bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bioengineering localized and controlled drug delivery to actuate Enhanced Costimulation Blockade and promote long-term transplant survival
Authors: Raimondi Giorgio , Majumder Poulami , Ivanova Vera , Calderon-Colon Xiomara , Mirdad Amr , Lozano Marcos Iglesias , Lee WP. Andrew , Brandacher Gerald , Schneider Joel , Patrone Julia 
Published: January 2023

Abstract
We recently demonstrated that a transient daily administration of the Jak-inhibitor Tofacitinib (Tofa) combined with CTLA4-Ig exerts what we call “enhanced costimulation blockade” and promotes long-term transplant survival in a mouse heart transplant model. To minimize the systemic exposure to Tofa and maximize its immune-modulatory effect, we explored two innovative drug delivery methods: a peptide-based hydrogel (Tofa-Hydro), and solid lipid nanoparticles (Tofa-SLN).
Tofa-Hydro is a safe injectable hydrogel. A one-time application of Tofa-Hydro in the area surrounding the transplanted heart (combined with systemic CTLA4-Ig) rendered long-term graft survival (MST=127d Vs ‘CTLA4-Ig only’ MST=36, P&lt;0.05). Importantly, application of Tofa-Hydro to a distal site ablated this synergistic improvement of survival, demonstrating the importance of the site of delivery. Tofa-SLN showed the unique property of accumulating in the lymphatics, reaching draining lymphoid tissues, and being taken up by immune cells. Tofa-SLN-conditioned dendritic cells were profoundly inhibited in their maturative response to LPS. These cells were less stimulatory for alloreactive T-cell and worked in concert with CTLA4-Ig to minimize T cell activation.
Our results highlight two promising strategies for controlled and localized immune modulation. Injectable hydrogels allow a sustained localized delivery that requires a fraction of the dose of Tofa required when delivered systemic. Tofa-SLN can be passively directed to lymphoid tissues and effectively target and modulate antigen presenting cells. Overall, these strategies can maximize the immunomodulatory impact of Tofa while minimizing its toxic side-effects.
Alloreactive T cells develop in the very early period following human stem cell transplantation and are characterised by a range of novel features
Authors: Moss P. , Verma Kriti , Zuo Jianmin , Malladi Ram 
Published: January 2023

Abstract
Allogeneic stem cell transplantation can cure some patients with hematopoietic malignancy but relies on development of a donor T cell alloreactive immune response which develops very early but is difficult to study due to intense lymphopenia. We characterised the phenotype and functional profile of T cells at day 14 in patients undergoing transplantation. T cells were present at very low number but displayed intense proliferation with strong cytokine production. Oligoclonal expansions at day 14 came to represent a substantial fraction of the established T cell pool and were recruited into tissues affected by graft-versus-host disease. Transcriptional analysis uncovered a range of potential targets for immune manipulation including OX40L, TWEAK and CD70. These findings reveal that recognition of alloantigen rapidly drives naïve T cells towards a unique phenotype characterised by intense functional activation. Moreover they demonstrate that early clonal T cell responses are recruited to sites of subsequent tissue damage and provide a range of novel targets for potential therapeutic immunomodulation.
Soy-enriched diet limits tumor progression and reduces myeloid derived suppressor cells in the murine TRAMP model of prostate cancer.
Authors: Mace Thomas A. , Loftus Shannon , King Samantha , Young Gregory , Geraghy Connor , Fareen Matthew R. , Ware Brandon , Olson Brian , Thomas-Ahner Jennifer , Clinton Steven K. , Lesinski Gregory B. 
Published: January 2023

Abstract
Epidemiological studies suggest diets enriched with soy confer a lower risk of prostate cancer. Data from our group and others indicate that soy has immunomodulatory properties which may contribute to their health promoting effects. Prior results from our group in a randomized phase II clinical trial revealed that men with asymptomatic prostate cancer on a soy-enriched diet had significantly reduced systemic inflammatory cytokines and myeloid-derived suppressor cells (MDSCs). We hypothesized that a soy-enriched diet limits systemic immunosuppression by reducing MDSC expansion. To understand how dietary soy modulates immune biomarkers in prostate cancer, we administered TRAMP (transgenic adenocarcinoma of the mouse prostate) mice a soy-enriched (n=28) or control diet (n=32) for 16 weeks. TRAMP mice administered a soy-enriched diet had a reduced tumor burden compared to control fed mice (p=0.0419). Splenic CD11b+Ly6GloLy6C− granulocytic (p=0.0195), CD11b+Ly6C+Ly6G− monocytic (p=0.0005), and total MDSCs (p=0.0009) were significantly reduced in TRAMP mice on a soy-enriched diet compared to control diet. However, we did not observe differences in circulating CD4+CD25+Foxp3+ T-regulatory cells or other circulating immune subsets such as CD4+ T cells, phenotypically defined T helper-1 (CD4+CXCR3+CCR4−CCR6−)/T helper-2 (CD4+CCR4+CXCR3+CCR6−) T cells, CD8+ T cells, and NK cells. Ongoing studies are examining plasma cytokine levels and immunosuppressive cell populations in the tumor microenvironment of mice from this study. Together, these data indicate that soy as a dietary natural product may have a modulatory effect during chemopreventive or immunotherapy in the setting of prostate cancer.
Opposing roles of CD2 and 2B4 in iNKT cell cytotoxic responses
Authors: Das Rupali , Lee Hyun Hee , Gohl Trevor , Mack Ryan , Griffin Ryan 
Published: January 2023

Abstract
Invariant natural killer T cells (iNKTs) comprise a unique lineage of innate-type lipid-reactive T lymphocytes with important roles in host immunity, including defense against specific pathogens and cancers. iNKTs rapidly produce cytokines and mount potent cytotoxic responses following T cell receptor (TCR) engagement, but the mechanisms that control iNKT cell functions remain poorly understood. CD2 and 2B4 are receptors on iNKTs that both bind to CD48 on target cells and modulate immune activation by facilitating cell-cell interactions and transducing intracellular signals. To determine their roles during iNKT cell activation, we examined the TCR-induced killing activity of murine iNKTs that lacked expression of CD2 or 2B4. Through these studies, we observe that Cd2−/− iNKTs fail to kill target cells while 2b4−/− iNKTs exhibit significantly enhanced cytolysis, both in vitro as well as in vivo. These novel findings suggest that CD2 is a positive and 2B4 a negative regulator of TCR-induced iNKT cell functions. We are currently investigating whether 2B4 exerts its inhibitory role by directly competing with CD2 for CD48 binding and/or by inducing a negative signal. As CD1d (the ligand for the invariant TCR) and CD48 (the ligand for CD2 and 2B4) are both widely expressed on hematopoietic tumors, further elucidation of the mechanisms by which CD2 and 2B4 regulate TCR-induced iNKT cell anti-tumor activity is of significant scientific and clinical importance. The results obtained from these studies will have an impact by establishing new paradigms for iNKT signaling and offering insights into how iNKTs can be best activated to enhance host immunity and treat hematopoietic cancers such as leukemia and lymphoma.
Engineering Chimeric Antigen Receptor T-cells Against Immune Checkpoint Inhibitor PD1 and PD-L1 for Treating Pancreatic Cancer
Authors: Fan Ming huei , Fan Kai Fang , Liu Chao-Lien 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDA) is a serious and aggressive disease with 5-year survival rate of 6%, the lowest of any tumor type routinely checked. Immune checkpoint inhibitor pathway PD1 (Programmed-Death-1) /PD-L1 (Programmed-Death-1 Ligand-1) axis is responsible for inhibitory T cell signaling and contributes to the maintenance of immune homeostasis. PD1/PD-L1 pathway plays an essential role in the tumor microenvironment and blockade of PD1/PD-L1 by monoclonal antibodies has success in the treatment of several tumor types. However, even though PD-L1 is expressed on pancreatic cancer cells, current clinical trial using PD1/PD-L1 antibodies blockade failed to show therapeutic effects. In this study, we first generated both PD1ACR (activating chimeric receptor)-T and PDL1CAR (chimeric antigen receptor)-T cells, elucidate the molecule mechanisms leading to the development of effector cells as well as to test the cytotoxicity function of these CAR-T cells. CFPAC1, a high PDL1-expressed cell line, was checked among four pancreatic cancer cell lines including, CFPAC1, SU8686, Capan1 and ASPC1 using flow cytometry and western blot analysis. The in vitro cytotoxicity activities towards CFPAC1 were significantly in the PD1ACR-T (35%) and PDL1CAR-T (55%) at E/T (effector/tumor) ratio of 10 compared to the controls. Furthermore, in tumor xenografts of NSG mice, both PD1ACR-T and PDL1CAR-T treated mice groups showed significantly suppressed tumor growth at 59% and 77%, respectively. Therefore, we concluded that PD1/PDL1 mediated CARs possess dramatic cytotoxic activities towards PDL1-expressed pancreatic cancers both in vitro and in vivo, which strongly support the potential clinical immunotherapeutic applications.
Regulatory T cells modulate immune cells and promote tumor growth in prostatic tumor microenvironment
Authors: Kumar Sanjay , Stokes James , Malik Shalie , Singh Udai P. , Singh Rajesh , Ponnazhagan Selvarangan , Manne Upender , Singh Santosh Kumar 
Published: January 2023

Abstract
Several tumors express antigens that can be recognized by T lymphocytes, suggesting that T cell-mediated immune response may play a critical role in anti-tumor immunity. Studies demonstrated that Regulatory T cells (TR) suppress the functions of immune cells, which are capable to recognized and eliminate tumor cells. However, there exists an utmost demand for further research how immune cells (CD8+ cells, NK cells, etc.) fail to suppress tumor growth and proliferation in the prostatic tumor microenvironment. To investigate this, tumor induction studies in C57/BL-6 mice using transgenic adenocarcinoma of mouse prostate (TRAMP cell lines: -C1, -C2 and -C3) as tumor models were performed. TRAMP-C3 did not form tumor while TRAMP-C1 and TRAMP-C2 cells are tumorigenic. For tumor induction, mice were administered with TRAMP (C1-3) cells. Mice were sacrificed by cervical dislocation when tumor sizes reached ~18–20mm in diameter. A single cell suspension (obtained from spleen, draining lymph node, and tumor) was stained with various cell surface/intracellular immune markers/cytokines to enumerate CD8+ T/TR/NK/Ly6G/Ly6C+ -MDSC cells. Additionally, the expression of TGF-β, IFN-γ, IL-4, IL-17, PD-1 and PDL-1 and cell cycle analysis for CD8+ T cells was also carried out. Our findings suggest that recruitment of suppressive TR and Ly6G/Ly6C+ -MDSC cells and high expression TGF-β and PD-1 and lower expression of NKG2D and IFN-γ and low number of NK and CD8+ cytotoxic T cells in spleen, draining LN and tumor of TRAMP-C1 and TRAMP-C2 (as compared to TRAMP-C3 mice) facilitated tumor growth. Therefore, the expression of PD-1 and TGF-β on TR suggest that TR might be playing a significant role in facilitating tumor growth and progression.
Uncoupling Therapeutic from Immunotherapy-related Adverse Effects for Safer and Effective Anti-CTLA-4 Antibodies in CTLA4 Humanized Mice
Authors: DU XUEXIANG , Tang Fei , Liu Mingyue , Liu Yang , Zheng Pan 
Published: January 2023

Abstract
Anti-CTLA-4 monoclonal antibodies (mAbs) confer cancer immunotherapeutic effect (CITE) but cause severe immunotherapy-related adverse effect (irAE). The irAE greatly limited application of immunotherapy targeting the CTLA-4 molecules. However, targeting CTLA-4 has shown remarkable long-term benefit and thus remains a valuable tool for cancer immunotherapy, especially if the irAE can be brought under control. A major obstacle to this goal is lack of animal model that recapitulate clinical irAE. Here we report that mice with the humanized CTLA4 gene recapitulated cardinal features of clinically observed irAE and CITE in humans. Of two anti-human CTLA-4 mAbs with comparable CITE, Ipilimumab but not L3D10 induced severe irAE, when used either alone or in combination with anti-PD-1. The irAE, but not CITE, corresponds to systemic T cell activation and reduced Treg/Teff ratios among autoreactive T cells. While Ipilimumab+anti-PD-1 induces irAE and CITE in mice homozygous for human CTLA4, the same treatments induced CITE without irAE in mice heterozygous for human and mouse CTLA4 alleles, thus revealing a distinct genetic requirement for irAE and CITE. Humanization of L3D10 that leads to loss of blocking activity further increased safety without affecting the therapeutic effect. Taken together, our data demonstrate that complete CTLA-4 occupation, systemic T cell activation and preferential expansion of self-reactive T cells are dispensable for tumor rejection but correlate with irAE, while blocking B7-CTLA-4 interaction impacts neither safety nor efficacy of anti-CTLA-4 antibodies. These data provides important insights for clinical development of CTLA-4 targeting immunotherapy.
Identification and characterization of novel mediators of tumor-induced T-cell dysfunction
Authors: Markowitz Geoffrey J. , Philip Mary , Wong Stephen , Schietinger Andrea , Mittal Vivek 
Published: January 2023

Abstract
Immunotherapies targeting T-cell functionality, such as anti-PD-1, have been shown to have efficacy in numerous cancers, including non-small cell lung cancer (NSCLC). However, treatment with anti-PD-1 induces responses in only a subset of patients. This raises the possibility that other effectors of T-cell dysfunction remain to be elucidated. To evaluate possible novel mediators of T-cell dysfunction, we isolated CD8+ T-cells from orthotopic KrasG12D/p53−/− murine lung adenocarcinomas (HKP-1) that exhibited either stable or progressive disease as a function of tumor growth, both in the presence and absence of anti-PD-1 therapy. RNAseq analysis revealed increased expression of numerous genes including Tim3, PD-1, Lag3, 2B4, and CISH, and decreased expression of GzmB and Eomes in CD8+ T-cells isolated from progressive tumors indicative of a dysfunctional state, as well as multiple novel genes not previously associated with T-cell dysfunction. To explore the functional role of the novel candidate genes, we have developed in vitro and in vivo assays evaluating cytotoxicity, cytokine production, proliferation, and tumor growth, leveraging Ova expression in our HKP-1 cell line (KP1-Ova). We have begun to target this unique set of candidate genes associated with dysfunction in tumor-specific T-cells through RNAi/shRNA and adoptive transfer approaches to facilitate therapeutic reprogramming and enhanced T-cell-mediated immunity in lung cancer. We expect to provide novel strategies for designing rational immunotherapies targeting dysfunctional tumor-specific CD8+ T-cells to generate durable anti-tumor immune responses for the treatment of lung adenocarcinoma.
Manipulating the epigenetic framework of T cells with histone deacetylase inhibitors for more robust and durable anti-tumor responses
Authors: McCaw Tyler R. , Liu Mingyong , Li Mei , Starenki Dmytro , Cooper Sara , Arend Rebecca , Forero Andres , Buchsbaum Donald , Randall Troy 
Published: January 2023

Abstract
Malignant cells often harbor an imbalance in histone acetyltransferase and histone deacetylase (HDAC) activity, which can be disrupted by HDAC inhibitors. We hypothesized that HDAC inhibition can boost anti-tumor T cell responses by modifying tumor cell phenotypes as well as directly altering the transcriptional framework of tumor-specific T cells. To test this, we treated two murine breast cancer cell lines, TS/A and 4T1, with class I (entinostat), or pan-specific (panobinostat), HDAC inhibitors. Culture of tumor cells with either inhibitor increased surface expression of MHC and costimulatory molecules as well as T cell chemoattractants. Tumor growth was significantly impaired by HDAC inhibition in a manner absolutely dependent on adaptive immunity, specifically CD8 T cells and IFNγ production, but not CD4 T cells or B cells. Indeed, CD8 T cell production of effector cytokines was dramatically augmented. This enhanced effector function was paralleled by an increase in T-bet and a decrease in Eomes, consistent with a transcriptional program that is less susceptible to exhaustion. We also found that simply adjusting timing of HDAC inhibition relative to T cell activation could abolish anti-tumor effects or lead to rejection in nearly 50% of mice. These findings corresponded with a significant shift in the responding T cell repertoire. Collectively, our data suggests that HDAC inhibition reprograms the transcriptional profile of T cells in ways that impact priming, improve effector functions, and reduce susceptibility to exhaustion. Thus, appropriately timed HDAC inhibition may synergistically potentiate existing tumor immunotherapies, like T cell reinvigoration strategies.
Immunosuppressive effects of sMIC abrogate immunotherapy efficacy in a mouse model of breast cancer
Authors: Berkley Amy M. , Toy Evangeline , Cook Ryan , Ye Zhengmao , Grogan Jane , Schartner Jill , Kim Jeong 
Published: January 2023

Abstract
Expression of the NKG2D activating receptor on CD8+ T cells and NK cells is thought to play an important role in detection and clearance of tumor cells. The soluble version of an NKG2D ligand in humans, MHC I chain-related molecule (sMIC), has been correlated with poor prognosis in multiple human cancers and hypothesized to serve as a tumor immune evasion strategy. Yet, it is unclear whether sMIC impairs the activity of T cell directed immunotherapies including checkpoint inhibitors. We utilized a syngeneic mouse model of breast cancer, EMT6, as well as EMT6 engineered to express high levels of sMIC to examine the impact of sMIC on responses to anti-PDL1/anti-TIGIT combination immunotherapy. Following treatment with anti-PDL1/anti-TIGIT combination immunotherapy, WT EMT6 tumors were rejected and tumor associated T cells and NK cells isolated from these tumors showed increased IFNγ secretion. In contrast, mice bearing EMT6 tumors expressing sMIC and treated with the same regimen were unable to reject tumors. Consistent with this observation, CD8+ and NK cell TILs from EMT6. sMIC tumors showed decreased ability to produce IFNγ compared to those from WT EMT6 tumors. Taken together these results suggest the importance of targeting the MIC-NKG2D pathway to further improve combination immunotherapy efficacy in patients.
Canonical Wnt signaling in Treg contributes to colon cancer
Authors: Osman Abu , Saadalla Abdulrahman , Quand Jasmin , Gounari Fotini , Pavelko Kevin , Khazaie Khashayarsha 
Published: January 2023

Abstract
Regulatory T-cells (Tregs) play a dual role in colon cancer, promoting immune tolerance and suppressing inflammation. Two types of Tregs infiltrate the gut. Thymic derived Helios+CD4+Foxp3+ Tregs have high affinity for self-antigens, including tumor-antigens and protect the tumor, while Helios−CD4+Foxp3+ extra-thymic Tregs that are generated in the gut have microbial specificity and suppress harmful inflammation. In colon cancer, there is a preferential expansion of a population of Helios−CD4+Foxp3+RORγt+ Tregs with enhanced T-cell suppressive properties and pro-inflammatory properties that promote tumor growth. We showed that this population of Tregs in cancer patients expresses elevated levels of β-catenin. To test the role of canonical Wnt signaling in the generation of pro-inflammatory Tregs, we stabilized β-catenin or deleted its DNA binding partner TCF-1 in Tregs. In both instances expression of RORγt was upregulated. While the stabilization of β-catenin led to loss of Treg functions, ablation of TCF-1 reproduced the phenotype and functional characteristics of Tregs in colon cancer. The TCF-1 deficient Tregs were pro-inflammatory, potent T-cell suppressive, and tumor promoting.
IL-21 selectively protects CD62L+ NKTs from activation-induced cell death during ex vivo expansion and enhances antitumor activity of NKT cell therapy in vivo
Authors: Ngai Ho , Tian Gengwen , Courtney Amy N. , Marinova Ekaterina , Huang Wei , Guo Linjie , Liu Bin , Metelitsa Leonid S. 
Published: January 2023

Abstract
Vα24-invariant natural killer T cells (NKTs) have potent antitumor properties that can be exploited for use in cancer immunotherapy. This requires rigorous protocols for ex vivo expansion of primary NKTs that preserve their longevity and function. We recently reported that CD62L+ NKTs persist longer and have better antitumor activity in vivo than CD62L− counterparts. However, the conditions governing preservation of CD62L expression in NKTs during expansion remain largely unknown. Comparative gene expression analysis of CD62L+ and CD62L− NKTs revealed significantly higher IL-21R expression in the former subset, which was confirmed at the protein level by flow cytometry. Hence, we hypothesized that IL-21 preferentially supports CD62L+ NKTs. We expanded primary human peripheral blood NKTs by stimulating in vitro with cognate antigen, α-galactosylceramide, and supplemented the culture with IL-2, IL-21, or both cytokines. In contrast to IL-2, IL-21 alone failed to support NKT cell expansion, but combined treatment with IL-2 and IL-21 produced more NKTs and preserved a significantly higher frequency of CD62L+ cells. We further demonstrated that IL-21 selectively downregulates BIM in CD62L+ NKTs and protects the subset from activation-induced cell death. Functionally, IL-2/IL-21-expanded NKTs secreted more TH1 cytokines and were more cytotoxic against lymphoma cells, which was associated with granzyme B upregulation by IL-21. Following adoptive transfer to NSG mice, IL-2/IL-21-expanded NKTs persisted significantly longer and had higher therapeutic efficacy in a lymphoma model compared with IL-2-expanded NKTs. Our results instruct inclusion of IL-21 in NKT-cell expansion protocols for cancer immunotherapy applications.
Drugging drug resistance with bystander-assisted immunotherapy.
Authors: Mancini Rock , Nielsen Amy E. , Hantho Joseph D. , Burt Anthony J. 
Published: January 2023

Abstract
Acquired drug resistance is a longstanding challenge that reduces the efficacy of chemotherapeutic drugs. In contrast, the efficacies of modern immunotherapies do not directly correlate to the prevalence of chemo-resistant phenotypes. Here we improve upon this paradigm by targeting the action of immunotherapeutics to two general mechanisms of acquired drug resistance: irregular metabolism and drug efflux mediated by the ABC superfamily of transport proteins. In chemo-resistant prostate cancer cells, we demonstrate that these two mechanisms act in concert to selectively metabolize our newly developed class of enzyme-directed prodrug to the Toll-Like Receptor 7 immunotherapeutic Imiquimod. Following metabolism, liberated Imiquimod undergoes drug efflux to the extracellular space where it activates bystander immune cells in local proximity. In-vitro, we characterize this process of Bystander-Assisted ImmunoTherapy (BAIT) using an AT3B-1 chemo-resistant prostate cancer model with RAW-Blue and JAWSII reporter immune cell lines. Co-culture of AT3B-1 cancer cells with reporter immune cells results in immunogenicity mediated selectively by cancer cells. This results in enhanced NF-κB transcription, expression of cell surface markers, and secreted cytokines indicative of a cell-mediated immune response. Our prodrug is non-immunogenic with healthy cells alone and the enzyme-directing groups are stable for over 3 days in serum. Taken together, our results demonstrate that BAIT co-opts common mechanisms of drug resistance to elicit immunogenicity mediated by cancer cells themselves. We anticipate that BAIT will find use as a new mechanism of action that exploits drug resistant phenotypes to generate an immune response.