bluuebunny/crossref_metadata_2025_split huggingface dataset:
Improving E. coli-based anti-cancer bacterial therapeutics
Authors: Chen Noah , Danino Tal , Arpaia Nicholas 
Published: August 2023

Abstract
Bacteria-based cancer immunotherapy has existed since Dr. William Coley employed deliberate bacterial infection of tumors over a century ago. Bacteria are inherently immunogenic, self-replicating therapeutics which can be tuned to deliver various payloads, modify specificity, and decrease side effects. Though bacteria are intrinsically capable of selectively colonizing tumor tissue in mouse models, less is understood about the mechanisms behind such colonization. This is especially important as early clinical trials of anti-cancer bacterial therapeutics have been stymied by low colonization rates in human patients. This study aims to identify mechanisms by which E. coli evade clearance by the immune system specifically within tumors. Currently, we have screened E. coli mutants for tumor colonization efficiency by assessing strain prevalence after recovery from tumors in immunocompetent C57BL/6 mice. We have identified pathways which either promote or inhibit E. coli tumor colonization and validated these with competitive infection experiments. This work paves the way for greater understanding of what bacterial mechanisms allow E. coli to home to the tumor niche, which may have implications for both developing anti-cancer bacterial therapeutics as well as for understanding how bacteria colonize solid tumors.
Supported by grants from NIH (R01 CA249160, T32 GM145440)
The CXCR3 chemokine receptor is critical for the protective immune response to Plasmodium parasite infection of the liver
Authors: Reynolds Laura M , Blyn Rebecca , Wegmair Lisa , Lewis Patrick , Sheikh Amina , Okolo Vera , Wilder Brandon , Kappe Stefan , Minkah Nana 
Published: August 2023

Abstract
No highly efficacious anti-malaria vaccine exists, but whole parasite immunization (WPI) where hosts receive live attenuated Plasmodium parasites that arrest in the liver is the only strategy that confers complete protection against future Plasmodium parasite infection. Liver-resident memory CD8 T cells (TRM) that express the chemokine receptor, CXCR3, are essential for WPI-induced protection. Yet, whether CXCR3-dependent signaling plays a functional role in anti-parasite TRM immunity is not fully understood. RNA-seq analyses on hepatocytes isolated from Plasmodium-infected mice showed an upregulation of the CXCR3-dependent chemokines, CXCL9 and CXCL10. We examined protection in WT and CXCR3−/− mice upon WPI and report that in contrast to WT animals, CXCR3−/− mice exhibit impaired control of a parasite challenge given 40 days after immunization. Next, we set out to enumerate T cell recruitment and TRM numbers in the liver after WPI. CXCR3−/− mice exhibited reduced CD8 T cell numbers early after immunization yet by day 40 contained equivalent TRM numbers as compared to immunized WT counterparts. We hypothesize that while CXCR3 signaling is critical for early recruitment of activated CD8 T cells into the Plasmodium-infected liver, other chemokines compensate for the loss of CXCR3 ensuring that equivalent numbers of CD8 T cells are eventually recruited into the liver. However, as CXCR3−/− mice exhibit impaired control of infection despite containing equivalent TRM numbers, the CXCR3 axis must play other unidentified roles. Immune cells are known to be asymmetrically positioned in the liver. Ongoing studies will determine if CXCR3 is critical for the optimal positioning and function of liver TRMs after parasite infection.
Identification and characterization of CXCL13 producers in bone tissue
Authors: Okawa Takuma , Nagai Motoyoshi , Nakata Kazuaki , Dohi Taeko , Kawamura Yuki I , Hase Koji 
Published: August 2023

Abstract
The bone serves as a reservoir for several immune cell subsets in response to the low-energy status such as fasting. We previously demonstrated that naïve B cells transiently migrate from Peyer’s patches to the bone marrow during fasting in mice. This B-cell dynamics is attributed to upregulation of CXCL13 in the bone during fasting. However, CXCL13 producers in the bone remain to be determined. We observed that lineage-negative population from the bone is responsible for Cxcl13 expression. To further clarify the CXCL13 producers, we performed single-cell RNA-sequencing of bone linage-negative cells from Ad libitum-fed mice and fasting mice. UMAP analysis manifested that the numbers of fibroblasts and multipotent stem cells (MSCs) were increased in the fasting group. Interestingly, Cxcl13-producing cells were identified as osteogenic-MSCs characterized by expressions of Sca-1 (immature cells marker), Sema3c, and Dcn (osteogenic cells maker). This population also expressed an inflammatory cytokine Il6. Meanwhile, mature osteoblasts and chondrocytes expressed neither Cxcl13 nor Il6. Furthermore, co-culture with the plasma from fasting mice upregulated CXCL13 expression in osteogenic-MSCs in vitro, suggesting that a soluble factor(s) in the circulation induces CXCL13. Taken together, osteogenic-MSCs in the bone may sense nutritional status and produce CXCL13 to regulate naïve B-cell dynamics along the gut-bone axis. The pathophysiological role of CXCL13-producing osteogenic-MSCs is currently under investigation.
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The TNF superfamily cytokine receptor Death Receptor 3 is a costimulatory molecule for iNKT cells
Authors: Liman Nurcin , Park Jung-Hyun 
Published: August 2023

Abstract
Invariant NKT (iNKT) cells are a small population of thymus-generated T lymphocytes with innate-like phenotype and function that play a disproportionally important role in immune regulation and surveillance. iNKT cells are generated upon strong agonistic TCR signaling by glycolipid-loaded CD1d molecules, which differs from conventional T cells that are positively selected by weak TCR engagement with peptide-loaded MHC molecules. Whether iNKT cells are also distinct in their costimulatory requirement, however, has remained largely unresolved. Here, we report that the TNF superfamily receptor death receptor 3 (DR3) plays a previously unappreciated costimulatory role in peripheral iNKT cell activation. We found that the co-injection of agonistic anti-DR3 antibodies significantly augmented the activation and proliferation of mature iNKT cells in α-GalCer injected mice. Moreover, iNKT cells of α-GalCer and anti-DR3 co-injected mice dramatically increased their expression of proinflammatory cytokines, resulting in severe thymic atrophy. The DR3 costimulatory effect was strictly dependent on iNKT cells, because iNKT-deficient Traj18−/− mice were refractory to α-GalCer and anti-DR3 costimulation-induced inflammation. Because DR3 ligation alone did not suffice to activate iNKT cells, and DR3 exerted its effects only in the context of TCR costimulation, these results strongly support DR3 being a new bona fide costimulatory molecule for iNKT cells. Considering the interest in utilizing iNKT cells in adoptive T cell therapies and as anti-tumor bioreagents, employing DR3 as a tool to boost iNKT cell immunity will open new venues in translational and clinical research.
This work was supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.
Interferon regulatory factor 5 involves immune responses through regulation of ly6c expression
Authors: Heo Sun-Hee , Park Suk Young , Kim Na Hyun , Song Eunkyeong , Lee Sei Won 
Published: August 2023

Abstract
Chronic obstructive pulmonary disease (COPD) is characterized by impaired lung function, prolonged airflow obstruction. COPD is mainly induced by long-tern exposure to toxic agents such as particles from cigarette smoke (CS). Interferon regulatory factor 5 (IRF5) is a key transcription factor related to many inflammatory diseases. IRF5 has been involved in polarizing macrophages and recruiting neutrophiles during various conditions. The purpose of this study was to assess immune responsiveness of IRF5 in lung disease. We generated mice deficient in the Irf5 gene (Irf5−/− mice). IRF5 expression was detected in lung and spleen of wild type (WT) mice but not in Irf5−/− mice. After whole-body inhalation exposure of CS, WT and Irf5−/− mice were decreased body wight and showed changed pressure-volume loop. In addition, the number of immune cells in the bronchoalveolar lavage fluid (BALF) was remarkably increased both in WT and in Irf5−/− mice after smoking compared to the non-smoking groups of WT and Irf5−/− mice. The CD11b+F4/80-CCR2+Ly6C+ cells were significantly increased in the BALF of smoking-exposed Irf5−/− mice compared to in that of smoking-exposed WT mice. Also, fecal microbiota analysis showed differences in bacterial abundances between WT smoking and Irf5−/− smoking mice. Taken together, our present findings suggest smoking-exposed Irf5−/− mice showed higher level of Ly6C expression in immune cells of the BALF and differential bacterial abundances in the feces. Further studies are required to elucidate IRF5-regulating Ly6C expression and determine potential therapeutic applications for COPD.
Reciprocity between IL-4 and opioid immune regulation: IL-4 primes human CD4+ T cells for opioid receptor driven MAP kinase signaling
Authors: Golovan Kenneth S , Womelsdorf Michael , Levine Alan D 
Published: August 2023

Abstract
Opioid receptor (OR) signaling induces transcriptomic and functional changes in human T cell responses at both a molecular and cellular level, yet the signaling pathways initiated by OR engagement in human T cells are poorly characterized. ORs are G-protein coupled receptors with three subtypes: MuOR, KappaOR, and DeltaOR. Using the human lymphoma Jurkat T cell line and negatively selected human peripheral blood CD4+ T cells, the effects of opioid exposure on the activation kinetics of the MAP kinases ERK 1/2, JNK, and p38, scored as their level of phosphorylation, was followed by immunoblot. Opioid-driven phosphorylation of the MAPKs was dependent on IL-4 pretreatment of Jurkat cells, but not blood-derived CD4+ T cells. After Jurkat cells were primed with a 3-day exposure to IL-4, a series of opioid agonists selective for each OR subtype induced the phosphorylation of all three MAPKs (ERK, JNK, p38) within 2 min. Phosphorylation of the MAPKs was independent of OR subtype and opioid agonist. Notably, the kinetics were different than that observed in neurons. When primary human CD4+ T cells were treated with opioids, only p38 MAPK, not ERK or JNK, was activated with similar kinetics and OR subtype independence as that seen in Jurkat cells. This difference between primary cells and Jurkat cells suggests an additional level of regulation of opioid signaling in primary cells not found in Jurkat cells. The requirement for IL-4 pretreatment suggests that opioids may play a role in Th2 biology. Preliminary data indicate that morphine modestly increases the expression of IL-4 mRNA. Overall, these results reveal that p38 functions as a primary effector of opioid receptor signal transduction in human T cells.
Supported by grants from NIH / NIDA (R01 DA 043253,)
Assessment of The Impact of Oncolytic Virotherapy on The Immunopeptidome Profile of Tumor by Utilizing GEMM Derived Immunocompetent GBM Models
Authors: Piranlioglu Raziye , Ahn Ryuhjin , Nakashima Hiroshi , Islam Anan , Guda Rahul , Charest Al , Goins William F , White Forest M , Chiocca E Antonio A 
Published: August 2023

Abstract
Immunotherapy clinical trials have not shown efficacy in glioblastoma (GBM), arguably the deadliest of all cancers. A microenvironment, characterized by a paucity of T cells recognizing tumor peptides displayed on major histocompatibility complexes of tumor cells is one of the main reasons for GBM’s evasion. Virotherapy provides the opportunity to reshape the GBM TME towards pro-inflammatory phenotype. Data from a recent clinical trial with herpes simplex-1 based oncolytic virus (oHSV) have shown that the administration of these agents leads to a rapid influx of CD8 T cells, and activation of myeloid cells, suggesting that oHSV may offer an in-situ vaccination approach to increase availability of actionable antigens. C57BL/6-derived GBM models do not mimic the outcome observed with patient derived tumors in terms of virus replication. To decipher the impact of oHSV on tumor antigen specific responses, better immunocompetent models are needed.
To assess whether oHSV treatment modulates the immunopeptidome profile (IP) of tumor, we used two transplantable GEMM-derived GBM models, 1620 and 1694. Both have wild type human EGFR, loss of Pten and Cdkn2a in BALB/c background. These models were more receptive to infection than CT2A. In vivo, virus persisted in tumors for up to 6 days, but not for 4 days in CT2A. IP analyses revealed that oHSV promotes epitope presentation on tumor cells and in TME. Moreover, low dose of virus efficiently induced antigen availability in both models though it differentially modulated the MHC ligandome.
Using two new GBM models, we showed that an oncolytic virus used in clinical trial with recurrent GBM (NCT03152318) modulates IP of tumors, and low dose of virus is sufficient to induce the presentation of tumor epitopes.
This project is supported by Bridge Fund given to Drs. Chiocca and White and P01 CA236749 to Dr. Chiocca.
Innate and adaptive mechanisms induced by BCG immunotherapy in melanoma murine model
Authors: de Queiroz Nina Marí Gual Pimenta , Borges Vinícius Martins , Caldeira Christiane Vieira Alves , Marinho Fábio Antônio Vitarelli , Oliveira Sergio Costa 
Published: August 2023

Abstract
BCG (Bacillus Calmette-Guérin) vaccine has been proposed in clinical trials for the treatment of melanoma since 1970, however its mechanism of action still not fully understood. The purpose of this study was to investigate the immune mechanisms involved in the BCG immunotherapy against melanoma. We first characterized the immunological profile of the mouse melanoma B16–F10 cell line compared to BMDMs (bone marrow-derived macrophages) infected with BCG or stimulated with agonists for different innate immune pathways. B16–F10 were only capable to respond after poly I:C and dsDNA90 stimulus inducing CXCL10 and IFN-β, but no response against BCG infection. Moreover, we evaluated BCG intratumoral treatment in a B16–F10 subcutaneous mouse tumor model in C57BL/6 wild type (WT) and several knockout mice (MyD88−/−, TLR3−/−, TLR4−/−, TLR7−/−, TLR3/7/9−/−, cGAS−/−, STING−/−, IFNAR−/−, Caspase 1−/−, Caspase 11−/− and IL-1R−/−). BCG dramatically reduces melanoma in all the mice strains, except for MyD88−/−, suggesting a role played by this adaptor molecule related to the innate immune response. In order to investigate the adaptive immunity, we evaluate BCG treatment in IFNγ−/− and Rag−/−, which demonstrated the importance of lymphocytes for tumor regression. The experiments using B16–F10 infected with BCG and co-cultured with spleen cells or BMDMs from WT or MyD88−/− mice demonstrated that MyD88 is essential for inflammatory cytokines production. MyD88 is also important to regulate the recruitment of immune cells in the tumor microenvironment. Our data suggests that BCG immunotherapy in melanoma depends on MyD88-related innate immune pathway and adaptive response for an efficient tumor control.
This study was funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPQ (152478/2022-1) and Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG (5.18/2022).
Restoration of CD4+ T helper cells without modulating the hepatic inflammatory pattern is not sufficient to prevent HCC
Authors: Manjili Masoud H , Isbell Madison , Koelsch Nicholas James , Mirshahi Faridoddin , Guo Chubquing , Idowu Michael , Austin Dana , Gelber Cohava , Wang Xiang-Yang Shawn , Sanyal Arun 
Published: August 2023

Abstract
Recent studies suggest that progression of nonalcoholic fatty liver disease (NAFLD) to hepatocellular carcinoma (HCC) is associated with a decreased frequency of the hepatic CD4+ T cells, as well as a predominance of the inflammatory immunological pattern. To determine whether the inhibition of chronic inflammation could prevent the progression of NAFLD to HCC, we used a novel LRP-1 agonistic peptide, SP16, during a progressive NAFLD in an animal model of NAFLD, i.e., DIAMOND mice. Tumor progression, circulating inflammatory cytokines as well as the hepatic immune cells were analyzed using 44-plex cytokine array and multi-color flow cytometry. Although SP16 restored the hepatic CD4+ T cells and slightly increased Th2 subset, it failed to modulate the CD4+ Th1 dominant pattern in the liver or prevent HCC. These data suggest that the hepatic immune pattern, which could produce a collective function independent from its cellular components, is more important than CD4+ T cell recovery during disease progression. Our data also suggest that administration of SP16 very early during NAFLD could shift a predominant Th1 pattern towards an equilibrium Th1=Th2=Th17 pattern which has been found to protect DIAMOND mice from from the progression of NAFLD to HCC. Future studies will determine if modulation of the inflammatory immune response by SP16 when administered early during NAFLD progression will prevent HCC.
This work was supported by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program under Award No. W81XWH2210793, Massey Cancer Center Multi-Investigator Award, grant number 2017-MIP-02, NIH R01DK105961, and VA Merit Award 1I01BX003275. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the U.S. Department of Defense. Services and products in support of the research project were generated by the Virginia Commonwealth University Flow Cytometry Shared Resource, supported, in part, with funding from NIH-NCI Cancer Center Support Grant P30 CA016059.
High-throughput method to analyze the cytotoxicity of CAR T Cells in a 3D tumor spheroid model using image cytometry
Authors: Patel Samir , Zurowski David , Hui Daniel , Ka Mignane , Hernandez Charles , Love Andrea , Lin Bo , Moore Andrea , Chan Leo Li-Ying 
Published: August 2023

Abstract
Chimeric antigen receptor (CAR)-T cell therapy is an antigen-dependent cellular therapy that has gained considerable traction in the field of cancer immunotherapy. Currently, there are six FDA-approved CAR-T cell therapies, which all target the CD19 or BCMA antigens for hematologic B cell malignancies. In recent years, a strong focus has been placed on CAR T cell therapy discovery for solid tumors, which may better recapitulate physiological conditions, thereby potentially improving the selection of CAR construct candidates. Therefore, it is critical to develop novel methodologies for high-throughput phenotypic and functional assays using 3D tumor spheroid models to better assess CAR-T cell therapies against solid tumors. Recently, plate-based image cytometry has emerged as a method to investigate and characterize CAR T cell functions in a high-throughput manner. Image cytometry has demonstrated capabilities in analyzing transduction efficiency, cell proliferation, and cytotoxicity for CAR T cell therapy. With the development of 3D spheroid models, image cytometry may provide the necessary tools and applications for CAR T cell therapy discovery geared towards solid tumors. In this work, we discuss the use of CAR-T cells targeted towards PSMA, an antigen that is found on prostate cancer tumor cells, the second most common cause of cancer deaths among men worldwide. Herein, we demonstrate the use of high-throughput image cytometry to characterize PSMA CAR-T cell-mediated cytotoxic potency against 3D prostate tumor spheroids and simultaneously monitor location of the T cells in vitro. The proposed method can effectively assess cell-mediated 3D tumor spheroid cytotoxicity and generate time- and dose-dependent results.
Identifying next generation Ig selective cleaving enzymes for treatment of autoimmune diseases using IMPACT platform
Authors: Mande Purvi , Hoyt Emily , Shin June , Pellerin Alex , Le Nam , Bengeri Soumya , Anderson Jordan , Hoel Chris , Higginson-Scott Nathan , Otipoby Kevin , Mascanfroni Ivan 
Published: August 2023

Abstract
Dysregulated humoral immune mechanisms result in immunoglobulin production that is pathogenic and contributes to a wide range of autoimmune diseases and transplant rejection. Proteases derived from human pathogens can specifically cleave these immunoglobulins (Ig) and can potentially eliminate circulating, cell-surface and immune-complex Ig classes and modulate Ig-mediated autoimmunity and inflammation, providing a novel opportunity to treat antibody-mediated diseases.
We describe here the discovery, generation, and functional characterization of Ig specific proteases. These proteases were identified using Seismic Therapeutic’s IMPACT platform and tested for their ability to cleave intact Ig subclasses and Ig isotypes in a high-throughput CE-SDS cleavage assay. Additionally, we found that these enzymes cleave soluble and cell bound Ig in human blood, dramatically reducing effector function.
We believe that these subclass and isotype specific cleaving proteases will provide an opportunity to develop novel targeted therapies for various autoimmune diseases where autoantibodies are the key drivers of the disease. In addition, we believe that they will be potentially better tolerated compared to current therapeutic interventions such as plasma exchange and other immunosuppressive therapies.
The role of M1 to M2 macrophage polarization in the etiology of idiopathic gastroparesis: GWAS perspective
Authors: Smieszek Sandra P 
Published: August 2023

Abstract
Gastroparesis is a serious medical condition characterized by delayed gastric emptying. An innate immune dysregulation and injury to the interstitial cells of Cajal and other components of the enteric nervous system is likely central to the pathogenesis of gastroparesis. We have done a GWAS comparing idiopathic and diabetic cases with controls as well as contrasting idiopathic versus diabetic cases.
We report a novel genetic association of SLC15A4 locus with idiopathic gastroparesis. This signal is driven by multiple variants with top missense variant SLC15A4:NM145648:exon2:c.T716C:p.V239A, rs33990080. Altogether, among EUR cohort we report 69 carriers out of 214 versus controls 165 out of 896 (MAF: cases 0.18; MAF: controls 0.09) and OR: 1.9 (p-value logistic model 10^−6). The significant effect persists when idiopathic cases are compared with diabetic cases as the MAF of diabetic cases (0.10) is comparable to that of population controls (0.09).
Solute carrier family 15 (SLC15) A4 is a lysosome-resident amino acid/oligopeptide transporter that is preferentially expressed in immune cells. It is required for TLR7/9-dependent type I interferon production and plays a critical role in the pathogenesis of lupus and colitis. SLC15A4 was shown to mediate M1-prone metabolic shifts in macrophages and guards immune cells from metabolic stress. Previous reports have shown that when the anti-inflammatory M2 macrophages are switched to pro-inflammatory M1 macrophages, delayed GE was evident in animal models. Current genetic findings suggest that a mechanism directly connected to macrophage polarization may be implicated in a subset of idiopathic gastroparesis patients.