bluuebunny/crossref_metadata_2025_split huggingface dataset:
Stem cell-derived tissue-associated regulatory T cells suppress autoimmune diabetes
Authors: Haque Mohammad Shamsul 
Published: January 2023

Abstract
CD4+ regulatory T cells (Tregs) are essential for normal immune surveillance, and their dysfunction can lead the development of autoimmune diseases, such as type-1 diabetes. Pluripotent stem cells (PSCs) can be utilized to obtain a renewable source of healthy Tregs to treat autoimmune diabetes as they have the ability to produce almost all cell types in the body, including Tregs. However, the right conditions for the development of antigen (Ag)-specific Tregs from PSCs (i.e., PSC-Tregs) have not been fully defined. In this study, we developed a new approach to generate functional Ag-specific Tregs from induced PSC (iPSCs), i.e., iPSC-Tregs, which have the ability to suppress autoimmunity in a murine model of autoimmune diabetes in Ag-associated fashion. We retrovirally transduced murine iPSCs with a DsRed reporter construct containing genes of major histocompatibility complex (MHC) II (I-Ab)-restricted ovalbumin (OVA)-specific T cell receptor (TCR) and the transcriptional factor FoxP3. We in vitro differentiated the DsRed+ iPSC into OVA-specific iPSC-Tregs with an OP9 stromal cell line expressing Notch ligands DL1, DL4 and I-Ab in the presence of recombinant cytokines of rIL-7 and rFlt3L. We visualized the expression of CD3, TCR, CD4, CD25, and CTLA-4 on OVA-specific iPSC-Tregs. Moreover, adoptive transfer of such Tregs dramatically suppressed autoimmunity in a well-established OVA-induced model of autoimmune diabetes (RIP-OVA × OT-I double transgenic mice), including the inflammation and prevents the insulin secreting pancreatic beta cells from destruction. Our results indicate that PSCs can be used to develop Ag-specific Tregs, which have a therapeutic potential for autoimmune diabetes.
PD-L1/PD-1 Immunotherapy Modulates Effector T Cells Homeostasis and Function in Murine Pancreatic Cancer
Authors: Rivera Ivannie Ortiz , Iclozan Cristina , McGill Andrew , Ghansah Tomar 
Published: January 2023

Abstract
Pancreatic Cancer (PC) is one of the most aggressive and deadliest types of cancer, and it is projected to be the second leading cause of cancer-related deaths in the U.S. by the year 2030.
PC evades immune surveillance by disrupting the immune homeostasis of effector T cells (Teff). T cells homeostasis is critical for proper anti-tumor immune responses. Preliminary flow cytometry data revealed that murine pancreatic (Panc02) cancer cells produce inflammatory soluble factors and in addition, express Major Histocompatibility Complex class I (MHC-I) and Programmed Death Ligand 1 (PD-L1) and 2 (PD-L2). PD-L1/2 can bind to PD-1 receptors on Teff and induce apoptosis or anergy, dampening the anti-tumor immunity. Cytokine bead array and flow cytometry analysis of serum from peripheral blood from C57BL/6 mice injected with Panc02 cells (TB mice) showed a significant increase in inflammatory factors compared to Control (CTRL) mice. Additionally, Teff from splenocytes from TB mice showed significant reduction in Teff percentages compared to CTRL mice, in vitro. In this study, CD3+ cells from CTRL mice were co-incubated with Panc02 cells pre-treated with anti-PD-L1 antibody (ab) to evaluate the modulation of Teff and cytokines production, in vitro. Mice were inoculated with the Panc02 pre-treated cells with anti-PD-L1 ab, to evaluate the modulation of Teff and cytokines production, in vivo. The results from this project may lead to the identification of new immunotherapeutic strategies to stabilize Teff homeostasis and function that could increase anti-tumor immune responses and combat PC progression.
Control of tumor growth by TSLP during colorectal cancer
Authors: de Jesus-Carrion Steven , Ziegler Steven F 
Published: January 2023

Abstract
Thymic stromal lymphopoietin (TSLP) is a cytokine involved in promoting tumor growth during breast and pancreatic cancer by promoting Th2 cell-mediated inflammation. Despite being essential to maintain Treg cell homeostasis in the mouse and human intestine, a role for TSLP in colorectal cancer has never been shown. To determine if TSLP affects tumor growth during colorectal cancer, we utilized a murine model of colitis-associated colorectal cancer. TSLP deficient mice exhibited decreased tumor numbers when compared to the WT littermate controls. Moreover, we found that tumors in TSLP−/− mice were also significantly smaller in size than WT mouse tumors. TSLP was mainly localized in the tumor tissue, indicating that it might be produced by cancer or cancer-associated cells. Further, TSLP receptor (TSLPR) expression was significantly upregulated after development of cancer. To determine if TSLP signals directly on the tumor cells, we utilized Villincre TSLPRflox mice in which the TSLPR expression is absent only in intestinal epithelial cells (IECs). Consistent with the TSLP−/− mice, lack of TSLP signaling on IECs led to a decrease in tumor number as well as tumor size, suggesting that TSLP signals directly on the IECs to promote tumor growth. Overall, these data show a novel role for TSLP in controlling tumor progression during colorectal cancer and identify it as a potential target for immunotherapy intervention.
Interleukin-10 Attenuate Tumor Growth by Inhibiting Interleukin-6/Signal transducer and activator of transcription 3 Axis on Myeloid-Derived Suppressor Cells
Authors: Ko Hyun-Jeong , Kim Seong-Ryeol , Song Jae-Hyoung , Lee Bo-Ra 
Published: January 2023

Abstract
Interleukin-10 (IL-10) is a well-characterized anti-inflammatory cytokine, but its role in anti-cancer immunity is still controversial. When IL-10 knockout (IL-10−/−) mice were subcutaneously injected with TC-1 cancer cells expressing HPV E6 and E7 proteins, tumor growth was accelerated in IL-10−/− mice than wild type (WT) mice. Interestingly, the production of interleukin-6 (IL-6) was significantly increased in IL-10−/− mice than in WT mice. The blockade of IL-6 by systemic administration of anti-IL-6 receptor (IL-6R) monoclonal antibody (mAb) inhibited the tumor growth, and inhibited the generation of myeloid-derived suppressor cells (MDSCs) which were increased in IL-10 deficiency. MDSCs and tumor cells from IL-10−/− mice had increased phosphorylated-signal transducer and activator of transcription 3 (p-STAT3) level and the treatment of mice with STAT3 inhibitor, S3I reduced the tumor growth and inhibited the expansion of MDSCs. Accordingly, S3I also reduced tumor necrosis factor-a (TNF-a) and IL-6 in tumor. Although S3I treatment relieved T cells from suppression by reducing the number of MDSCs, it seems that the proportion of MDSCs having M2 macrophage phenotype was increased among myeloid cells since the transcription of Arg1, Mgl1, Mgl2, and, Fizz1, were significantly increased and the expression of some suppressive functional mediators of MDSCs, including Arg1 and iNOS was up-regulated. Combined treatment of anti-IL-6 mAb and S3I could further suppress the generation of M2-type MDSC and regulate immunosuppressive function of MDSCs, and finally inhibited tumor growth more efficaciously. These results suggested that combined inhibition of IL-6 and STAT3 could be a potential candidate as an anti-cancer strategy.
Hypoxic tumor microenvironment of head and neck cancers downregulates Kv1.3 channels in tumor infiltrating lymphocytes
Authors: Chimote Ameet A , Hajdu Peter , Sfyris Alexandros M , Casper Keith A , Conforti Laura 
Published: January 2023

Abstract
The clinical outcome of solid malignancies depends on the ability of cytotoxic CD8+ T cells to infiltrate the tumor and destroy cancer cells. Unfortunately, tumor infiltrating lymphocytes (TILs) exhibit attenuated functions. Kv1.3 channels are vital to the T cell effector functions as they regulate the Ca2+ signaling necessary for cytokine and granzyme production. We have previously reported that the expression of functional Kv1.3 channels in healthy T cells is suppressed by hypoxia. Solid tumors like head and neck squamous cell carcinomas (HNSCC) are characterized by a hypoxic tumor microenvironment (TME). Herein we investigated whether the hypoxic TME in HNSCC inhibits Kv1.3 expression and function in TILs. Electrophysiological experiments showed a 70% reduction in functional Kv1.3 channels in freshly isolated HNSCC TILs compared to peripheral blood T cells. Kv1.3 currents in TILs ranged from 0 to 600 pA with a median of 90 pA. Immunofluorescence analysis of Kv1.3 and HIF1α (marker of hypoxia) in HNSCC tumor sections revealed that 88% of the CD8+ TILs expressing high Kv1.3 (Kv1.3high: above the median Kv1.3 fluorescence in all TILs) have a low HIF1α level. Furthermore, CD8+ cells preferentially localize in the stroma (68% of the TILs) than in the tumor epithelia (32%), and only 35% of the CD8+ cells in the tumor epithelia are Kv1.3high compared to 55% in the stroma, with a 20% reduction in the median Kv1.3 fluorescence of CD8+ cells in the tumor epithelia compared to the stroma. Overall, our data indicate that exposure of lymphocytes to the hypoxic HNSCC TME leads to decreased Kv1.3 expression. This reduction in Kv1.3 may contribute to the attenuated cytotoxicity of TILs and improper immune surveillance in solid tumors. (NIH2R01CA095286)
The surface alpha-enolase targeting as a novel immunotherapeutical strategy for pancreatic cancer
Authors: Cappello Paola , Principe Moitza , Chattaragada Michelle Samuel , Curto Roberta , Giordano Daniele , Roux Cecilia , Novelli Francesco 
Published: January 2023

Abstract
Pancreatic Ductal Adenocarcinoma (PDA) is a very aggressive tumor for which effective therapeutical strategies are still lacking. The global five-years survival is of 5% and surgery is the only potentially curative treatment. The PDA-associated antigen α-enolase (ENO1), beside its glycolytic function, acts as a plasminogen receptor, promoting activation into plasmin, involved in extracellular matrix degradation. Antibodies against ENO1 are detected in more than 60% of PDA patients and correlate with a better prognosis. Furthermore, anti-ENO1 antibodies are induced in mice after ENO1-DNA vaccination. We observed an increased ENO1 expression on the cell surface of both PDA and myeloid cells (MDSC), suggesting a role in tumor progression and spreading. The immunotherapy represents a chance to selectively target PDA cells and MDSC. Mouse anti-human ENO1 monoclonal antibody (mAb) inhibits plasminogen-dependent invasion of human PDA cells and metastatic spreading in immunosuppressed mice, as well as MDSC adhesion to pancreatic endothelial cells and in vitro and in vivo migration. Similarly MDSC arginase activity and secretion of IL-6 decrease after ENO1 triggering, while anti-ENO1 treatment does not affect costimulatory molecule expression and MDSC suppression functions. Notably, a single administration of Adeno-Associated Virus (AAV)-expressing an anti-ENO1 mAb reduced the number of lung metastases in immunosuppressed mice injected with PDA cells. In conclusion, the antibodies anti-ENO1 may inhibit PDA cell and myeloid cell invasion and modulate T cell response, making the immunotherapy more effective.
Quantitation of sub-visible particles in therapeutic protein formulations using Imaging Flow Cytometry
Authors: Probst Christine Elizabeth , Zeng Yuanchun , Zhu Rong-Rong 
Published: January 2023

Abstract
Monitoring particles within therapeutic protein formulations is necessary to evaluate drug safety and efficacy. Current regulations require analysis of particles greater than 10 μm, however quantitation of particles 1–10 μm is increasingly expected due to potential immunogenicity. These small particles are difficult to measure with current methods, and new methods must be developed. Amnis Imaging Flow Cytometry (IFC) is an existing technique that is conceptually well-suited for this application because provides: (1) Sensitive detection particles across a wide size range (1–100 μm), (2) Direct particle size/shape measurements from brightfield imagery (3) Particle identification using fluorescent dyes, (4) High sampling efficiency (95%) (5) Low sample volume requirement (20 μL). Here we demonstrate IFC may be used to measure particles common to this application, including protein aggregates, silicone oil droplets, polystyrene bead standards, and newly developed protein surrogate standards. IFC provided sensitive detection of particles as small as 100 nm and could be used to discriminate protein aggregates from silicone oil. Interestingly, IFC analysis of one sample revealed transparent protein aggregates invisible using current gold-standard methods. The results demonstrate that IFC is a useful orthogonal technique to current methods for interrogating small and translucent particles within protein formulations.
ImmTACs: Bi-specific TCR-based reagents for targeted cancer immunotherapy.
Authors: Jakobsen Bent 
Published: January 2023

Abstract
Immune system based therapies constitute a promising class of treatment for many types of cancers. Whilst T cells can mediate tumor destruction, their effectiveness is limited due to negative thymic selection, down-regulation of HLA expression by cancer cells and an immunosuppressive microenvironment. To overcome the poor immunogenicity of tumors, Immunocore has developed ImmTACs (immune mobilizing monoclonal T cell receptors (mTCR) against cancer). A soluble mTCR, engineered to recognise a given tumor associated peptide-HLA complex with exceptionally high sensitivity and specificity, redirects host polyclonal T cells via an anti-CD3 antibody fragment, facilitating targeted T cell recognition of tumors. Antigens that exist on tumor cells but not on normal cells are rare, thus the selection and validation of appropriate target antigens and the testing of ImmTACs for specificity is critical. Antigen candidates are selected based on their levels of expression in cancer vs healthy tissues by RT-PCR and the presence of the targeted peptide on the cell surface is confirmed by mass spectrometry. As ImmTACs are specific for humans, efficacy, specificity and off target effects are determined through a detailed molecular and cellular testing programme, using antigen positive tumor cells and HLA appropriate primary human cell lines representing a range of tissues. IMCgp100, the lead ImmTAC, is currently in a Phase I/II clinical trial for the treatment of advanced melanoma. The maximum tolerated dose has been established and emerging data demonstrate several durable responses. IMCgp100 is well tolerated and there is evidence of T cell mobilisation in the tumor microenvironment, release of cytokines and tumor shrinkage.
Galectin-3 inhibition using novel inhibitor GR-MD-02 improves survival and immune function while reducing tumor vasculature
Authors: Linch Stefanie N , Kasiewicz Melissa J , McNamara Michael J , Hilgart-Martiszus Ian F , Farhad Mohammad , Traber Peter G , Redmond William L 
Published: January 2023

Abstract
Immunosuppression and reduced cytotoxic function of tumor-infiltrating lymphocytes (TIL) are major obstacles to creating effective therapies for patients. It is known that Galectin-3 (Gal3), a lectin family member, is expressed and secreted by numerous cancers and immune cell subsets. Serum Gal3 expression is higher in patients with metastatic versus non-metastatic disease, and is associated with reduced survival in metastatic melanoma. Furthermore, Gal3 has been implicated in disease progression via the promotion of angiogenesis and metastasis, the inhibition of TIL function, and promoting M2 polarization of macrophages and mobilization of myeloid cells from the bone marrow to promote a metastatic niche within the tumor.
We hypothesized that Gal3 inhibition would improve TIL function while inhibiting the growth and metastasis of tumors. Through collaboration with Galectin Therapeutics, we tested Gal3 inhibition using the novel Gal3 inhibitor, GR-MD-02, with agonist anti-OX40 therapy. We observed that Gal3 inhibition/OX40 agonism improved survival in the MCA-205 sarcoma and 4T-1 mammary carcinoma models, and prolonged survival in the TRAMP-C1 prostate cancer model. Importantly, Gal3 inhibition/OX40 agonism reduced lung metastases in the 4T-1 model. Within the tumor, we observed an increase in the number of proliferating and IFNg-producing TIL. Gal3 inhibition/OX40 agonism was also associated with a decrease in tumor vasculature, as determined by CD31 staining by IHC within the tumors. These studies, along with clinical data from a liver fibrosis trial, supported testing GR-MD-02 in combination with additional immunotherapies to augment T cell function in patients with metastatic melanoma.
Chlamydia trachomatis Recombinant MOMP Encapsulated within PLGA 85/15 Nanoparticles Activates Mouse Dendritic Cells by Triggering Enhanced Co-stimulatory Molecules and T Helper 1 Type Cytokines
Authors: Sahu Rajnish , Dixit Saurabh , Duncan Skyla , Singh Shree R , Dennis Vida 
Published: January 2023

Abstract
Chlamydia trachomatis (CT), the leading cause of bacterial sexually transmitted diseases, is of public health significance because it causes considerable morbidity and socioeconomic burden worldwide. The quest for vaccine development against this pathogen has escalated to reduce its occurrence and global economic burden. Recently nanovaccines have emerged as attractive therapeutics due to flexibility in their formulation, biocompatibility and adjuvant properties. We have developed a nanovaccine against CT employing its MOMP (major outer membrane protein) and encapsulating it in PLGA [poly (D, L-lactic-co-glycolic acid) 85/15 nanoparticles. We hypothesized that encapsulated MOMP will be effective in activating dendritic cells (DCs) for induction of Th1 immune responses, which are needed for protection against CT. Exposure of mouse DCs to encapsulated MOMP at concentrations of 0.01 to 1 μg/mL revealed marked IL-6 and IL-12p40 secretions, with the latter being 20-fold more abundant. Encapsulated MOMP induced lower IL-10 and TNF levels with an elevated IL-12p40/IL-10 ratio, suggesting a predominant Th1 response. Time-kinetics studies (4 to 96 hr) showed increasing IL-12p40 concomitant with decreasing IL-10 concentrations. Analyses of CD11c+ DCs by FACS indicated heightened expression of the co-stimulatory molecules, CD80, CD86 and CD40 in response to encapsulated MOMP stimulation. TaqMan qPCR for mRNA transcripts validated the upregulation of TLR-2, CD80, CD86 and particularly CD40, which are important for adaptive immune responses. Our data suggest that encapsulated MOMP is a potential nanovaccine candidate by triggering enhanced co-stimulatory molecules and Th1 responses, which are required to protect against CT.
Enhanced protective efficacy of a PRRSV DNA vaccine by absorption onto biodegradable PLGA/BPEI nanoparticle
Authors: Li Bin , Du Luping , He Kongwang 
Published: January 2023

Abstract
Porcine reproductive and respiratory syndrome (PRRS) is one of economically devastating viral diseases of pigs, which has recently caused catastrophic losses in swine industry worldwide. Developing preventive measures to control PRRS outbreaks has been a challenge to the swine industry worldwide. Current PRRSV vaccination strategies provide only a limited protection. The safer and more effective new vaccines are urgently needed.
Nanoparticles (NPs) as vaccine carriers have a broad application prospect in design of vaccine systems. PLGA nanoparticle-mediated delivery of vaccines has shown to be effective in eliciting protective immune response, administered by either mucosal or systemic routes. PEI, due to its high positive charge density, has been widely used as a cationic polymer for the preparation of the cationic PLGA particles.
In this study, a synthetic ORF5 gene (SynORF5) of PRRSV as a model vaccine (pcDNA3.1-SynORF5), which has been employed and demonstrated that it could significantly induce the humoral and cellular immunity responses, was absorbed onto the PLGA/PEI nanoparticles. Moreover, we prepared PLGA nanoparticle-adsorbed PRRSV DNA vaccine and PEI-DNA complex, respectively. The results showed that all the experimental groups could significantly enhanced GP5-specific ELISA antibody, PRRSV-specific neutralizing antibody, IFN-γ level, as well as lymphocyte proliferation than the responses induced by pcDNA3.1-SynORF5. Among the experimental groups, PLGA-branched PEI nanoparticle induced the most efficient immune responses. The delivery systems and adjuvant in the research provide new path for the development of the new generation of vaccines against PRRSV.
Unprecedented short-term in vivo protection in mice with a single immunization of a DNA GP vaccine against heterologous mouse-adapted Ebola virus challenge
Authors: Patel Ami , Racine Trina , Reuschel Emma L. , Karuppiah Muthumani , Shedlock Devon , Yan Jian , Khan Amir , Tierney Kevin , Qiu Xiangguo , Kobinger Gary P , Sardesai Niranjan , Weiner David B 
Published: January 2023

Abstract
The recent Ebola outbreak in West Africa is a reminder that correlates of protection against filovirus infection are still not well understood. DNA vectors are a serology-independent platform that allow for vector re-administration with minimal side-effects. We designed a new micro-consensus DNA vaccine that expresses a Zaire Ebolavirus (EBOV) glycoprotein (GP) based on 2002–2008 EBOV outbreak strains. This novel glycoprotein is 3% distant from the GP expressed by the 1976 EBOV Mayinga outbreak strain. The optimized GP DNA vaccine was administered in mice by intramuscular injection followed by electroporation (IM-EP) and elicited strong total IgG antibody and T cell responses. To assess protection from challenge, we administered the GP DNA vaccine or pVax1 control (40ug) to BALB/c mice (n=10/group) by IM-EP at days 0, -7, -14, or -28 before lethal challenge (1000LD50) with a mouse-adapted EBOV Mayinga strain. Importantly we observed 100% protection against the lethal heterologous challenge when the DNA vaccine was administered 28 days before challenge illustrating the potency of this vaccine in this model. More importantly, we observed 100% protection and 90% protection when the DNA vaccine was administered at days -14 and day -7, respectively. Most surprising was the observation of 40% survival in the group that received vaccine at Day 0 (2–3 hours) before challenge. Rapid, short-term protection has been previously observed with a VSV-ZEBOVGP viral vector vaccine but never before with a DNA vaccine. The data suggests that protection in mice following administration of a heterologous DNA vaccine can be afforded by mechanisms independent solely of antibody titers. Ongoing studies are underway in NHPs to investigate these findings.