bluuebunny/crossref_metadata_2025_split huggingface dataset:
A high Bacteroides/Clostridium ratio is associated with the ability of male gut microbiota transplants to suppress lupus in female BWF1 mice
Authors: Harder James W , Ma Jing , Alard Pascale , Kosiewicz Michele M 
Published: February 2023

Abstract
Systemic lupus erythematosus (SLE) is associated with microbiota dysbiosis. Our experiments conducted from 2014–16 using NZBxNZW F1 (BWF1) mice showed that gut microbiota composition differed between adult female and male mice, and male cecal transplants into female mice suppressed disease. In 2017, our BWF1 colony was moved to a new campus facility with identical water, food and bedding. Although disease incidence/kinetics were similar between the two facilities, male cecal transplants could not suppress disease in female recipients in the new facility. Several changes were made in 2018, including water source, rooms in the facility and disinfecting procedures. Transplant experiments conducted after the changes (2019–2020) recapitulated the results from our original animal facility, i.e., male cecal transplants could suppress disease in female mice. Microbiota composition in female and male BWF1 mice was compared for the three different periods. We found that Bacteroides abundance was high in male mice, and Clostridium was very low during the periods when male cecal transplants suppressed disease. Alistipes was also present during periods when male cecal transplants worked, and absent when they did not work. These same patterns were also found in females receiving male cecal transplants, with a high Bacteroides/Clostridium ratio and Alistipes presence correlating with disease suppression. These data suggest that Bacteroides and Clostridium may have opposing effects on disease suppression that may also require the presence of Alistipes. Our findings further highlight how changes in environmental conditions can have dramatic effects on microbiota composition that can directly influence the immune response and disease.
Orally-delivered microbial extracellular vesicle induces anti-inflammatory activity in mice
Authors: Argueta Shannon , Cartwright Adam , Ramani Kritika , Cormack Taylor , Chernac Fabian Romano , Hilliard-Barth Kristie , Alami Aula , Raghunathan Divya , Jalan Mihika , Caffry Will , Keats Jake , Invally Krutika , Wang Bin , Kravitz Valeria , Rommel Tyler , Ganguly Tanmoy , Ponichtera Holly , Bodmer Mark , Itano Andrea 
Published: February 2023

Abstract
The small intestinal axis is a network of anatomical and functional connections linking the small intestine with the rest of the body. It senses external signals in the gut lumen and translates them into systemic immune effects. We have previously shown that an oral microbial drug candidate induces anti-inflammatory activity in preclinical models of inflammation by acting directly on host cells without colonization of the gut or modulation of the microbiome. We now extend these observations to EDP2939, a bacterial extracellular vesicle (EV), that has potent anti-inflammatory activity in preclinical models. EVs are non-replicating bacterial membrane vesicles with approximately 1/1000th the volume of the parent cell. EDP2939 was orally delivered and gut-restricted in distribution, it acts by modulation of innate and adaptive immunity within the small intestine to attenuate systemic inflammatory responses.
We observed significantly decreased ear swelling and inflammation in a delayed-type hypersensitivity model, showing that EDP2939 modulates systemic inflammatory responses. Activity of EDP2939 is dependent upon both TLR2 signaling and the presence of local immune cells. In vitro results show TLR2 agonism and induction of anti-inflammatory cytokine responses in immune cells by EVs. This is the first report of striking anti-inflammatory effects of an orally delivered microbial extracellular vesicle. EDP2939 induces broad-based resolution of inflammation across multiple pathways via a novel mechanism of systemic pharmacology without systemic exposure. EVs are particularly effective at engaging host cells in the gut to modulate distal inflammation. These data point to oral EVs as a new class of immunotherapeutic drugs.
Treatment of Autoimmune and Inflammatory Skin Diseases Using Skin-Targeting Bifunctional Antibodies: A Localized Immunomodulation Approach
Authors: Mande Purvi , Borthakur Susmita , Rios Daniel , Halvey Patrick , Boisvert Angela , Rowe Michael , Agrawal Anisha , Cianci Mike , Borah Minasri , Viney Joanne L , Kis-Toh Katalin , Higginson-Scott Nathan , Otipoby Kevin L , Mascanfroni Ivan 
Published: February 2023

Abstract
Current treatment approaches for autoimmune conditions comprise primarily of systemic immunosuppressants or cytokine blockade. Concentration of therapeutic molecules to the tissues that are the sites of autoimmune and inflammatory diseases is a promising approach with the potential to induce therapeutic benefit and avert risks associated with systemic immunotherapies.
Pandion Therapeutics is developing a bifunctional antibody platform that can drive localized immune modulation. The platform combines a “tether antibody” that targets a tissue of choice and “an effector end” that activates specific regulatory immune pathways to restore immune-homeostasis.
Here we report the engineering of a skin-tethered PD-1 agonist and a skin-tethered CD39 that inhibit T cell activation and function and deplete local ATP, respectively, modulating different arms of the immune system in a tissue specific manner. These skin-tethered immune effectors were assessed for drug-like properties in biophysical assays and in vitro and in vivo assays for target binding, cellular activity and tissue specific-localization. Moreover, these bifunctionals were tested in pathway-relevant preclinical models such as Vitiligo and Contact Hypersensitivity. Strikingly, a selective accumulation of the tethered bifunctionals to the skin was observed and correlated with a tether-dependent efficacy compared to a non-tether control.
We believe that this therapeutic approach has the potential to drive the resolution of cutaneous inflammation, providing an opportunity for developing new targeted therapies for autoimmune and inflammatory skin diseases.
Alphataxin, a Small-molecule Drug that Elevates Tumor-infiltrating CD4+ T Cells, in Combination with Anti-PD-1 Therapy, Suppresses Murine Renal Cancer and Metastasis
Authors: Bristow Cynthia L , Reeves Mary Ann B. , Winston Ronald 
Published: February 2023

Abstract
By promoting the cytotoxic function of CD8+ T cells, immune checkpoint inhibitor therapy, e.g. programmed cell death protein-1 (PD-1), effectively inhibits tumor growth in renal cell carcinoma. Yet, as many as 87% of cancer patients do not respond to immune checkpoint therapy. The function of cytotoxic CD8+ T cells integrally relies on CD4+ T helper cells. Remarkably, despite advances in immunotherapy, there are no pharmaceutical treatments that increase circulating CD4+ T cell counts. Nor has there been much attention given to tumor-infiltrating CD4+ T cells. The orally available small-molecule drug Alphataxin targets a pathway that regulates the number of circulating CD4+ T cells. We aimed to examine how Alphataxin affected tumor growth in a murine model of renal cell carcinoma. Alphataxin, in combination with anti-PD-1 antibody, significantly elevated the ratio of circulating and tumor-infiltrating CD4+/CD8+ T cells. In one study, following orthotopic implantation of syngeneic renal cell carcinoma, combination treatment resulted in 100% regression of tumor growth. Moreover, in mice implanted orthotopically with one log more tumor cells, doubling Alphataxin dose in combination treatment led to 100% regression in one-third of mice and 81% suppression of tumor growth in the remaining two-thirds of mice. Lung metastasis was evident in all mice except combination-treated mice. Orally available Alphataxin, the first and only drug developed to increase CD4+ T cells, in combination with anti-PD-1, is a powerful therapeutic method that provides long-term remission in renal cell carcinoma and potentially other T cell-responsive cancers by increasing the number of CD4+ tumor-infiltrating T cells.
Irreversible electroporation stimulates a pro-inflammatory tumor microenvironment that facilitate antigen presentation in pancreatic cancer
Authors: Imran Khan Mohammad , Brock Rebecca M. , Beitel-White Natalie , Hendricks-Wenger Alissa , Coutermarsh-Ott Sheryl L , Manuchehrabadi Navid , Davalos Rafael V. , Allen Irving C 
Published: February 2023

Abstract
Pancreatic cancer is now the third leading cause of cancer-related death and is projected to turn out to be the second by 2030. Pancreatic cancer is often late-diagnosed with few treatment options with extremely low survival rates. Surgical candidacy is only 15% of total diagnosed patients. Irreversible electroporation (IRE), a novel form of tumor non-thermal ablation applies high-voltage microsecond pulses to permeabilize the cell membrane that is irreversible and leads to apoptotic cell death. IRE has had strong success in clinical trials for pancreatic cancer patients. Nevertheless, little is known on the effect of IRE beyond the initiation of cell death in the tumor tissue. We hypothesize that IRE can induce a pro-inflammatory type of cell death in pancreatic cancer that can promote inflammation and anti-tumor immune system responses. We have used in vitro, ex vivo, and immunocompetent in vivo murine models to investigate alterations to pancreatic cancer and its microenvironment. Our findings reveal programmed necrotic cell death, promotion of pro-inflammatory cytokine production, and shifts in immune cell populations from a pro-tumor (such as MDSCs and T regulatory cells) to a proinflammatory types containing cytotoxic lymphocytes and neutrophils after IRE treatment. Progression-free survival was significantly longer for the treated mice. Intriguingly, IRE treatment increases IFNγ signaling and generates viable antigens for presentation to the adaptive immune system but also promotes PD-L1 expression which in turn causes immunological stalemate. Our results suggest possible therapeutic targets for use in combination with IRE, such as gemcitabine or PD-1/PD-L1 inhibitors, to improve patient survival.
Oral delivery of a microbial extracellular vesicle induces potent anti-tumor immunity in mice
Authors: Francisco Loise , Abdou Mary , Troy Erin , Ballok Alicia , Chernac Fabian Romano , Kashyap Shubhra , Argueta Shannon , Tsang Jessica , Goldberg Michael , Sizova Maria , McBride Audrey , Keats Jake , Invally Krutika , Rommel Tyler , Ponichtera Holly , Huynh Kevin , Kravitz Valeria , Ganguly Tanmoy , Bodmer Mark 
Published: February 2023

Abstract
The small intestinal axis (SINTAX) is a network of anatomic and functional connections between the small intestine and the rest of the body. It acts as an immunosurveillance system, integrating signals from the environment that affect physiological processes throughout the body. The impact of events in the gut in the control of tumor immunity is beginning to be appreciated. We have previously shown that an orally delivered single strain of commensal bacteria induces anti-tumor immunity via activation of innate and adaptive immunity. Some bacteria produce extracellular vesicles (EVs) that share molecular content with the parent bacterium in a particle that is roughly 1/1000th the volume in a non-replicating form. We report here an orally-delivered and gut-restricted bacterial EV which potently attenuates tumor growth to a greater extent than whole bacteria or checkpoint inhibition. EDP1908 is a preparation of EVs produced by a bacterium of the Oscillospiraceae family isolated from a human donor. Treatment of tumor-bearing mice with EDP1908 shows superior control of tumor growth compared to anti-PD-1 or an intact microbe. EDP1908 significantly increases the percentage of IFNγ producing CD8+, NK, NKT and CD4+ cells in the tumor microenvironment (TME). EDP1908 also increases tumor-infiltrating dendritic cells (DC1 and DC2) and production of IP-10 and IFNg in the TME, creating an environment conducive to the recruitment and activation of anti-tumor lymphocytes. These data point to oral EVs as a new class of immunotherapeutic drugs that harness the biology of the small intestinal axis, acting locally on host cells in the gut to control distal immune responses within the TME. EDP1908 is in preclinical development for the treatment of cancer.
‘MR1 Tetramer-based Artificial Antigen Presenting Cells Expand MAIT Cells from Human Peripheral Blood that Effectively Kill Glioblastoma Cells.’
Authors: PRIYA RAJ , BRUTKIEWICZ RANDY RICHARD 
Published: February 2023

Abstract
Immunotherapy for cancer treatment requires the activation of cytotoxic effector lymphocytes. Mucosal-associated invariant T (MAIT) cells are innate T cells that recognize the MHC class I-like molecule, MR1. MAIT cells play an important role in the immune response against microbial infections and can directly kill tumor cells. Although MAIT cells can be expanded ex vivo, this method is time-consuming, expensive and requires allogenic feeder layers. To overcome the limitations of conventional dendritic cell-based vaccines and ex vivo expansion of human T cells, an artificial antigen presenting cell (aAPC) approach to expand antitumor effector cells has several advantages. In this study, we explored an efficient in vitro method to amplify MR1-specific MAIT cells from human peripheral blood using aAPCs made by coating cell-sized latex beads with an antigen-loaded MR1 tetramer complex and anti-CD28 antibody. We further elucidated the cytotoxic potential of such expanded MAIT cells against three human glioblastoma (GBM) cell lines to explore their potential use as a novel immunotherapeutic tool, as the mostly lethal GBM poorly responds to conventional chemotherapy. When aAPCs were compared to the standard allogenic feeder layer-based approach for MAIT cell expansion, they were significantly more effective. Our results indicate that the aAPC-expanded MAIT cells remained functional, retained their original phenotype, secrete pro-inflammatory cytokines, and showed cytotoxicity against the GBM cell lines. Hence, MAIT cells have the potential to be a novel tool in immunotherapy approaches for the treatment of human GBM.
Seq-ing the source of radiation-induced anti-tumor immunogenicity in patient tumors
Authors: Chow Jacky Louis , Hoffend Nicholas , Gaudieri Madeline , Abrams Scott I , Schwaab Thomas , Singh Anurag , Muhitch Jason 
Published: February 2023

Abstract
Translational studies in a small number of patients have demonstrated a clinical benefit when combining radiation and immunotherapy. These findings are supported by correlative data showing that transcriptional changes to the tumor microenvironment following radiotherapy are associated with responses. Yet, given limited access to patient specimens following treatment, few studies have addressed how radiation primes anti-tumor immunity at single cell resolution, thus limiting our understanding of the source of radiation-induced changes within tumors. Here, we acquired renal cell carcinoma (RCC) tumors from patients treated with stereotactic body radiation therapy (SBRT) followed by nephrectomy in a pilot study (NCT01892930), to evaluate radiation-induced transcriptional changes to immune and RCC cells by single-cell RNA sequencing. We hypothesized that SBRT-induced interferon (IFN) and immune-activating signals, previously detected by bulk sequencing and array platforms, would localize to the dense lymphocyte infiltrate characterized in RCC and that the transcriptional signatures in SBRT-treated RCC would reflect increased immune recognition. Single-cell transcriptomics uncovered increased frequency of mature T cells in irradiated tumors, including increased expression of IFN-gamma and granzymes, as well as PD1 and other exhaustion markers. Additionally, irradiated-RCC cells expressed more IFN-gamma receptor and were enriched for IFN signaling and antigen processing pathways. Collectively, these results indicate radiation may improve anti-tumor immunity by increasing both T cell activity and tumor cell sensitivity to pro-inflammatory cytokines and cytolysis in human malignancies.
Quantitative Immune Profiling of Human Tumor Tissues with Multiplexed ChipCytometry
Authors: Gamber Kevin , Christians Arne , Schwarz Spencer , Northcutt Adam , Forys Jason , Campbell Thomas , Winkeler Crystal 
Published: February 2023

Abstract
Cancer therapies that rely on manipulating or engineering immune cells have shown much promise in recent years for effectively combating a broad array of cancer types. In order to determine the effectiveness of such therapies, it is necessary to accurately profile the complement of immune cell types present in the tumor microenvironment. Methods to reliably obtain immune cell signatures require the combination of powerful resolution to discriminate cell boundaries, broad dynamic range to capture markers of varying expression levels, and accurate cell segmentation across a wide range of cell sizes and morphologies. Many methods for quantitative immune cell profiling have demonstrated shortcomings when it comes to the aforementioned attributes. To remedy these shortcomings, we present here ChipCytometry, an imaging technology for immune profiling of cells and sectioned tissues.
ChipCytometry is a fluorescence-based imaging system that utilizes multiplexed immuno-fluorescence staining in combination with high-dynamic-range (HDR) imaging to facilitate quantitative phenotyping of individual cells within tissue samples. Employing this technology to profile metastatic and primary tumors, multiple biomarkers were stained in iterative multiplex assays to profile single cells with an AI-powered cell segmentation algorithm. The HDR imaging allows for both strong and weak fluorescent signals to be simultaneously obtained without loss of sensitivity. This immune profiling was completed on 6 different cryosectioned human tumor tissues (head & neck, liver, lung, breast, colon, and pancreas).
Circulating immune cell phenotype dynamics reflect the effect of a CD200 immune checkpoint targeted immunotherapy in pet dogs with high-grade glioma
Authors: Puerta-Martinez Francisco W , Schnell Sathya , Arnold Susan , Pluhar G. Elizabeth , Olin Michael 
Published: February 2023

Abstract
The extent to which immune cell phenotypes in the peripheral blood reflect intratumoral immune activity prior to and early in cancer immunotherapy is unclear. Here we studied the population dynamics of immune cells and immune phenotypic changes, using clinical blood samples and a multicolor flow cytometry analysis. These samples were serially obtained from a cohort of high-grade glioma canine patients enrolled in a trial with an immunotherapy that targets the CD200/CD200R immunological synapse. This therapy consists of an alternative checkpoint inhibitor peptide (CD200AR-L) that modulates the CD200 immune checkpoint, currently being used in an adult phase I safety trial for recurrent glioblastoma trial (NCT04642937) and a canine high-grade spontaneous glioma trial (U01CA224160-01). We found evidence of a strong effect of the therapy on peripheral immune cells in responder versus nonresponder dogs. Upon initiation of therapy, immune effector cell proliferation increased in responsive patients, whereas in nonresponders immune cell populations either didn’t change significantly or an increase in immunosuppressive phenotypes was observed. Cell populations such as MDSC, regulatory T cells and exhausted T cells were predominantly higher in nonresponder dogs. These data showed that peripheral immune cell phenotypes were linked to the outcome of the immunotherapy, thus dogs with better immune cell profiles, including increased cytotoxic differentiation, had a better prognosis and longer overall survival rates. Taken together, these results suggest that variation in patient responsiveness can be explained by differences in peripheral immune cell proliferation and differentiation soon after the initiation of immunotherapy.
STAT1 associated immune checkpoint expression and immune cell localization represent adaptive immune resistance in high-grade serous ovarian cancer
Authors: Li Deyang , Vidotto Thiago , Shakfa Noor , Afriyie-Asante Afrakoma , Peterson Nichole , de Ladurantaye Manon , Mes-Masson Anne-Marie , Francis Julie-Ann , Koti Madhuri 
Published: February 2023

Abstract
High-grade serous carcinoma (HGSC) is a deadly malignancy leading to ~70% of the 140,000 ovarian cancer deaths each year. Resistance to platinum chemotherapy followed by recurrence and an incurable disease occurs in most HGSC patients. Contemporary immune checkpoint blockade therapies have shown minimal efficacy in this cancer. Our previous investigations established that tumour interferon (IFN) activation status and CD8+ T cell density are predictors of chemotherapy response in HGSC. Furthermore, we also showed that the IFN induced chemokine CXCL10 is a key determinant of increased survival via immune cell recruitment in the tumour immune microenvironment (TIME). Given that signal transducer and activator of transcription 1 (STAT1) is central to the feed forward loops of cellular IFN responses, we investigated STAT1 associated transcriptomic alterations and spatial profiles of immune cells in 204 pre-treatment HGSC tumours.
RNA-sequencing based whole transcriptomic profiling revealed that higher STAT1 expression significantly correlated with higher immunomodulatory gene expression, including immune checkpoints and activators, in both chemotherapy sensitive and resistant tumours. Findings were independently validated in a cohort of 379 HGSC tumour RNA-Seq profiles from The Cancer Genome Atlas Network ovarian cancer dataset. Multiplex immunofluorescence based spatial profiling of CD8+ T cells, FoxP3+ T regulatory T cells, CD68+M1, and CD163+ M2 macrophages and expression of PD-L1, PD-1, IDO1 immune checkpoints was performed. Findings from our study provide evidence for IFN mediated adaptive immune resistance in the HGSC TIME and will potentially inform the design of rational chemo-immunotherapy approaches.
Induction of Innate Immune Visibility of Neuroendocrine Tumors Potentiates Immune Responses and Tumor Rejection
Authors: Zhu Mingrui , Huang Yi , Bender Matthew , Girard Luc , Kollipara Rahul , Eglenen-Polat Buse , Naito Yujiro , Savage Trisha , Huffman Kenneth , Koyama Shohei , Kumanogoh Atsushi , Minna John , Johnson Jane , Akbay Esra 
Published: February 2023

Abstract
Natural killer (NK) cells serve as an important role in suppressing tumor growth by directly attacking malignant cells. These cells recognize the tumor cells through moleclar patterns associated with oncogenic transformation. NKG2D ligands (NKG2DLs) are one of these markers present on tumor cells which can activate NK cells and T cells by interacting with NKG2D receptors on these lymphocytes. Majority of tumors express NKG2DLs. Interestingly, tumors carrying neuroendocrine features such as small cell lung cancer (SCLC) and neuroblastoma express very low level of ligands for NK cell activating receptor NKG2D (MICA, MICB, ULBP1, ULBP2, ULBP3), which may help them escape from innate immune surveillance. Further characterization of SCLC immune environment showed reduced antigen presentation, reduced infiltration of total immune cells specifically NK cells, as compared to NSCLC, a more immunogenic tumor. Transcriptional inducers of NKG2DLs did not induce NKG2DL in SCLCs due to hypo-acetylation of MICA/B region. Histone deacetylase (HDAC) inhibitors were able to induce NKG2DL on tumor cell surface. Restoring NKG2DL in preclinical SCLC and neuroblastoma models either by ectopic expression or HDAC inhibitors stimulated direct NK cell killing in co-culture assays. NKG2DL stimulation also caused potentiation of antibody directed therapies such as CD3-EpCAM in neuroendocrine tumors. In vivo studies showed suppressed tumor growth and metastasis in an NK and CD8 T cell dependent manner when NKG2DLs are stimulated. We conclude that: restoring innate immune visibility can target neuroendocrine tumors to both innate and adaptive immune cytotoxicity.