bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel targeting methods for pancreatic cancer. (TECH2P.914)
Authors: Knowlton Andrea , Restuccia Antonietta , Delitto Daniel , Trevino Jose , Hudalla Greg , Wallet Shannon 
Published: January 2023

Abstract
Pancreatic cancer is the fourth leading cause of cancer deaths in the United States and one of the most difficult-to-treat cancers. Despite advances in chemotherapy, 5-year survival following surgical resection is only 23% in specialized pancreatic surgery centers. A major stumbling block for these and other cancer treatments is a way to efficiently deliver therapeutics to the target cells. Thus, we have identified high affinity molecular targeting molecules unique to an individual pancreatic cancer, which were generated utilizing a high throughput system using fresh surgical specimens along with phage display to develop novel small chain variable fragments (scFv). Here we present the characterization of two scFv molecules identified, through which we demonstrate by ELISA and BioLayer Interferometry that our scFv bind with high specificity to the neoplasm from which they were derived, while remaining minimally reactive to healthy pancreatic tissue. In addition, we have engineered these scFV to be assembled into a homotrimeric alpha-helical coiled coil using a novel platform in order to retain its multimeric nature. Future studies will assess the ability of these scFv to home to the original neoplasm using our established patient-derived tumor xenograft models whereby the human tumor is grown in NOD/SCID mice. Together these data will determine whether screening individual neoplasms using phage display can be used to develop scFv as personalized targeting molecules.
Rapid quantification of maleimide in bioconjuated samples using a novel colorimetric method (TECH2P.761)
Authors: Liao Jinfang , Guo Haitao , Luo Zhen , Zhao Qin , Diwu Jack 
Published: January 2023

Abstract
A variety of crosslinking reagents with a maleimide group are widely used for crosslinking proteins to proteins, or proteins to other macromolecules. A key challenge in maleimide crosslinking technology is the lack of rapid and sensitive methods for quantifying the amount of maleimide groups in a biopolymer. Maleimides can be directly measured spectrophotometrically at 302 nm. However, the small extinction coefficient of maleimide renders this approach insensitive. In addition, the measurement is non-specific due to the fact that proteins have absorbance at the same wavelength. To address this unmet need, a rapid colorimetric method was developed to quantify maleimide group by using a novel maleimide sensor with the maximum absorbance at ~780 nm. The maleimide sensor can react with maleimide groups with high selectivity and sensitivity. After reaction with the maleimide sensor, the maleimide-linked sample was purified through a single spin column to remove the excess sensor, and the absorption spectrum was further measured. The ratio of maleimide to protein in the maleimide-linked samples was calculated by the ratio of absorbance at 780 nm to the absorbance at 280 nm. Based on this newly developed method, number of maleimide groups can be measured rapidly and precisely for a biopolymer (such as antibodies).
Metal-conjugated neutravidin for MHC multimer assays using mass cytometry (TECH3P.939)
Authors: Ornatsky Olga , Wong Gladys , Majonis Daniel , Delcommenne Marc , Narumiya Kohei 
Published: January 2023

Abstract
Antigen-specific T cells in blood can be detected through the use of soluble MHC-peptide ligands that engage αβTCR. The fluorescent tetramer assay, developed in part by John Altman et al. (1) of the Vaccine Research Center, has become a standard tool for immunologists. Evan Newell et al. ( 2) adapted this peptide-MHC tetramer technology to mass cytometry for the purpose of screening of up to 109 different peptide-MHC tetramers in a single human blood sample, as well as analyzing another 23 markers of T-cell phenotype and function using a recombinant form of streptavidin conjugated to metal tags (Maxpar® kits, Fluidigm CA). We will describe the workflow for enumeration and identification of CMV-specific CD8+ T-cells with Neutravidin-metal reagent complexed with HLA-A*0201 CMV pp65 biotinylated monomer (MBL International, MA) simultaneously with metal-labeled surface markers, cisplatin dead-cell identifier, and the use of metal barcoding of several samples into one. There are 198 different biotinylated monomers, commercially available from MBL International, which can be combined with up to 35 isotope-tagged Neutravidin reagents to design a highly multiparametric assay. 1. Altman, J. D. et al. Phenotypic analysis of antigen-specific T lymphocytes. Science 274: 94-96 (1996) 2. Newell, E. W. et al. Combinatorial tetramer staining and mass cytometry analysis facilitate t-cell epitope mapping and characterization. Nat. Biotechnol. 31, 623-629 (2013).
Manipulating DNA damage response pathways for the treatment of experimental autoimmune encephalitis (THER7P.952)
Authors: McNally Jonathan , Millen Scott , Terrell Catherine , Carroll Katelyn , Hildeman David , Katz Jonathan , Jordan Michael 
Published: January 2023

Abstract
The race between pathogens and the immune response drives rapid expansion of lymphocyte populations, suggesting genomic stress in responding lymphocytes. We have previously reported the unique apoptotic sensitivity of activated T cells to DNA damaging drugs in diverse therapeutic contexts. We now report that activated T cells display a pronounced DNA damage response (DDR) in vitro and in vivo. We hypothesized that this intrinsic DDR could be manipulated with targeted small molecules for the elimination of pathologic T cells in a therapeutic context. We found that potentiation of p53 (via inhibition of MDM2)) or abrogation of the G2/M cell-cycle checkpoint (via inhibition of WEE1) synergized with DNA damaging drugs for the elimination of acutely activated, pathologic T cells. Moreover, a combination of p53 potentiation and checkpoint abrogation displayed therapeutic benefits in Experimental Autoimmune Encephalitis a preclinical disease model of Multiple Sclerosis, without the concurrent use of genotoxic chemotherapeutics. Additionally, combination inhibitor therapy showed minimal off target toxicities on other immune populations. Thus, targeted manipulation of p53 and cell cycle checkpoints may represent a new therapeutic modality with significant translational potential for diverse immune mediated diseases.
Engineered MBP antigen-specific human regulatory T cells: direct and bystander immunosuppressive function (THER7P.954)
Authors: Kim Yong , Zhang Ai-Hong , Wucherpfennig Kai , Scott David 
Published: January 2023

Abstract
Clinical application of expanded T regulatory cells (Tregs) offers great promise for the treatment of undesirable immune responses. However, treatment with polyclonal Tregs, which are not specific, could potentially cause global immunosuppression. To avoid this problem, we propose that antigen-specific Tregs, based on chimeric antigen receptor (CAR) therapy in leukemia, would be desirable to control autoimmune responses. We engineered a myelin basic protein-specific TCR in a retroviral vector and transduced it to human naïve CD4+ T cells and Tregs. The TCR V genes came from an MBP-specific T-cell clone derived from an MS patient (Ob2F3, Ota et al., Nature 346, 183-7, 1990). Transduced Tregs proliferated in vitro in response to MBP peptide on DR2 APC. Foxp3 and Helios expression were preserved in transduced Tregs during long-term expansion and these specific Tregs did not produce any inflammatory cytokines. Treg-unique activation markers, Foxp3 and LAP, were upregulated by MBP. Importantly, MBP-specific Tregs suppressed proliferation and cytokine production by MBP-specific T effector cells in the presence of MBP peptide. We also observed bystander suppression of MBP responses in vitro with Tregs specific for a different epitope when both peptides were present. We propose that MBP-specific Tregs can control pathologic inflammation as a future therapy in MS patients.
Unique gut commensal bacteria in CD44 deficient mice can ameliorate experimental autoimmune encephalomyelitis (THER7P.943)
Authors: Guan Hongbing , Singh Udai , Singh Narendra , Hao Haiping , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Gut commensal microbiota synergize diet, metabolites, and innate immunity to maintain homeostasis of biological system. There are now emerging evidences implicating the role of gut microbiota in elicitation of human autoimmune diseases. It is has been reported that gut commensal microbiota and myelin autoantigen cooperate to trigger autoimmune demyelination and experimental autoimmune encephalomyelitis (EAE). In the current study, we aimed to identify specific microbiota associated with the pathogenesis of EAE. We used EAE-susceptible CD44+/+ (WT) mice and EAE-resistant CD44-/- (KO) mice for the analysis of gut commensal bacteria. We found distinct bacteria profiles in naïve WT and naïve KO mice. After immunization with MOG35-55 peptide, the profiles dramatically changed on the bacterial classification levels. We identified several species as putative probiotics against the disease. We also observed these probiotics can prevent the development of EAE, and significantly ameliorate the disease. Upon analysis of the metabolites in situ in the gut, we identified probiotic-composites that suppressed encephalitogenesis of CD4+ effector T cells. Our results may offer novel strategies for the prevention and treatment of multiple sclerosis.
Plant-based product Apigenin attenuates EAE severity through the modulation of immune cell function (THER7P.948)
Authors: Ginwala Rashida , McTish Emily , Sagar Divya , Raman Chander , Jain Pooja , Khan Zafar 
Published: January 2023

Abstract
The use of Apigenin, a naturally occurring plant flavone, for centuries to treat asthma, Parkinson's disease, neuralgia, and shingles, indicates its importance in the regulation of inflammation. However, its effect on dendritic cells (DC) that maintain the critical balance between an immunogenic and tolerogenic immune response especially in neuroinflammation is relatively unknown. To test if Apigenin treatment ameliorates disease after onset of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, C57BL/6 mice immunized with MOG35-55 were treated with Apigenin. A significant reduction in severity of EAE progression was observed in the treated mice with disease peak lasting only a day. Splenocytes isolated from Apigenin treated EAE mice show decreased expression of α4 integrin and increased DCs and macrophages compared to untreated EAE mice. This correlated with immunohistochemistry findings of decreased immune cell infiltration and reduced demyelination in the CNS. In vitro, Apigenin inhibited TNF-α and IL-6 secretion in mice splenic DCs stimulated with LPS and the cell surface expression of MHC II and CD86 molecules on bone marrow derived DCs. These results indicate a protective role of Apigenin against the neurodegenerative effects resulting from the entry of DC stimulated pathogenic T cells into the CNS. Apigenin can thus serve as a potential therapy for neuroinflammatory disease through its regulation of immunogenic T cell response.
Cross reactivity of allo and xenoimmune response in transplantation (TRAN2P.964)
Authors: Byun Nari , Kim Hyun-Je , Yoon Il-Hee , Kwon Oh-Sang 
Published: January 2023

Abstract
Many type I diabetes patients are suffering from hypoglycaemia unawareness. And porcine islet transplantation is one of the promising modalities to treat the problematic condition in a physiologic manner. But by transplanting porcine islet, xenoantigen sensitization is inevitable. For clinical application of porcine islet transplantation, the knowledge of the outcome and consequent effects of xenoantigen sensitization needs to be fully established. Our focus was whether xenoantigen sensitization would influence subsequent allotransplantation in any way. We confirmed that xenoantigen sensitization does not induce alloreactive antibody. And by ELISpot assay we demonstrated that xenoreactive T cells are not responsive to alloantigen. Lastly, we confirmed that xenoantigen sensitized mice have the same graft survival day compared with non sensitized mice in allogeneic islet transplantation setting. With these results put together, we conclude that porcine antigen sensitization can not induce alloantigen sensitization in mice.
Induction of antigen specific transplantation tolerance using chimeric antigen receptor type T cells engineered to kill allospecific T cells by a gene therapy immunotherapeutic approach (TRAN2P.968)
Authors: Rajawat Yogendra , Berkson Julia , McFarland Hugh , Morgan Richard , Zhao Yangbing , Rosenberg Amy 
Published: January 2023

Abstract
Development of specific immunotherapies to prevent graft rejection and graft-versus-host disease offers the promise of selectively deleting only allospecific immune responses. Veto activity is the ability of a cell to specifically suppress/delete only T cells directed against antigens of the veto cells themselves, but not against third-party antigens. In the present study, we reasoned that providing TCR-like signaling to allogeneic major histocompatibility complex (MHC) molecules on the surface of CD8+CTLs might result in enhanced veto function. To generate the donor antigen specific veto cells, we have utilized the Chimeric Antigen Receptor (CAR) T cell approach, in which CD8+CTLs express an MHC alloantigen fused to signaling molecule apparatuses. We fused mouse MHC class I alloantigen (H-2Dd) to the hinge and trans-membrane domains of the mouse CD8 chain plus intracellular signaling sequences derived from mouse 4-1BB, and CD3-ζ chain creating various CAR constructs. These CARs were cloned into retroviral vectors. Ex-vivo transduced T cells (FVB-Dd) were adoptively transferred in the recipient parental mice strain (FVB-H-2q) which do not express Dd. Two weeks post-adoptive transfer, we detected low levels of Dd+CD8+ T cells in lymph node and spleen, ranging from 0.2-1.2% of total CD8+ T cells. Veto activity of these Dd+CD8+T cells will be examined by in vitro Dd specific CTL assays, proliferation assays, and rejection of Dd skin grafts in parental strain (FVB) mice.
Accelerated progression of Hodgkin’s-like lymphoma in golli-deficient mice is associated with higher induction of CD4+FoxP3+ regulatory T cells (TUM9P.1018)
Authors: Feng Ji-Ming , Ouyang Suidong , Han Guangming 
Published: January 2023

Abstract
The spontaneously occurred lymphomas in SJL/J mice have many pathological features similar to Hodgkin’s lymphoma in human. The malignant growth of the tumor cells is dependent on the support of host FoxP3+ CD4+ regulatory T cells (Tregs). In this study, we report that the golli protein, a negative regulator of CRAC (calcium release activated calcium) channel in T cells, plays a critical role in the tumorigenesis of Hodgkin’s-like lymphoma. The ablation of golli protein in SJL/J mice results in an accelerated progression of Hodgkin’s-like lymphoma which is associated with a facilitated induction of FoxP3+ Treg cells. Our results suggest that through regulating the induction of Treg cells, golli protein affect the progression of Hodgkin’s-like lymphoma.
NKG2D-NKG2D ligand interaction inhibits the outgrowth of low-grade B cell lymphoma (TUM9P.1019)
Authors: Markiewicz Mary , Raju Saravanan , Kretzmer Lena , Koues Olivia , Payton Jacqueline , Oltz Eugene , Cashen Amanda , Polic Bojan , Schreiber Robert , Shaw Andrey 
Published: January 2023

Abstract
Transgenic mice we generated with ubiquitous expression of the NKG2D ligand RAE1ε, and therefore low surface NKG2D expression, developed natural, spontaneous B cell lymphoma at a younger age than wild-type mice. Transplantation of lymphomas from RAE1ε transgenic mice into lymphodeficient mice resulted in expression of NKG2D on immune cells transferred along with the tumor and rejection of the lymphoma cells, suggesting NKG2D is important in controlling lymphoma outgrowth in wild-type mice. In the absence of overt tumor masses, older mice developed lymphoid aggregates in non-lymphoid organs and these aggregates occurred at a younger age in RAE1ε transgenic and NKG2D-deficient mice. The increased incidence of both these lymphoid aggregates and tumor masses in RAE1ε transgenic mice suggests these lymphoid aggregates are slow growing indolent lymphomas. Although NKG2D ligands were undetectable on wild-type tumors, ligands were present on splenic B cells in a similar proportion of 10 month-old wild-type and NKG2D-deficient mice as those with lymphoid aggregates present. These results suggest that 1) induction of NKG2D ligands is an early marker of B cell transformation, 2) NKG2D-NKG2D ligand interaction suppresses B cell lymphoma growth, and 3) loss of NKG2D receptor-ligand interactions leads to the transformation of indolent B cell lymphomas into aggressive tumors in this model. We are now exploring whether these results are relevant to human follicular lymphoma progression.
Inhibition of Treg suppression by GITR stimulation causes tumor regression selectively in the context of a pro-inflammatory tumor microenvironment (TUM9P.1005)
Authors: Zappasodi Roberta , Budhu Sadna , Decicco Emily , Schild Tanya , Draleau Kyle , Schaer David , Yang Xia , Merghoub Taha , Wolchok Jedd 
Published: January 2023

Abstract
GITR stimulation as immunotherapy has become increasingly important given the recent success of immune modulation in cancer therapy. As anti-GITR therapy is translated into clinic, defining the basis of its activity and failure is key to identify prognostic markers and more effective combination therapies. We thus studied how GITR stimulation affects T cells in B16 melanoma-bearing mice treated in curative (early) or refractory (advanced) condition with the agonist antibody DTA-1, and verified their functional changes in vitro. In vivo, Treg frequency was increased, Teff activation reduced and tumor antigen presentation impaired in more advanced tumors. DTA-1 reduced Treg frequency and expression of suppression markers in curative and refractory conditions. In contrast, Teff activation and tumor-associated MHCs were induced only in curative conditions. In vitro, DTA-1-pretreated Treg lost their suppressive activity without downregulating either Foxp3 or Helios, and increased Th1/Th17 cytokine release. Interestingly, co-culture with T cells was sufficient to up-regulate MHC-II on B16, but MHC-I was induced if both T cells and DTA-1 were present. GITR stimulation on human Tregs via GITRL consistently abrogated their functions, thus validating GITR as a target to revert immune tolerance also in this setting. Our data suggest that Treg inhibition by anti-GITR therapy should be exploited in combination with treatments promoting Teff functions and/or tumor killing in future trials.
Ptnp11 deletion in CD4+ cells causes cartilage tumors in a T cell independent manner (TUM9P.1006)
Authors: Miah S. M. Shahjahan , Salter Alexander , Reilly Emma , Fugere Celine , Jayasuriya Chathuraka , Yang Wentian , Chen Qian , Brossay Laurent 
Published: January 2023

Abstract
The ubiquitously expressed tyrosine phosphatase SHP-2 (encoded by Ptpn11) is required for constitutive cellular processes including proliferation, differentiation, and the regulation of immune responses. During development and maturation, subsets of T cells and iNKT cells express a variety of inhibitory receptors known to associate with phosphatases, which in turn, are thought to dephosphorylate key players of TCR mediated activation. Using mice conditionally deficient for SHP-2in the T cell lineage, we show that classical T cells and iNKT cells develop normally in the thymus and exhibit proper maturation and functions in the periphery. These results suggest that SHP-2 serves a redundant role with other tyrosine phosphatases in the T cell lineage and is dispensable for T cell function. Unexpectedly, in aging mice, Ptpn11 gene deletion driven by CD4 Crerecombinase leads to cartilage tumors in a T cell independent manner, indicating that SHP-2 is a tumor suppressor.
Transcription factor slug-upregulated microRNA-221 suppresses E-cadherin expression and promotes breast cancer progression (TUM10P.1031)
Authors: Liu Yuan , Pan Yi , Gu Hongwei , li jing 
Published: January 2023

Abstract
E-cadherin protein expression is blocked during tumorigenesis due to slug/snail-mediated suppression of E-cadherin transcription. However, this mechanism of transcriptional blockade cannot explain why metastatic breast cancer cells maintain very low level of E-cadherin even with E-cadherin mRNA overexpression. In the present study, we reveal that E-cadherin is also post-transcriptionally regulated by slug-promoted microRNA-221 (miR-221), which serves as an additional lock preventing E-cadherin expression in metastatic cancer cells. Profiling the predicted E-cadherin-targeting miRNAs in breast cancer tissues and cell lines treated with/without TGFβ shows that the miR-221 level is significantly higher in metastatic breast cancer tissues and in MDA-MB-231 cells compared with the cancer-adjacent normal tissues or non-metastatic MCF-7 cells, and that miR-221 level is subjected to the TGFβ regulation. The level of miR-221 is inversely correlated with E-cadherin expression in breast cancer cells, and experimentally depleting or elevating miR-221 induces or decreases E-cadherin expression, consequently promoting or suppressing breast cancer progression. Moreover, we found that miR-221 suppresses E-cadherin translation via binding to the open reading frame of the E-cadherin mRNA and that miR-221 is specifically upregulated by Slug. In conclusion, our results demonstrate for the first time that Slug-upregulated miR-221 promotes breast cancer via targeting E-cadherin.
The NK killer receptor Ncr1 shapes tumor architecture to prevent metastasis (TUM10P.1034)
Authors: Glasner Ariella , Mandelboim Ofer 
Published: January 2023

Abstract
Natural killer (NK) cells were so-named because of their excellent ability to kill tumors and pathogens. NKp46 (Ncr1 in mice) is a prominent NK killer receptor, critically involved in controlling various pathogens as well as tumors and tumor metastases. We and others and have demonstrated the ability of the NK killer receptor Ncr1 to contain tumor growth, however, the mechanisms underlying the ability of Ncr1 to control tumor growth and metastases in vivo are yet undetermined Metastatic melanoma is an extremely aggressive disease and the leading cause of death from skin cancers. Here, we used two melanoma spontaneous metastases models: B16F10.9 (B16) and RET, the Ncr1 knockout mice (Ncr1gfp/gfp), Ifnγ-/- and Tnfα-/- knockout mice to visualize the tumors’ architecture in-vivo, using Reflectance Confocal Microscopy (RCM). We demonstrate that Ncr1, through Ifnγ prevents tumor metastasis by modulating the structural properties of the developing tumors, at the level of the tissue organization. Using RNAseq we have analyzed the transcriptome of Ifnγ treated B16 cells and reveled a 24 fold increase in the integrin binding ECM protein Fibronictin1 (FN1), the absence of which was reported to correlate with increased metastases. The finding that Ncr1, through Ifnγ, shapes the tumors’ structures and thus controls metastasis may lead to generation of new drugs aimed at targeting specifically the tumors’ architecture and new treatment options for melanoma patients.