to BAFF Blockade Alone in The NOD Model of Type 1 Diabetes
Authors: Wang Qiming , Serreze David V 
Published: January 2023

Abstract
While type 1 diabetes (T1D) is an autoimmune disease in which autoimmune destruction of pancreatic β cells is ultimately mediated by T-cells, in the NOD mouse model, and also likely humans B-lymphocytes also play a key pathogenic role. B-lymphocyte depletion either through antibody targeting CD20 or blocking their access to BAFF elicits strong T1D protection in NOD mice. However, whether the combined strategy of targeting both CD20 and BAFF could further improve therapeutic efficacy for T1D remained unknown. Currently humans at high future T1D risk can only be identified at late prodromal stages of disease indicated by markers such as insulin autoantibodies (IAA). We show BAFF blockade treatment commenced in already IAA positive NOD mice prevented progression to overt T1D, whereas anti-CD20 was ineffective as a mono- or co-therapy at this disease stage. While BAFF blockade depletes the majority of islet infiltrating B-lymphoctes, a memory-like subset remained, whose potential diabetogenic capacity needs further exploring. Although BAFF blockade treated mice are protected from T1D development, the severity of insulitis was not reduced. One complicating factor underlying the failure of anti-CD20 is that NOD mice developed neutralizing antibodies against the injected anti-CD20, and the degree of B-lymphocyte rebounding correlates with the titer of neutralizing antibodies present in the sera. The dual directed immunotherapy does not either improve the protection offered by BAFF blockade only, or reduce the severity of insulitis, despite the greater depletion on various B-lymphocyte subsets and further reduction on those that survived of MHC and CD40 molecules respectively involved in antigen presentation, and class switching.
Identification and characterization of an antagonistic anti-mouse CD40 antibody
Authors: Ralph Kerry L. M. , Panzenbeck Mark , Xiao Haiguang , Frego Lee , Bigwarfe Tammy , Waltz Erica , Grimaldi Christine , Last-Barney Kathy , Souza Don , Brodeur Scott , Nabozny Gerald , Fine Jay S. , Distler Jorg H. W. , Ramanujam Meera 
Published: January 2023

Abstract
Targeting CD40-CD40L interactions has been an interesting drug concept for the treatment of autoimmune diseases given this pathway’s role in the development of both humoral and cell mediated immune responses. While preclinical blockade of CD40L is well documented in various autoimmune animal models of disease, limited data exists for CD40 blockade due to lack of a truly antagonistic anti-mouse CD40 tool antibody (Ab). Here we describe the in vitro and in vivo characterization of a fully antagonistic anti-mouse CD40 monoclonal Ab (BI CD40-1); a chimeric rat Fv anti-mouse CD40 mAb engineered with a mouse IgG2a Fc containing mutations to abrogate Fcγ receptor binding. BI CD40-1 blocks molecular CD40-CD40L interactions (IC50 = 0.25 nM) and exhibits potent binding to CD40 expressed on mouse B cells (EC50 = 0.42 nM ± 0.08). In vitro profiling of BI CD40-1 using a mouse splenocyte proliferation assay confirmed potent antagonistic activity (IC50 = 0.27 nM ± 0.09) as well as absence of any agonistic properties (stimulation index &lt; 2 @ 67 nM). Administration of BI CD40-1 to mice prior to ovalbumin (OVA) immunization resulted in dose-dependent blockade of OVA-specific IgG responses (100, 100, 74, 0% inhibition at 10, 3, 1, and 0.3 MPK; day 13) correlating with similar dose dependent receptor occupancy and inhibition of B cell activation as measured by ex vivo CD54 upregulation. Prophylactic dosing in cGVHD model of scleroderma showed dose dependent protection in disease development as seen by decreased dermal thickness, myofibroblast counts and collagen deposition. BI CD40-1 represents a novel fully antagonistic anti-mouse CD40 mAb, an important tool for mechanistic understanding of the therapeutic value of targeting of CD40 in inflammatory diseases.
Molecular interactions of small molecule inhibitors targeting cytoplasmic TIR domains.
Authors: Snyder Greg A , Irwin Gregory , Mistry Pragnesh Dilipkumar , VanBesien Richard , Flecther Steven , MacKerell Alexander , Brown Lindsey , Xiao Tsan Sam , Wintrode Patrick , Vogel Stefanie 
Published: January 2023

Abstract
Regulation of Toll-like receptor (TLR) signaling using small molecule agonists and antagonists have been widely sought after for controlling inflmation and disease. Most studies have focused on therapeutic targeting of TLR extracellular ligand binding domains. Recently, Computer-aided drug design (CADD) screens specifically targeting intracellular Toll-Interleukin-1 receptor resistance domains have identified and functionally characterized several small molecule and peptide inhibitors which protect against lethality in animal models of infection, inflammation and disease. We have sought to structurally characterize the molecular interactions of CADD derived small molecule inhibitors with respective TLR-TIR domains by determining the X-ray co-crystal structures of TLR2 –TIR domain in the presence of C29 and its substructure O-vanillin. Positive electron density is observed near the BB loop and residue Ile 685 of native TLR2-TIR in the presence of the small molecule inhibitor (C29) exhibit compared with the native apo form of this structure. However, the presence of (S-(DIMETHYLARSENIC) CYSTEINE) in this crystallization condition complicate analysis. To rule out the effects of DMSO and (S-(DIMETHYLARSENIC) CYSTEINE) we sought to examine small molecule inhibitor C29L substructure (o-Vanillin) in a second crystal form of TLR2 and which contains a more simplified crystallization condition grown in the presence of 1mM of C29L. This co-crystal exhibits positive electron density located in an around the BB loop compared with apo forms of this structure. Additionally, in this recent structure form we observe additional DD loop density previously not defined in the originally reported crystal form.
IFN-β suppresses GM-CSF production by CD4+ T cells by inhibiting STAT5 signaling
Authors: Rasouli Javad , Mari Elisabeth Rose , Ciric Bogoljub , Rostami Abdolmohamad 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an autoimmune disease characterized by accumulation of immune cells in the central nervous system (CNS). Granulocyte-macrophage colony-stimulating factor (GM-CSF) is necessary for development of CNS autoimmunity in animal models of MS, suggesting that GM-CSF also plays an important role in the pathogenesis of MS. We previously reported that untreated MS patients have higher frequencies of GM-CSF+ CD4+ T cells in their peripheral blood, and that IFN-β therapy normalized their numbers. Since IFN-β suppresses GM-CSF expression by CD4+ T cells in MS, we sought to determine the mechanisms of its action. It has been shown that IFN-β activates multiple JAK/STAT signaling pathways in T cells, and that GM-CSF expression in T cells is largely dependent on STAT5 signaling. Here, we show that in vitro IFN-β strongly suppressed GM-CSF production by T cells from healthy donors, untreated and IFN-β-treated MS patients, as well as patients diagnosed with other neurological diseases. IFN-β did not increase T cell apoptosis in our cell culture; hence, reduced production of GM-CSF by T cells was not due to pro-apoptotic effects of IFN-β. IFN-β significantly decreased the phosphorylation of STAT5 in total CD4+ T cells, including GM-CSF-producing CD4+ T cells. In summary, our findings suggest that IFN-β suppresses GM-CSF production by CD4+ T cells by inhibiting STAT5 signaling.
FGL2 promotes tumor progression via inducing TIGIT expression on T cells in tumor microenvironment of glioma
Authors: Yan Jun , Gabrusiewicz Konrad , Xia Xueqing , Heimberger Amy B. , Li Shulin 
Published: January 2023

Abstract
Glioblastoma multiforme (GBM) is the most common primary brain tumor with a median survival of only 14.6 months. Our previous work shows that FGL2 promotes glioma tumor growth and the expression levels of FGL2 positively correlated with glioma grade in patients. T cell Ig and ITIM domain (TIGIT) is an inhibitory receptor expressed by activated T cells, Tregs, and NK cells. Here we found that there is a statistically significant positive correlation between FGL2 and TIGIT mRNA expression level in both low grade glioma database (TCGA, provisional) and glioblastoma multiforme database (TCGA, provisional). Future, FGL2 and TIGIT are coordinately expressed by human tumor-infiltrating lymphocytes of glioma as detected by flow cytometry. In addition, TIGIT expression on T cells which suppresses their function, can be induced by initial T cell activation or FGL2 in vitro. Induced TIGIT expression upon initial T cell activation is suppressed in FGL2KO T cells which has higher tumor-killing ability than WT T cells. Therefore, our results show that FGL2 induces TIGIT expression in brain-infiltrated lymphocytes which suppresses anti-tumor immune response in glioma. By this mechanism, FGL2 promotes tumor progression of glioma.
Role of Prostate apoptosis response-4 tumor suppressor in the survival and growth of Chronic Lymphocytic Leukemia
Authors: McKenna Mary Kathryn , Nooti Sunil K , Alhakeem Sara Samir , Gachuki Beth W , Frissora Frank , Greene Joseph T , Byrd John C , Muthusamy Natarajan , Rangnekar Vivek R , Bondada Subbarao 
Published: January 2023

Abstract
Chronic Lymphocytic Leukemia (CLL), the most common adult leukemia in the western world, is characterized by accumulation of clonally expanded CD5+CD19+ B lymphocytes in blood and secondary lymphoid organs with impaired apoptotic mechanisms. Prostate apoptosis response-4 (Par-4) is a pro-apoptotic tumor suppressor protein, which is silenced by promoter methylation in more than 30% of all cancers. It is also secreted and induces apoptosis selectively in many types of cancer cells but not in normal cells. Here we characterized the role of Par-4 in CLL cells using a murine model. The Eμ-Tcl1 mouse serves as an excellent model to study CLL as they develop a CLL like disease by 9–13 months of age, due to B cell specific over-expression of the oncogene, T cell Leukemia 1 (Tcl1). Adoptive transfer of primary CD5+CD19+ CLL cells from the Eμ-Tcl1 CLL mice into recipient syngeneic mice leads to the development of a CLL like disease within 3–5 weeks of transfer. Surprisingly Eμ-Tcl1 CLL cells constitutively express more Par-4 than normal B-1 or B-2 cells in mice. These CLL cells also secrete Par-4 which is functional in being cytotoxic to prostate cancer cells. We showed that Eμ-Tcl1 CLL cells have constitutively active B-cell receptor signaling and that inhibition of BCR signaling causes a decrease in Par-4 expression and increases apoptosis. Interestingly, we have found that shRNA mediated knockdown of Par-4 in human CLL cell lines, results in their reduced growth. These results suggest that intrinsic Par-4 may play a pro-survival rather than pro-apoptotic role in CLL. We are currently investigating the mechanisms underlying this novel role of Par-4 and regulation of its expression in CLL cells using CRISPR system to knockout Lyn and Btk in CLL cells.
PD-1 expression is not indicative of T cell exhaustion in breast cancer tumors.
Authors: Egelston Colt , Avalos Christian , Lee Peter P. 
Published: January 2023

Abstract
PD-1 expression on tumor infiltrating lymphocytes (TILs) has been implicated to be associated with T cell exhaustion. Indeed, we have found that PD-1 is expressed on a majority of both CD4 and CD8 T cells in the tumors of breast cancer patients. To determine if patient T cells have an exhausted functional phenotype, we compared patient peripheral blood mononuclear cells (PBMCs), tumor draining lymph nodes (TDLNs), TILs, and healthy donor PBMCs. We examined the cytokine production capacity in response to non-specific stimulation, followed by intracellular flow cytometry for production of IFNγ and TNFα. In the majority of patients, we found TIL memory T cells at least as potent as circulating memory T cells in producing both IFNγ and TNFα. Furthermore, PD-1+ T cells did not show any significant reduction in cytokine production compared to PD-1− T cells. Additionally, immune phenotyping by flow cytometry shows a lack of other checkpoint molecules, such as LAG-3, TIM-3, and CTLA-4 on TILs. This data suggests that PD-1+ T cells in breast cancer tumors may not suffer from exhaustion and that PD-1 expression alone does not indicate T cell exhaustion.
The adhesion molecule PSGL-1 is a checkpoint regulator that promotes T cell dysfunction in melanoma tumors
Authors: Tinoco Roberto , Carrette Florent , Barraza Monique L , Bradley Linda M 
Published: January 2023

Abstract
The tumor microenvironment can promote tumor growth by suppressing effector T cell responses. Using a melanoma model that recapitulates aspects of human disease, we investigated the role of the adhesion molecule, P-selectin glycoprotein ligand-1 (PSGL-1) in tumor infiltrating lymphocytes (TILs). We found that PSGL-1 was highly expressed on TILs that also expressed high PD-1 and TIM-3 inhibitory receptors. PSGL-1-deficiency did not impact T cell recruitment in tumors but instead resulted in a greater TIL accumulation. Furthermore, PSGL-1-deficient T cells had reduced PD-1 expression and increased production of effector cytokines that resulted in delayed tumor growth. We did not find differences in Treg recruitment but instead had an increased effector to Treg ratio in tumors. Our findings show that PSGL-1 does not have a nonredundant role in the trafficking of TILs into melanoma tumors, but can act as an immune checkpoint inhibitor to limit anti-tumor T cell responses in the tumor microenvironment thereby promoting tumor growth.
Inflammasome-independent role of AIM2 in mediating protection against colorectal tumorogenesis
Authors: Karki Rajendra , Man Si Ming , Zhu Qifan , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
Colorectal cancer is a leading cause of cancer-related deaths. Mutations in the innate immune receptor AIM2 are frequently identified in patients with colorectal cancer. AIM2 inflammasome is activated by double stranded DNA and inflammasomes are central regulators of intestinal homeostasis. The expression of the AIM2 is reduced in several types of tumors compared with healthy tissues, highlighting a previously undefined function for this sensor in the development of colorectal cancer. Here, we found that Aim2-deficient mice were hypersusceptible to colonic tumor development. Production of inflammasome-associated cytokines and other inflammatory mediators were largely intact in Aim2-deficient mice, however, intestinal stem cells were prone to uncontrolled proliferation. Aberrant Wnt signaling expanded a population of tumor-initiating stem cells in the absence of AIM2. Therapeutic modulation of AIM2 expression has the potential to prevent colorectal cancer.
The role of Signal Transducer and Activator of Transcription protein 5 (STAT5) in non-small cell lung cancer (NSCLC)
Authors: Friedrich Juliane Michaela Margarete M , Heim Lisanne , Springel Rebekka , Mittler Susanne , Gafencu Dumitrita , Hartmann Arndt , Sirbu Horia , Finotto Susetta 
Published: January 2023

Abstract
Signal Transducer and Activator of Transcription protein 5 (STAT5a/b) belongs to a family of T cell transcription factors. The different isoforms of STAT5 are activated by several cytokines and tyrosine kinases, which led to the assumption, that both STAT5 isoforms regulate cell growth. Interleukin (IL)-2, a major cytokine regulating immunocompetent cell survival, proliferation and differentiation can activate STAT5. Interestingly, by contrast to other T cell subsets, T regulatory cells (Tregs) express the IL-2 receptor constitutively. It is considerable that Tregs are immunosuppressive and thus are able to suppress the function of certain effector T cells, eg CD4, CD8 and natural killer T cells. Tregs are highly dependent on IL-2 and thus are able to consume IL-2 more efficiently than effector cells, which results in a depletion of IL-2 from effector cells and leads to a disorder in the immune response. We thus asked about the role of STAT5 in NSCLC.
We measured increased STAT5 protein levels by Western Blot in lung tumor tissue compared to control lung tissue derived after lung surgery from patients who suffered from lung cancer, especially adenocarcinoma (ADC). This finding was confirmed by STAT5 immunohistochemistry (IHC) staining, where a stronger signal for STAT5 was detected in lung tumor tissue compared to lung control region, in a cohort of ADC patients (n=10). Moreover, preliminary FACS analysis of freshly isolated cells from the resected lung of one patient with NSCLC showed strongly upregulated pSTAT5 levels in tumor infiltrating CD4+ T lymphocytes compared to CD4+ lymphocytes derived from respective lung control region.
Thus unrevealing the role of STAT5 in lung tumor should help to improve lung cancer immunotherapy.
Inflammasome/IL-1 pathways in tumor microenvironments
Authors: Guo Beichu , Zhang Jinyu , Fu Shunjun 
Published: January 2023

Abstract
Emerging evidence indicates that chronic Inflammation plays important roles at all stages of tumor development including initiation, growth, invasion and metastasis. IL-1β and other IL-1 family members are major mediators in an inflammatory response. Clinical studies reveal increased concentration of IL-1β protein in tumor tissues, and such increase is associated with poor prognosis for cancer patients. While chronic inflammation is associated with tumor development, the mechanism of such connection remains not completely understood. Our preliminary data have demonstrated that inflammasome and IL-1β play a critical role in promoting tumor growth and metastasis. An inflammasome is a multimolecular complex, composed of a NOD-like protein (NLR), the adaptor protein ASC, and caspase-1, which is essential for the cleavage of pro-IL-1β into its active form. Using orthotropic breast cancer and transgenic mouse models, we found that breast cancer was associated with inflammasome activation. Our results show that mice deficient for inflammasome components had much slow tumor growth and lung metastasis. Compared to wildtype mice, inflammasome deficient mice have low levels of IL-1 in primary tumor tissues and metastastic lungs. Furthermore, our data reveal that pharmacologic disruption of inflammasome/IL-1β pathway diminished tumor growth and metastasis in vivo. These results suggest that the activation of inflammasomes and production of IL-1β in vivo may provide an inflammatory tumor microenvironment to promote tumor growth and metastasis.
MUC1 promotes an inflammatory microenvironment aggravating colitis-associated tumorigenesis in mice through up-regulation of pro-inflammatory cytokines.
Authors: Cascio Sandra , Faylo Jacque , Sciurba Joshua , Xue Jia , Finn Olivera J 
Published: January 2023

Abstract
MUC1 is a transmembrane glycoprotein aberrantly expressed in human adenocarcinoma as well as chronic inflammatory conditions. We previously demonstrated that the tumor form of MUC1, in association with p65, upregulates expression of pro-inflammatory cytokines, including IL-6 and TNF-alpha, by modulating their transcription in epithelial cancer cells. Using azoxymethane/dextran sulfate sodium (AOM/DSS) murine model of inflammation driven colon carcinogenesis, we explored mechanism underlying MUC1/p65-induced transcription of IL-6 and TNF-alpha and its significance for the microenvironment of colitis-associated cancer. MUC1.Tg mice showed higher tumor incidence, decreased survival, greater body weight loss and shorter colon length. Consistent with our previous in vitro data, expression of NF-kB-dependent cytokines was higher in MUC1.Tg mice compared to WT. Moreover, we discovered that MUC1/p65 complex modulates IL-6 and TNF gene expression by regulating the crosstalk of post-translational modifications on their promoters. To understand the significance of MUC1/p65-modulated cytokines in progressive colitis that gives rise to colon cancer, we analyzed infiltration of inflammatory cells into the inflamed colon tissues. ELISA assay and gene expression analyses demonstrated that the treatment with AOM/DSS in the presence of human MUC1 on the colonic epithelia resulted in a significant upregulation of M1 type macrophage-associated genes, including IL-6, TNF-alpha and iNOS, whereas expression of M2 type macrophage markers, such as Arginase 1, Ym1 and IL-10, were drastically reduced compared to WT mice. Thus our findings reveal a pro-inflammatory role for MUC1 in colitis-related carcinogenesis.
Brain Slice Culture Demonstrates Expedited Cell Traficking in the Presence of Fractalkine in a Medulloblastoma Tumor Microenvironment Model
Authors: Tomaszewski William H. , Dorand Rodney Dixon , Petrosiute Agne , Huang Alex 
Published: January 2023

Abstract
20% of childhood brain tumors are medulloblastomas (MB), and the five-year survival rate can be as low as 60%. The interaction between MB and native immune cells are not well understood, and identifying methods of crosstalk could produce better outcomes for patients. Fractalkine (FKN) is a ligand for the receptor CX3CR1 that plays an inhibitory role in the brain’s immune system by keeping microglia quiescent. Normally, FKN is surface bound and expressed by neurons and astrocytes, but under inflammatory conditions FKN is secreted to serve as a chemoattractant. Preliminary work in our lab shows that FKN is produced by MB and may act as a decoy to maintain microglial quiescence leading to immune escape. Therefore we investigated FKN signaling and microglial kinetics ex vivo utilizing organotypic brain slice culture coupled with real time 2-photon laser scanning microscopy. To differentiate microglia from MB, CX3CR1+/GFP or CX3CR1GFP/GFP knock-in mice were sacrificed, and the brain was removed and sectioned. Slices were collected and cultured on inserts in an incubation chamber during imaging. To determine baseline activity, CX3CR1+/GFP and CX3CR1GFP/GFP brain slices were imaged over 48 hours. MB tumor cells, labelled with SNARF-1, were added to the cultures and imaged over the same time course. Images were processed to differentiate the motility of microglia and tumors. We found that microglia from CX3CR1+/GFP moved slower than microglia from CX3CR1GFP/GFP at baseline and in the presence of tumor. Furthermore, we found that MB moved faster in CX3CR1GFP/GFP slices, suggesting that fractalkine plays a role in the migration of MB.