bluuebunny/crossref_metadata_2025_split huggingface dataset:
Suppressive effects of fucoidan-agaricus mix (CUA-fucoidan) on angiogenesis and tumor growth in B16 melanoma-bearing mice
Authors: Miyazaki Yoshiyuki , Bak Juneha , Nakano Hayato , Takeuchi Shugo , Takeuchi Hideaki , Tachikawa Daisuke 
Published: January 2023

Abstract
Fucoidan is a series of natural sulfated polysaccharides contained in brown sea algae. In previous studies, we proved that fucoidans derived from Cladosiphon okamuranus and Undaria pinnatifida effectively augmented anti-tumor immunity in combination with an Agaricus blazei mycelium extract. In this study, we evaluated dietary effects of the mixture consist of the two high-molecular weights fucoidan and the agaricus extract (named CUA-fucoidan) on tumor angiogenesis. For that purpose, C57BL/6 mice were subcutaneously injected basement membrane matrix (Geltrex) enclosing 5×104 mitomycin C-treated B16 melanoma cells or 500 ng/mL vascular endothelial growth factor (VEGF), and fed 2% CUA-fucoidan containing diet for 7 days before and after the matrix inoculation. The accumulation of hemoglobin and CD31-positive cells in the B16-enclosed Geltrex was lower in the CUA-fucoidan-fed mice than in the control mice, whereas the hemoglobin contents in the VEGF-enclosed Geltrex were not affected by the CUA-fucoidan feeding. These results suggested that CUA-fucoidan was effective in suppressing tumor angiogenesis, but did not inhibit the physiological action of VEGF. Rather, it was guessed that CUA-fucoidan suppressed VEGF production because the expression of VEGF mRNA was reduced in the matrix resected from CUA-fucoidan-fed mice. In this context, the infiltration of CD206-positive macrophages, which known to produce VEGF, were declined by the CUA-fucoidan feeding. Furthermore, CUA-fucoidan decreased accumulation of Treg in the B16-included Geltrex, and delayed B16 tumor growth. In conclusion, CUA-fucoidan was expected to be effective in inhibition of tumor angiogenesis by improving immunosuppressive tumor microenvironments.
Development of a Humanized 3D Tumor Spheroid Models and Discovery of Effective Immunotherapy Strategies Against Resistant Microsatellite Stable Colorectal Cancers
Authors: Torres Adorno Angie M , Fong Julie , Grein Jeff , Blumenschein Wendy , Cheng Danye , Porth Katherine , Yang Peng , Yearley Jennifer , Beebe Amy , Joyce-Shaikh Barbara 
Published: January 2023

Abstract
Despite the current success of immunotherapies, only microsatellite instable (MSI) Colorectal Cancer (CRC) typically respond to T cell-driven immunotherapies while microsatellite stable (MSS) CRC displays poor response. Our goal is to identify resistance mechanisms in MSS-CRC to uncover novel therapeutic targets. Current syngeneic CRC models are largely immunogenic and lack complex structure. Therefore, we developed novel 3D humanized CRC mouse models which form complex human-like tumors which are typically absent in syngeneic systems. We investigated molecular profiles of human MSI-versus MSS-CRC tumor xenografts following Keytruda immunotherapy and identified an immunosuppressive profile in MSS models that was not observed in MSI-CRC. We evaluated the therapeutic potential of blocking inflammation in combination with other immuno-modulatory therapies in both cold and hot CRC models and identified a significant regression of tumor burden in MSS-CRC mice. Our studies provide a novel CRC in vivo model and a unique opportunity to better define how immune cells interact in human MSS- and MSI-CRC microenvironments, to enable clinical therapeutic implementation.
Immunological responses through miRNA signatures in GBM plasma extracellular vesicles from patients receiving experimental immunotherapy
Authors: Garcia Luz Milbeth Cumba , Dehankar Mrunal K , Nair Asha A , Dietz Allan B , Parney Ian F 
Published: January 2023

Abstract
Glioblastoma (GBM) patients have a median survival of 15 months despite aggressive treatment. Treatment-related pseudo-progression confounds outcome assessment by MRI, particularly in patients receiving immunotherapy. Extracellular vesicles (EVs) contain tumor-specific miRNA that could serve as a liquid biopsy to distinguish true progression from treatment-related pseudo-progression. Small plasma EVs were isolated by serial density gradient ultracentrifugation from 20 newly-diagnosed GBM patients enrolled in a clinical trial of allogeneic tumor lysate-pulsed autologous dendritic cell (DC) vaccination. Short non-coding RNA sequencing was performed for each patient at three time points (TP): pre-vaccine (TP1), post-initial vaccine (TP2), and at end of treatment (TP3). Ingenuity Pathway Analysis from 31 differentially expressed miRNAs across time points (p-value &lt;0.05, |logFC|&gt;1) revealed that glioma-related signaling pathways previously seen in small EVs from glioma patients compared to normal donors were replicated when comparing patients at the beginning of the treatment (TP1-vs-TP2) and between early to late time points (TP1-vs-TP3). In addition, pathways corresponding to immunological responses typically seen in GBM patients undergoing treatment, such as IL-10, IL-6, and Toll-like Receptor (TLR) signaling, were exclusively found after the initial vaccine towards the end of treatment (TP2-vs-TP3). In conclusion, signaling pathways derived from differentially expressed miRNAs across time points suggest that as patients progress through treatment, consistent differences in plasma small EVs miRNA expression profile and immunological responses could be utilized as predictors of treatment response.
Reprograming of Tumor-Associated Macrophages in Human BrafV600EMutant Melanoma
Authors: Santiesteban Gretel Maria Torres , Bhandari Rajan , Yang Heetaek , Spezza Paul , Shabaneh Tamer Basel , Liby Karen , Turk Mary Jo , Pioli Patricia A 
Published: January 2023

Abstract
Melanoma tumors are highly immunogenic, making them an attractive target for immunotherapy. However, many patients do not mount robust clinical responses to targeted therapies, which is attributable, at least in part, to suppression of immune responses by tumor-associated macrophages (TAMs) in the tumor microenvironment (TME). Using a human in vitro culture system, we have shown that the synthetic triterpenoid CDDO-Me enhances immune activation by reprogramming macrophages from immuno-suppressive to immuno-stimulatory. CDDO-Me significantly reduced CCL2, VEGF and IL-6 secretion by macrophages and inhibited surface expression of CD163, a marker associated with poor clinical outcomes. Furthermore, CDDO-Me mediated both contact-dependent and independent effects on macrophage activation in tri-cultures of macrophages with activated autologous T cells and BRAFV600E mutant melanoma cells. Signaling pathway activation was interrogated to identify the potential mechanism by which CDDO-Me attenuates the pro-tumorigenic macrophage activation profile. Our results demonstrated that CDDO-Me inhibited STAT3 phosphorylation, which is a known regulator of TAM activation. Given these results, we hypothesize that, in addition to mediating direct anti-tumorigenic effects, CDDO-Me may also enhance the efficacy of additional immunotherapies, including BRAF inhibition. Collectively, our studies suggest that the redirection of immuno-suppressive myeloid cell activation may provide both a direct means of inhibiting melanoma growth and may enhance the efficacy of additional targeted immunotherapies through relief of immune suppression in the TME.
ST8Sia6 Overexpression Accelerates Tumorigenesis in a Siglec E Dependent Manner Through Reshaping the Tumor Immune Microenvironment
Authors: Friedman David J , Shapiro Michael J , Rajcula Matthew , McCue Shaylene , Khazaie Khashayarsha , Dong Haidong , Shapiro Virginia Smith 
Published: January 2023

Abstract
Checkpoint blockade has improved patient outcome through reinvigorating T cells against cancer. Despite this success, many patients don’t respond, leaving them with few options. Tumors utilize many immunosuppressive mechanisms, including altering the function of innate immune cells. Cancer cells can promote differentiation of macrophages into tolerogenic “M2-like” macrophages, which support tumor growth. Macrophage differentiation by cancer cells is influenced by many receptors, including Siglec-E. Siglec-E binds to α2,8-linked disialic acids, generated by the enzyme ST8Sia6, on O-linked glycoproteins, and functions to inhibit activation of innate immune cells. Cancer patients with high expression of ST8Sia6 exhibit reduced survival. Therefore, ST8Sia6 overexpressing tumors may engage Siglec-E, leading to immune suppression and accelerating tumor growth. Mice injected with ST8Sia6 overexpressing MC38 and B16-F10 tumors had increased tumor burden, reduced survival and an increase M2 macrophage polarization. The acceleration in tumor growth was abrogated when injected into Siglec-E knockout mice. Thus, the primary function of ST8Sia6 in tumors is an extrinsic alteration of the immune response, rather than an intrinsic effect on cell growth. Similarly, in a spontaneous colon cancer model, ST8Sia6 overexpression in a murine colon cancer model accelerated the development of tumors, decreasing survival to 2.5 months instead of 6 months. The overexpression of ST8Sia6 on tumors leads to increased Arg-1 expression by CD11b+ CD11c+Ly6C−Ly6G− cells, which suppresses the immune response. Therefore, ST8Sia6 expression in tumors inhibits the immune response to promote tumor growth in a Siglec-E dependent manner.
Isolation of human CD45+ leukocytes from tissues and human tumor xenografts in humanized mice
Authors: Antignano Frann , Posarac Vesna , Ewen Catherine L , Jovanovic Vida K , Liang Alice , Woodside Steven M , Kokaji Andy I , Eaves Allen C , Thomas Terry E 
Published: January 2023

Abstract
The study of immune cell function in non-lymphoid tissue and tumors promises to elucidate novel strategies to treat immune disorders, infectious diseases, and cancer. To address the challenge of isolating leukocytes from complex and variable tissues and tumors, we have developed a new protocol to isolate particle-free, human CD45+ leukocytes. Using the EasySep™ Release Human CD45 Positive Selection Kit, leukocytes are labeled with antibody complexes linked to magnetic particles and separated using an EasySep™ magnet. The magnetic particles are then removed from the desired cells by resuspension in EasySep™ Release Buffer and a final magnetic separation.
To assess performance, NRG-3GS mice were first engrafted with human CD34+ cells followed by xenotransplant with human breast (MDA-MB-231) or ovarian (SKOV3) cancer cell lines. In humanized mouse lungs, bone marrow and spleen, the starting and isolated human CD45+ frequency ranges were 6.0 – 57.2% and 90.9 – 99.4%, respectively (n = 3). Starting with human tumor xenografts, tumor infiltrating leukocytes were enriched from a starting range of 0.4 – 18.0% to 76.6 – 92.7% (n = 4). The final immune cell frequencies are representative of the starting population, and further separation of immune subsets can be achieved with additional downstream isolation.
Humanized mouse models of clinical disease are instrumental in furthering our understanding of complex mechanisms of disease progression and resolution. This new kit for the isolation of human immune cells from tissues and tumors will facilitate further examination of the roles of immunity in disease and the evaluation of immune-based treatment strategies.
Bone microenvironment-suppressed T cells increase osteoclast formation and breast cancer bone metastases
Authors: Arellano Danna L , Corral-Avila Juan A , Drescher Florian , Verdugo Andrea , Jimenez Samanta , Olvera-Rodriguez Felipe , Juarez Patricia , Fournier Pierrick 
Published: January 2023

Abstract
Bone metastases is a frequent (&gt;75%) complication of advanced breast cancer (BCa). BCa cells in bone are supported by cytokines released during the osteoclastic resorption, and remain untreatable. T cells during immunotherapy could turn against BCa cells in bone but could also increase osteoclasts and bone metastases.
Comparing the development of bone metastases from 4T1 BCa cells between Balb/C and SCID or T cell-depleted mice, the absence of T cells decreased bone metastases (−72% and −38%, respectively), while orthotopic tumors were increased. Histology confirmed that the osteoclast number was decreased at the tumor-bone interface of mice lacking T cells. Ex vivo addition of non-activated T cells increased osteoclast formation, and flow cytometry confirmed that &gt;85% of T cells in bone metastases were not expressing activation markers.
In sharp contrast, activated T cells were potent inhibitors of osteoclast formation, which could be beneficial for the treatment of patients. Although activated T cells expressed the anti-osteoclastic Ifng and Il4, their effect was not reversed by neutralizing antibodies, or due to a bystander effect by cytotoxic T cells. Thus, we sought to activate T cells of bone metastases. However, it was not possible to activate T cells in bone marrow cultures ex vivo. This could be due to an increase of MDSCs in 4T1 bone metastases, including M-MDSCs that were PD-L1+ (&gt;85%) and could suppress PD-1+ T cells (&gt;70%) in bone.
Our results suggest that while T cells under the influence of the bone microenvironment promote BCa bone metastases, immunotherapy-activated T cells could suppress bone resorption and eliminate BCa cells in patients with bone metastases.
Protein Kinase D: A new target in the fight against flu?
Authors: Veazey Janelle , Chapman Timothy J , Smyth Timothy R , Hillman Sara E , Eliseeva Sophia I , Georas Steve N 
Published: January 2023

Abstract

Summary
Antivirals targeting host proteins may slow development of viral resistance and work against multiple viruses. The serine/threonine kinase protein kinase D (PKD) is one promising target as others have shown inhibiting PKD restricts rhinovirus and hepatitis in cell culture. We show inhibiting PKD restricts Influenza A virus (IAV) in cell culture and in mice.

Methods
The competitive PKD inhibitor, CRT, was given to A549, 16HBE cells and mice, prior to or during stimulation with dsRNA or IAV.
Pulmonary barrier integrity was quantified by assaying total protein leak into the lumen (inside/out leak), and loss of 4kDa FITC-dextran out of the lumen (outside/in leak).
Inflammatory cytokine levels were determined via ELISA/multiplex and mRNA levels via RT-PCR. Transcription factor activity was assayed via luciferase reporter system.
Viral load was determined via RT-PCR of viral M protein. For in vivo IAV experiments, mice were given CRT/vehicle post-infection with lethal dose IAV (PR/8) and morbidity assessed by weight loss.

Results

Conclusion
PKD inhibition promotes barrier integrity, lowers transcription of potentially pathogenic pro-inflammatory cytokines, and restricts viral replication, highlighting the potential of CRT as a novel anti-viral therapeutic.
Immunogenic potency of engineered exosomes for prevention of respiratory syncytial virus
Authors: HONG SUYEON , Ruan Shaobo , Kubota Pamela Gamero , He Mei , McGill Jodi L 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract disease in infants and young children. Bovine RSV is a common cause of respiratory disease in young cattle and is a useful model to study RSV infection. There is currently no vaccine for RSV treatment in humans, and the vaccines available for BRSV have limited efficacy in the field. Exosomes (30–150nm) are cell-derived extracellular vesicles that play critical roles in cellular communication by carrying various molecules. Since exosomes secreted by virus or tumor infected cells can deliver cell-specific antigens and proteins to immune cells, immunogenic exosomes have emerged as a new vaccine platform for prevention of viral infections or therapy for cancer treatment. Here, dendritic-cell derived exosomes were engineered to present peptides from the RSV M, NS and L proteins, purified from PDMS microfluidic systems and were evaluated as an efficacious surface engineered antigenic exosome vaccine against RSV and BRSV in mice. The immunogenicity of the exosomes was confirmed in vitro by measuring their capacity to induce antigen-specific T cell clonal proliferation and IFNγ production. The in vivo immunogenicity and efficacy of the engineered exosomes was investigated by immunizing animals subcutaneously with escalating doses of engineered exosomes and measuring antigen-specific T cell responses in the spleen and lungs, and assessing morbidity and mortality. Collectively, dendritic-cell derived surface engineered antigenic exosomes have the capacity to elicit antigen-specific CD8 T cell activation in vitro and in vivo, and the potential to be developed as a vaccine platform to prevent RSV and other viral infections in humans and animals.
Dissecting human, ferret, and murine antibody and B cell responses to generate a broadly protective synthetic influenza virus hemagglutinin
Authors: Kim Hyunsuh , DeBeauchamp Jennifer , Stigger Evelyn , Ellebedy Ali , Rubrum Adam , El-Shesheny Rabeh , Crumpton Jeri , Kackos Christina , Thomas Paul G , Caniza Miguela , DeVincenzo John , Ahmed Rafi , Webby Richard J 
Published: January 2023

Abstract
A major challenge in developing improved influenza vaccines is the identification of broadly reactive and protective epitopes. While it is generally believed that the specificity of the antibody response to hemagglutinin (HA) is relatively narrow, we and others have identified individuals that mount a cross-reactive neutralizing antibody response after natural infection. Using plasmablasts from one such individual we generated broadly cross-reactive HA-specific monoclonal antibodies (mAbs). Six of these mAbs bound to the globular head of the HA and one to the stalk region; all provided broad protection against multiple subtypes of influenza A viruses including zoonotic strains in passively immunized mice. Using a peptide ELISA approach, we identified the binding regions of each mAb and subsequently immunized mice with these peptides. Immunized mice showed less weight loss and significantly improved survival against challenges with multiple subtypes as compared with control mice. We next used a similar mapping approach with broadly reactive B cells from mice and ferrets. Based on all of the collected data we generated three different composite HA antigens (HF1, HF2 and HF3) that contained a different mix of broadly reactive epitopes in one native HA structure. Mice immunized with the HFs were protected from multiple challenges, including lethal doses of H1, H3, H5, H6, H7 and H9 viruses. We also showed the HFs protected ferrets from H1, H3, and H9 viruses. Overall, by combining data generated from an examination of B cell and antibody responses following infection and/or vaccination in humans, mice, and ferrets we were able to construct a composite HA that was able to induce broad protection against multiple subtypes of influenza virus.
The effect of SMAC mimetics on human T cell activation
Authors: Murray Susan E , Ligman Brittany , Burton Ashley , Kearney Claire 
Published: January 2023

Abstract
Second mitochondria-derived activator of caspase (SMAC) mimetics are small molecule drugs currently in cancer clinical trials. They are thought to act by antagonizing inhibitors of apoptosis (IAPs), which are often overexpressed in malignancies. However, in leukocytes, IAPs inhibit cell activation via antagonizing non-canonical NF-κB. This suggests that, paradoxically, while SMAC mimetics can induce apoptosis in cancer cells, they may enhance activation in T cells. The aim of this project is to determine whether human T cell proliferation and effector function are affected by SMAC mimetics.
We previously showed that non-canonical NF-κB plays an essential intrinsic role in T cell activation in mice. Here, we show that the SMAC mimetic TL32711 (Birinapant) activates non-canonical NF-κB in primary human T cells. Doses of SMAC mimetics that induce 100% apoptosis in the breast cancer cell line, MDA-MB-231, do not affect T cell viability. Birinapant dose-dependently increases IL-2 secretion, but the effects of SMAC mimetics on T cell proliferation and Th1 cytokine production are complex and depend on availability of costimulation and accessory cells.
Endogenous T cells can combat cancer if properly stimulated, a fact underlined by the efficacy of checkpoint blockade. Thus, understanding the effect of SMAC mimetics on T cell responses is a critical consideration in using SMAC mimetics to treat cancer and for designing combination therapies.
Targeting microbial-specific gamma/delta T-cell subset for vaccine development
Authors: Chen Zheng , Shen Ling 
Published: January 2023

Abstract
Tuberculosis (TB), caused by M. tuberculosis (Mtb), remains the top killer among infectious diseases. The current TB vaccine, Bacille Calmette–Guérin (BCG), only protects young children from severe disseminated TB, but not effectively protects against pulmonary TB in adults. Improved TB vaccine or vaccination approach is needed for effective global prevention against TB. Our decade-long mechanistic studies in nonhuman primates (NHP) employed 2 innovative “gain-of-function” manipulations to generate Vγ2Vδ2 T effector cells in vivo for definition of their function and immunity against high-dose TB. These seminal studies established that HMBPP-specific Vγ2Vδ2 T effector cells are fast-acting, multi-functional and protective against high-dose TB in NHP. These novel findings prompted us to perform creative immunization of Vγ2Vδ2 T subset using our innovative vaccine vector Listeria ΔactA prfA*. Respiratory vaccination of macaques with an HMBPP-producing attenuated LM ΔactA prfA*, but not the control, caused prolonged expansion of HMBPP-specific Vγ2Vδ2 T cells in pulmonary and circulating compartments. After pulmonary Mtb challenge, macaques vaccinated with LM ΔactA prfA* exhibited rapid memory-like response of Vγ2Vδ2+ Th1 cells. The selective immunization of Vγ2Vδ2 T cells contained Mtb infection/dissemination and reduced lung TB pathology, enhancing the ability of tissue-resident Vγ2Vδ2 T cells to inhibit Mtb growth in macrophages. Thus, selective immunization of Vγ2Vδ2 T cells elicits fast-acting/durable memory-like responses, providing an approach to creating more effective TB vaccines.
Late-rising CD4 T cells show dramatic control of cytomegalovirus persistence
Authors: Picarda Gaelle Genevieve Alice , Brunel Simon , Ghosh Raima , Gupta Ankan , Greenbaum Jason , Seumois Gregory , Croft Michael , Vijayanand Pandurangan , Benedict Chris A 
Published: January 2023

Abstract
Conventional CD4 T cells typically expand and establish memory pools within the first few weeks of acute viral infection. However, cytomegalovirus (CMV) exhibits an extended persistent replication phase followed by lifelong latency, suggesting the induction, phenotype and/or function of protective memory CD4 T cells may differ. We show that during CMV infection in mice, CD4 T cells recognizing an epitope derived from the viral M09 protein develop long after conventional memory T cells have already formed. These cells are unique compared to their conventional memory counterparts; showing high IFNγ and IL-2 production at times of viral persistence. Ablating these CD4 T cells by mutating the M09 genomic epitope, or inducing them through vaccination, revealed them far superior at resolving persistent replication. RNAseq data reveal that these ‘late-rising’ and conventional CD4 T cells differ dramatically in their transcriptional signatures, and that M09 cells show a uniquely restricted T cell receptor repertoire. We believe these results may help instruct CMV vaccine and cellular immunotherapy approaches in people, and will also provide a blueprint for determining whether similar late-rising CD4 T cells exist in other chronic virus infections.