bluuebunny/crossref_metadata_2025_split huggingface dataset:
Type-1 Interferons impair the immunoregulatory activity of IL-10: understanding novel mechanisms of abrogation of transplant tolerance
Authors: Lozano Marcos Iglesias , Bibicheff Darrel , Chicco Maria , Brandacher Gerald , Raimondi Giorgio 
Published: January 2023

Abstract
Induction of long term transplant survival by “costimulation blockade” (CoB) regimens is impaired by inflammatory responses. Type-1 interferons (TI-IFNs) are recognized mediators of this effect in multiple models, but their target population and specific mechanism used remain undefined. To better understand how TI-IFN could interfere with transplant tolerance, we studied their impact on the immunomodulatory properties of IL-10.
Using a murine full-mismatch skin transplant model we showed that IL-10 has a fundamental role in the protective effect of CoB. Bone marrow chimeras suggested this effect is partially dependent on IL-10 signaling in T cells. We then studied the impact of T cell exposure to TI-IFN on IL-10 signaling. Following in vitro exposure, T cells produced significantly less phospho-STAT3 in response to IL-10. This effect was very selective - IL-6 signaling remained unaltered - and resulted in the inhibition of specific IL-10 induced genes. Encouragingly, this effect was slowly reversible with removal of TI-IFN, suggesting a “druggable” pathway. This unresponsiveness was not associated with reduced IL-10-receptor expression or increased SOCS1_3 levels. Instead, microarray analysis of exposed cells suggested a novel role for STAT1 in dampening IL-10 signaling. Preliminary experiments with STAT1-KO cells supported this hypothesis.
Overall, these results highlight the importance of IL-10 in the efficacy of CoB regimens and reveal a new molecular mechanism whereby TI-IFNs can interfere with IL-10 signaling, and ultimately with the regulation of alloreactivity. A strategy to selectively target this mechanism could significantly enhance the clinical efficacy of CoB-based immunomodulatory strategies.
Inhibiting EZH2 function prevents and treats chronic graft vs host disease (cGVHD) by disrupting germinal center formation and plasma cell maturation
Authors: Zaiken Michael C , Flynn Ryan , Paz Katelyn , Rhee Stephanie , Park Paul , Hemming Matthew , Sage Peter , Sharpe Arlene , Luznik Leonard , Mailard Ivan , Hill Geoffrey R. , MacDonald Kelli , Serody Jonathon , Cutler Corey , Koreth John , Miklos David , Antin Joseph , Soiffer Robert , Ritz Jerome , Bradner Jay , Qi Jun , Blazar Bruce R. 
Published: January 2023

Abstract
To determine whether targeting EZH2, a histone methyltransferase necessary for germinal center (GC) formation, could reduce murine cGVHD (cGVHD), we first used a clinically relevant murine model of cGVHD, which recapitulates multi-organ cGVHD with bronchiolitis obliterans (BOS). Bone marrow (BM) from EZH2 fl/fl CD19 Cre B6 mice and WT T cells, or WT B6 BM and EZH2 fl/fl CD4 Cre B6 T cells were given to conditioned B10.BR mice. EZH2 expression in BM and T cells was obligatory for cGVHD. To determine whether EZH2 function could be targeted to treat cGVHD, we tested a novel EZH2 small molecule inhibitor, JQ5 with a favorable toxicity and efficacy profile to other inhibitors. Mice with established cGVHD given JQ5 had dramatically improved cGVHD. Similar results were seen in a sclerodermatous cGVHD model. To assess possible causes of this improvement we performed RNA-seq and ATAC-seq data of MACS purified CD19+ B cells obtained from control and JQ5 treated BOS cGVHD mice. Unsupervised gene set enrichment analysis and weighted gene correlation network analysis showed that among gene clusters associated with JQ5 treatment there was an enrichment for genes expressed in naïve B cells compared to plasma cells (GSEA4142) and an impairment of genes associated with late stage GC B cells compared to early follicular B cells (GSEA11961). This suggested that JQ5 impaired GC B cell and plasma cell maturation. Consistent with these data, splenic follicular cells, GC size and number, and lung resident PCs were reduced compared to cGVHD controls. Taken together these results show that EZH2 is necessary for the initiation of cGVHD and show for the first time that EZH2 inhibition using a novel inhibitor JQ5 can reverse established cGVHD in mice with BOS or scleroderma.
Donor Vγ4+T cells promote graft-versus-leukemia effect and ameliorate acute graftversus- host disease in allogeneic hematopoietic stem cell transplantation
Authors: Song Yuan , Zhu Ying , Liu Haiyan 
Published: January 2023

Abstract
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is one of the most effective ways of treating leukemia and other malignant diseases. However, disease recurrence and GVHD are still the major causes of death after allo-HSCT. Previous studies showed that immune cells involved in graft versus leukemia (GVL) effect traditionally include CD8+ T, CD4+ T and NK cells. Unfortunately, some of these T-cell subsets also cause graft-versus- host disease (GVHD). The role of donor γδT cells after transplantation is less well defined. We detected direct anti-leukemia cytotoxicity of γδT cells in vitro and the cytotoxicity of Vγ1 subset was stronger than the Vγ4 subset. To evaluate the role of donor γδT cells in aGVHD and GVL, we established the murine HLA fully mismatched allo-HSCT GVL/GVHD model. Our results showed that TCRδ−/− donor bone marrow cells caused enhanced disease progression and aggravated aGVHD compared with wildtype donors. Furthermore, flow cytometry analysis demonstrated γδT cells mediated anti-leukemia effects through promoting IFN-γ production by CD8+ T cells while suppressed aGVHD by inhibiting the activation of CD4+ T cells in liver, lung and IEL. The experiments of adoptive transfer of in vitro expanded γδT/Vγ1/Vγ4 cells and the in vivo depletion of Vγ1/Vγ4 cells indicated that the subset of donor γδT cells to alleviate murine aGVHD and enhance GVL effect might be Vγ4 γδT cells. The GVL effect of Vγ4 cells in vivo could be mediated by the production of IL-17A and regulation of CD8+ T cell function. These findings suggest donor γδT cells infusion could be a novel strategy to improve patients’ overall survival after allo-HSCT.
Bcl-2 Inhibition with Venetoclax Promotes Induction of Mixed Chimerism and Renal Allograft Tolerance Without Severe Myelosuppression in Non-human Primates
Authors: Kawai Tatsuo , Sasaki Hajime , Ma David , Dehnadi Abbas , Cosimi A Benedict , Cippa Pietro , Fehr Thomas 
Published: January 2023

Abstract

Background
Specific Bcl-2 inhibition has been shown to promote mixed chimerism and allograft tolerance without myelosuppressive conditioning in murine models. We extended this approach to nonhuman primates in combined kidney and bone marrow transplantation (CKBMT).

Methods
In Group A, recipients received CKBMT with the regimen including total body irradiation (TBI,3 Gy) thymic irradiation (TI, 7Gy) and ATG, followed by a short course of anti-CD154 antibody and cyclosporine. In Group B, TBI was reduced to 1.5Gy. In Group C, peri-transplant Bcl-2 inhibitor (Venetoclax) was added to Group B regimen. In Group D, Group C regimen without TBI. In Group E, Group C regimen without TI.

Results
7/8 recipients in Group A developed transient chimerism and achieved long-term renal allograft survival without immunosuppression. However, these recipients developed transient but severe pancytopenia between days 10–17 post CKBMT. Both recipients in Group B failed to develop chimerism. By adding Venetoclax to Group B regimen, 3/3 Group C recipients developed significantly higher and longer mixed chimerism without severe myelosuppression. These three recipients also achieved long-term immunosuppression free renal allograft survival without rejection (&gt;316, &gt;575, &gt;239 days). In Group D, both recipients failed to develop chimerism and rejected their allografts on days 140 and 120. Although three Group E recipients successfully developed chimerism, all rejected their renal allografts on days 100, 97, 163 with early recovery of CD31+ naive T cells.

Conclusion
Enhancement of intrinsic apoptosis with Bcl-2 inhibition is a promising strategy to achieve robust mixed chimerism and allograft tolerance with reduced myelosuppressive conditioning.
Enhancing Antigen Delivery Using Spherical Nucleic Acidsfor Optimal Prostate Cancer Immunotherapy
Authors: Qin Lei , Wang Shuya , Dominguez Donye , Long Alan , Chen Siqi , Fan Jie , Ahn Ji Hae , Skakuj Kacper , Lee Andrew , Mirkin Chad , Zhang Bin 
Published: January 2023

Abstract
Prostate cancer ranks as the second leading cause of cancer-related death in the United States in men. For patients with metastatic or recurrent disease, cancer vaccines remain an attractive treatment approach while therapeutic efficacy is rather limited. Here, we developed Spherical Nucleic Acid nanostructures (SNAs) to deliver adjuvant CpG and prostate cancer peptide antigens to potentiate antitumor responses. We show that SNAs that differ in the positioning of the antigen within the nanostructure lead to differences in the quality of immune responses generated by vaccination with SNAs. SNAs conjugated with both antigen and adjuvant CpG on the surface (HM SNAs)induce stronger cytotoxic T lymphocyte (CTL) mediated antigen-specific killing of target cells than that for SNAs with CpG on the surface and antigen encapsulated within the core (EM SNAs). Mechanistically, HM SNAs increase co-delivery of CpG and antigen, thereby promoting dendritic cells-mediated cross-priming compared to EM SNAs, or admix of free peptide and linear CpG. Furthermore, vaccination with HM SNAs led to significantly inhibits tumor growth in two syngeneic mouse models of prostate cancer. These data suggest that the positioning of peptide antigens within SNA structures can be exploited to enhance the therapeutic performance of SNAs, representing a novel approach to vaccine design for cancer immunotherapy and potentially other diseases that can be addressed through modulating the immune system.
Assessing the impact of neoantigen load on checkpoint blockade efficacy
Authors: Yalcin Zeynep Kosaloglu , Frentzen Angela , Premlal Ashmitaa Logandha Ramamoorthy , Sidney John , Greenbaum Jason A , Sette Alessandro , Peters Bjoern 
Published: January 2023

Abstract
Neoantigens were suggested to be crucial for the outcome of immune checkpoint blockade (ICB) therapies and accordingly, the neoantigen load of a tumor is often being used to predict therapy outcome. Current approaches utilize HLA binding predictions to rank peptides, and the neoantigen load of a given tumor is then usually defined as the number of peptides binding above a certain threshold. Using this approach some reports found a strong correlation of neoantigen load and response to ICB while others found a better correlation when mutational load was used instead. However, comparing results between studies is hard because they used different tools and thresholds to identify mutations. We sought to overcome this lack of uniformity in re-analyzing data from several published studies to consistently define the mutational burden and neoantigen load, and correlate them with therapy outcomes.
We obtained sequencing data from several ICB studies and run all datasets through our in-house mapping and variant calling pipelines. We applied different scoring schemes to define the neoantigen load which were based on HLA binding predictions with varying thresholds and different metrics used. We found that the exact definition of neoantigen load has a major impact on how well this metric performs in predicting checkpoint blockade therapy outcome. We also implemented a likelihood score which is based on data of known immunogenic neoantigens and describes the likelihood of a predicted neoantigen to be immunogenic. Based on our preliminary analysis to date, we found that the cumulative likelihood score of a patient is a good predictor of ICB outcome, outperforming tumor mutational burden and predicted neoantigen load in several cohorts.
Adjuvant screen identifies DC modifying growth factor Flt3 ligand as candidate for prostate cancer DNA vaccine
Authors: Malo Kirsten , Wong Ashley J. , Thorne Amy H. , Humeau Laurent , Masteller Emma 
Published: January 2023

Abstract
Prostate cancer is the most common type of cancer in men. Immunotherapies such as Sipuleucel-T have shown that stimulating the immune system to target the prostate is a viable therapeutic option. Inovio is a clinical-stage biotechnology company that uses DNA vaccines as a novel immunotherapy strategy. An advantage to plasmid DNA vaccine therapy is the ability to encode adjuvants within the vaccine in order to increase immunogenicity and efficacy.
We conducted an adjuvant screen in BALB/c mice with 25 candidate plasmid encoded genetic adjuvants in combination with the prostate cancer specific tumor-associated antigen STEAP1. Antigen-specific T cell responses were measured by IFNg ELISpot. The screen revealed that the addition of a plasmid encoding dendritic cell (DC)-activating Fms-like tyrosine kinase 3 ligand (Flt3L) fused to an Fc domain, significantly increased antigen-specific T cells (p&lt;0.001, 2.8 fold). Mice treated with Flt3L-Fc had an enhanced immune response to STEAP1 vaccination as early as 7 days post dose 1 (p&lt;0.01, 2.4 fold), which developed into an enhanced memory response measured 12 weeks later (p&lt;0.001, 8.5 fold). Flow cytometry showed that Flt3L-Fc increased DC populations at the site of injection and at the draining lymph node 8 days following the initial vaccination. DCs are considered the most potent antigen-presenting cells in the immune system and DCs at the tumor site have been shown to be critical for T cell immunity. Our data shows that enhancing DC populations and function through the use of a genetic adjuvant can enhance the immunogenicity of a DNA cancer vaccine. Future studies will assess the efficacy of Flt3L-Fc in combination with STEAP1 in a mouse tumor model.
Regional pressure-enabled drug delivery of anti-PD-1 agent for colorectal liver metastases improves anti-tumor activity without increased hepatic toxicity
Authors: Chai Louis F. , Hardaway John C. , Lopes Mikayla C. , Rabinowitz Benjamin A. , Guha Prajna , Jaroch David , Cox Bryan , Espat N. Joseph , Katz Steven C. 
Published: January 2023

Abstract

Introduction
Checkpoint inhibitors (CI) have greatly impacted the treatment of solid tumors. Success in the liver has been more limited. When intravenously infused, the CI is delivered predominantly to extrahepatic sites, which results in systemic side effects. Previously, we reported on regional delivery of cellular immunotherapy for liver tumors using pressure-enabled drug delivery (PEDD™, TriSalus™ Life Sciences, Inc.). We hypothesized that PEDD of CI would improve the therapeutic index (TI).

Methods
In a murine model of colorectal liver metastases (CRLM), we infused high (HP) or low pressure (LP) anti-PD-1 antibodies (RMP1-14) via the portal vein and compared to systemic tail vein delivery (SD) at 0, 0.3, 1.0 and 3.0 mg/kg. Tumor bioluminescence (TB), histopathology, and serologic analysis informed treatment effects and toxicity.

Results
At a dose of 3mg/kg, LP resulted in similar IVIS TB growth kinetics compared to SD 3 mg/kg at 7 days post-treatment (geometric mean, p = 0.94). In animals treated with a 10-fold lower dose (0.3 mg/kg), tumor growth was significantly lower with HP vs. LP (7.5-fold improvement, p = 0.04). Circulating serum anti-PD-1 levels were similar when we compared SD and LP at 1 and 3 mg/kg (Mean 2512.0 vs. 2250.2 ng/mL, p = 0.25). Comparatively, systemic exposure was significantly lower at 0.3 mg/kg LP (Mean 508.1 ng/mL, p &lt; 0.0001). No increases in hepatotoxicity were seen with PEDD (AST p = 0.57, ALT p = 0.43).

Conclusion
PEDD of anti-PD-1 CI improved liver tumor control while also limiting systemic exposure. These findings support PEDD for anti-PD-1 therapy to improve TI. Phase 1 exploration of anti-PD-1 PEDD for patients with hepatic malignancy is thus warranted.
Interleukin-15 in cancer immunotherapy: IL-15 receptor complex versus soluble IL-15 in a cancer cell-delivered murine leukemia model
Authors: Berger Alexandra , Colpitts Sarah , Seabrook Melanie , Furlonger Caren , Bendix Maura , Moreau Joshua , McKillop William , Medin Jeffrey , Paige Christopher J 
Published: January 2023

Abstract
Cancer immunotherapy endeavors to stimulate the immune system in order to recognize, reject and destroy tumor cells. Previously we have demonstrated that IL-12 is an ideal candidate for leukemia immunotherapy and there is a Phase 1 human trial underway at the Princess Margaret Cancer Center to test this approach in AML patients. In order to further our knowledge on immunomodulators we have expanded our studies to other cytokines such as IL-15. IL-15, either in its soluble form (IL-15sol) or complexed with IL-15Ra (IL-15Rc), has been shown to exhibit potent anti-tumor activities in various cancer studies.
In this study we describe the impact of intraperitoneal IL-15 in a cancer cell delivered IL-15 immunotherapy approach using the 70Z/3-L leukemia mouse model. Whereas both forms of IL-15 led to significantly improved survival rates compared to the parent cell line, there were striking differences in the extent of the improved survival: mice receiving cancer cells secreting IL-15sol showed near 100% survival for up to 250 days, whereas mice receiving cancer cells secreting IL-15Rc started to perish around day 50 post-injection, with few survivors left on day 250.
Interestingly, cell-secreted IL-15sol and IL-15Rc activated different cell types in the peritoneum, leading to different immune responses. IL-15Rc resulted in an influx and/or expansion of NK1.1+ natural killer cells in the peritoneum, while injection of cells secreting IL-15sol lead to an increased expansion of CD4+ and CD8+ T-cells. To our knowledge, this is the first study detailing a significantly different biological effect of cell-delivered IL-15Rc vs IL-15sol in a mouse cancer immunotherapy study, suggesting a different mechanism of action of the two IL-15 forms.
Lenolidamide and IL-15 enhance natural killer cell-mediated immunity and augments the anti-tumor effect of an anti-CD30/CD16A bispecific antibody against Hodgkin’s lymphoma B-cells
Authors: Manna Alak , Aulakh Sonikpreet , Ailawadhi Sikander , Chanan-Khan Asher , Sher Taimur , Paulus Aneel 
Published: January 2023

Abstract
Hodgkin’s lymphoma (HL) is characterized by malignant CD30+ Reed-Sternberg (HRS) B-cells surrounded by an ineffective infiltrate of inflammatory immune cells. T-helper and T-regulatory cells presumably provide survival signals for HRS cells; rescuing them from immune-surveillance by CD8+ cytotoxic T lymphocytes and natural killer (NK) cells, which are also functionally anergic. Clinically, ≈10–30% of HL patients resulted in relapse after front line therapy; highlighting the need for development of new treatments. We investigated a novel bispecific CD16axCD30 antibody, AFM13 (Affimed Therapeutics, AG), which engages NK cells and recruits them towards CD30+ tumor cells. Safety of single-agent AFM13 has been demonstrated in a phase I clinical study, however, disease remission was observed in only 23% of patients. We hypothesized that combining AFM13 with another agent whose safety profile is already been established in HL and also activates NK cells (lenalidomide) could potentially increase anti-tumor efficacy of AFM13. We observed increased expression of CD107a, CD69, CD25 and IFNγ (activation markers) in NK cells isolated from HL patients (HL-NK), that were stimulated ex vivo with IL- 15 + lenalidomide. Pre-treatment with IL-2/IL-15 antibody has been shown to activate and restore T/NK cell cytolytic function. In line with this, IL-15+lenalidomide-stimulated HL-NK cells showed functional restoration by inducing specific lysis of human CD30+ lymphoma cells (30.55±1.49%) in a co-culture assay. While, AFM13 is being investigated in combination with anti-PD1 antibody therapy (NCT02665650), our preclinical data provides the rationale for an AFM13 + Len combination regimen for HL and potentially other CD30+ malignancies.
Towards identification of a lipid metabolic checkpoint for immuno-oncology
Authors: Hsu Ku-Lung , Borne Adam L. , Brulet Jeffrey W. , Campbell Sean T. , Franks Caroline E. , Huang Tao , McCloud Rebecca L. , Shin Myungsun , Ware Timothy B. 
Published: January 2023

Abstract
Lipid phosphorylation is an understudied regulatory mechanism for T cell metabolism and signaling. Specifically, diacylglycerol kinases (DGKs) modulate intracellular levels of the secondary messengers diacylglycerol and phosphatidic acid, which are implicated in regulation of T cell activation and anergy. Development of isoform-selective DGK inhibitors is challenging but needed to understand specificity of DGK biology in vivo. Towards this goal, we use ATP acyl phosphate activity-based probes and quantitative mass spectrometry to define, for the first time, the DGK ATP-binding site (DAGKc/DAGKa subdomains) and propose a potential role for C1 domains in DGK catalytic function. We discovered fragments from old drugs (ritanserin) that reveal pharmacophores required for selective binding to DGK active sites that are not in the canonical ATP binding pocket. We provide evidence of a single contiguous inhibitor binding site composed of DAGKc, DAGKa, and C1 sites by demonstrating equipotent inhibition using a fragment inhibitor. Our current working model is that C1 domains of DGKs are involved in substrate and inhibitor recognition of type 1, 4, and 5 DGKs and is a site for pursuing development of isoform-selective inhibitors. We describe new lead compounds that will be optimized into the first selective and in vivoactive DGK-alpha inhibitor with potential applications for immuno-oncology. We also describe our efforts to use mass spectrometry-metabolomics to annotate substrate specificity of all 10 mammalian DGKs in live cells to identify biomarkers for future applications in studying T cell biology.
Recombinant human IL-15 (rhIL-15) in combination with anti-PD-L1 (Avelumab) in SIV infected rhesus macaques leads to the expansion of a subset of CXCR3+PD1−/low CD8 T cells
Authors: MOUSSA Maha , Cheng Jie , Chen Ping , Chen Hui , Li Tong , Qin Jing , Lifson Jeffrey D. , Sneller Michael C. , Krymskaya Ludmila , Godin Steven , Lane Clifford , Catalfamo Marta 
Published: January 2023

Abstract
The PD1/PD-L1 pathway contributes to the pathogenesis of HIV/SIV infection. Expression of PD1 by HIV/SIV-specific CD8 T cells can limit antiviral responses and expression of checkpoint receptors including PD1 by HIV infected cells can contribute to maintenance of viral reservoirs. In addition, reports have shown that IL-15 induces the expansion of effector memory CD8 T and NK cells. In this study, we hypothesized that expansion of memory CD8 T cells in the context of PD1/PD-L1 blockade may lead to enhanced anti-SIV responses. We design a study in which we evaluated the effect of rhIL-15 administered in cycles of ten days by continuous infusion in combination with weekly administration of anti-PD-L1 (Avelumab) over 24 weeks. After seven doses of anti-PD-L1 and completion of two cycles of rhIL15, cART therapy was interrupted and anti-PD-L1 administration continued to complete 24 weeks treatment. We found that administration of rhIL-15 in combination with anti-PD-L1 treatment was well tolerated. rhIL-15 infusions were associated with transient increases in proliferation of NK cells and effector memory CD8 T cells. rhIL-15/anti-PD-L1 treatment led to a sustained expression of CXCR3+ with low to undetectable expression of PD1on CD8 T cells, suggesting potential of increased tissue trafficking. Multidimensional single cell analysis showed that CXCR3+PD1−/low CD8 T cells secrete cytokine in response to viral antigens. Despite these clear biological effects, no impact was observed on rebound plasma viremia after cART discontinuation and during anti-PD-L1 administration. Futures studies should characterize the CXCR3+PD1−/low CD8 T cell subset expanded by this treatment to understand the potential impact of this subset in viral control.