bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of an in vitro pre-clinical cellular assay to predict induction of Cytokine Storm by therapeutic antibodies. (125.16)
Authors: Strizheva Galina , Randhawa April , Mulligan Jenny , O’Day Christine 
Published: January 2023

Abstract
Cytokine storm (Hypercytokinemia/Cytokine Release Syndrome) is an acute immune reaction consisting of a positive feedback loop between cytokines and immune cells, resulting in severe inflammation and organ failure. A clinical trial of immunotherapeutic anti-CD28 antibody TGN1412 in 2006 demonstrated the importance of thorough pre-clinical screening and need for more sensitive assays to predict cytokine storm. In that study, 6 healthy volunteers suffered from hypercytokinemia at doses 500-fold less than the lowest dose tested in non-human primates (NHP), showing the failure of standard pre-clinical safety approaches of that time. We have developed an in vitro assay to evaluate antibodies for induction of cytokine storm. Frozen PBMC from humans and NHP are cultured with test articles bound to the cell-culture plate and assayed for cytokine production and cell proliferation. Anti-CD28 superagonist antibodies, which have been shown to stimulate human PBMCs using similar mechanisms as TGN1412, are used as a positive control. Human PBMC produced significantly more IL-2, IL-6, IFN-gamma, TNF-alpha, MIP-1-alpha and several other cytokines compared to isotype control or untreated cells, while PBMC from NHP were non-responsive. The increased production of key pro-inflammatory cytokines in response to stimulation with anti-CD28 antibodies is accompanied by increased PBMC proliferation, while no proliferation increase is detected in isotype-treated or unstimulated human PBMC.
Host F4/80+ macrophages are increased after transplantation of embryonic stem cell derived hematopoietic progenitors (126.10)
Authors: Thompson Heather , McLelland Bryce , Manilay Jennifer 
Published: January 2023

Abstract
Mice transplanted with embryonic stem cell (ESC)-derived hematopoietic progenitors (ES-HPs) historically have displayed low levels of engraftment. We hypothesized that the innate immune system was an obstacle to ES-HP engraftment. To investigate this, we derived ES-HPs in vitro by growing D3-ESCs (strain 129, CD45.2, H-2b) on the bone marrow (BM) stromal cell line, OP9. We transplanted purified CD41+ ES-HPs into sublethally irradiated NOD.SCID.γc knockout (NSG) mice hosts (CD45.1, H-2g7) which lack B, T, and NK cells, but have functional macrophages. Host and donor-derived hematopoiesis was observed. However, at Day 18 post-transplant, ES-HP recipients displayed enlarged spleens that were primarily composed of host macrophages. The absolute numbers of host-derived splenic F4/80+ macrophages were increased 1.81 fold compared to untransplanted controls and 1.7 fold compared to BM transplanted controls. In vitro phagocytosis assays confirmed that NSG macrophages specifically engulfed allogeneic ES-HPs, but not allogeneic adult BM cells, supporting our in vivo results. Taken together, these results suggest that host F4/80+ macrophages may pose a barrier to allogeneic ES-HP engraftment. Importantly, these data suggest that adult macrophages preferentially phagocytose ES-HPs, which could reveal a novel mechanism of rejection unique to ESC-derived cells. Experiments to evaluate the effects of host macrophage depletion on ES-HP engraftment in vivo are ongoing.
Multiphoton intravital microscopy visualization of skin graft rejection in real time (126.14)
Authors: Kuligowski Michael , Loh Yik , Kwan Rain , de St Groth Barbara 
Published: January 2023

Abstract
The current paradigm explaining organ graft rejection postulates that rejection is initiated by graft resident dendritic cells that reach the draining lymph nodes via lymphatics and activate naive alloreactive T cells. These newly activated T cells then migrate into the graft. However up to half the T cells recognizing allogeneic MHC in adult animals are already be of memory phenotype, and thus capable of direct entry into a graft site without prior activation in LNs. This is not the result of previous exposure to allo-Ag, but stems from the fact that the allo-MHC-specific and Ag-specific T cell receptor repertoires overlap each other. These pre-existing allo-reactive T cells represent a major barrier to organ transplantation. To understand the contributions of memory cells to skin graft rejection, we are using intravital multiphoton microscopy to track cellular interactions between memory cells, DCs and lymphatic and vascular endothelium during graft rejection and acceptance. Preliminary studies suggest that DCs from grafts are unable to exit the surgical site, most likely as a result of surgical disruption of lymphatic drainage from the transplanted skin. Further work will investigate trafficking of graft-reactive CD4 effector and Treg cells into the graft site.
Macrophage Inflammatory Protien-2/CXCR2 Blockade Attenuates Acute Graft-versus-Host Disease while Preserving Graft-versus-Leukemia Activity (126.16)
Authors: Park Minhwa , Cho Kyung-Ah , Woo So-Youn , Ryu Kyung-Ha 
Published: January 2023

Abstract
Graft-versus-host disease (GVHD) is the principal complication after allogeneic bone marrow transplantation (BMT). Strategies that effectively separate the beneficial GVL effects from GVHD toxicity are necessary for enhanced survival after BMT. GVHD mice were induced by infusion of both donor BM cells and spleen cells into BU-CY conditioned recipient female BALB/c mice. Tumor-bearing GVHD mice were also made adding A20 cells. A total of 31 cytokines and chemokines were measured in the liver, intestine, and skin in GVHD mice by Luminex Multiplex assay. Four cytokines, G-CSF, KC, MIP-2, and IL-23, were commonly unregulated in target organs at the initial GVHD stage and were reduced by neutralizing MIP2/CXCR2 axis ligation. CXCR2 blockade on donor T cells and systemic administration of anti-MIP-2 neutralizing antibodies to BMT recipients caused a significant decrease in both systemic GVHD and target organ histopathology. GVL responses (BCL2 expression or B220+/CD19+ cells in BM and PB) after MIP-2/CXCR2 blockade were largely preserved and mortality was significantly delayed compared to control allogeneic BMT recipients. These data suggest that blocking the MIP-2/CXCR2 axis represents a novel strategy to separate the toxicity of GVHD from the beneficial GVL effects after allogeneic BMT.
Plasmacytoid dendritic cell enrichment of bone marrow allografts enhances GvL activity by facilitating donor T cell engraftment and Th1 polarization (126.25)
Authors: Darlak Katarzyna , Wang Ying , Waller Edmund 
Published: January 2023

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) can cure patients with relapsed acute leukemia via a graft vs. leukemia (GvL) effect, but is complicated by graft vs. host disease (GvHD). Host dendritic cells (DC) following chemotherapy initiate GvHD in mouse models of allo-HSCT, but the role of donor DC is unclear. We have previously shown that transplanting purified plasmacytoid DC (pDC) along with HSC and T cells leads to Th1 donor T cell polarization and improved GvL activity. The goals of this study were: 1) To determine whether passive enrichment of pDC by depleting myeloid DC (mDC) from a bone marrow (BM) graft along with a fixed number of donor T cells would increase Th1 polarization and improve survival in leukemia-bearing mice, and 2) determine the role of IL-12 produced by donor pDC. Lethally irradiated B10.BR or FVB mice received BM and T cells from B6 or IL-12 KO B6 donors. Depletion of mDC from BM was achieved by FACS. Recipients of mDC-depleted BM had increased serum IFN-γ and increased donor T-cell proliferation and engraftment. Transplanting mDC-depleted BM enhanced survival without increasing GvHD in leukemia-bearing mice compared with undepleted BM. Enhanced proliferation and engraftment of donor T cells was not seen with mDC-depleted BM from IL-12 KO donors. These data show that pDC enrichment of BM allografts enhances donor T-cell Th1 polarization, engraftment, and GvL activity. The immune effects of mDC-depletion appear to be IL-12 dependent.
High affinity T cell receptor gives advantage to the homeostatic proliferation of naive CD8 T cells only under competitive pressure (126.28)
Authors: Ambros Tadeu , Zidan Abdel-Aziz , Pallin Maria , Montero Alberto , Lichtenheld Mathias , Díaz-Montero Marcela 
Published: January 2023

Abstract
The impact of T cell receptor (TCR) affinity during homeostatic proliferation has been demonstrated using naive CD8+ T cells with high and low affinity TCR derived from different precursors. In this study, we investigated the impact of TCR affinity using cells derived from the same precursor and selected under the same conditions, namely Pmel TCR transgenic (vβ13) naive CD8+ T cells, which respond to both high (hgp100) and low (mgp100) affinity peptides. Contrary to previous reports, we found that exposure of naive Pmel cells to either high or low-affinity peptide resulted in similar levels of homeostatic proliferation. However, in the presence of OT-1 cells and their cognate peptide (SINFKL), the levels of homeostatic proliferation of Pmel cells were higher after exposure to hgp100 than to mgp100 (1.85 vs 1.25 on day 8, 2.06 vs 1.20 on day 11, 2.57 vs 1.30 on day 15, 2.35 vs 0.89 on day 22). Interestingly, expression levels of memory markers (CD44lo to CD44hi, CD122lo to CD122hi and CD62Lhi to CD62Lhi) were similar under both circumstances. Overall, these results suggest that the difference seen in the effect of high-affinity TCR compared with low-affinity TCR on CD8+ cell expansion during homeostatic proliferation occurs only in the presence of competitive pressure. This difference may be balanced by a higher availability of low-affinity antigen, and thus may have important implications for vaccine design in the post-transplant setting.
The liver natural killer cells tolerate Alemtuzumab induction therapy in humans (126.3)
Authors: Ohira Masahiro , Nishida Seigo , Tekin Akin , Selvaggi Gennaro , Levi David , Tryphonopoulos Panaqiotis , Ruiz Phillip , Ricordi Camillo , Ohdan Hideki , Tzakis Andreas 
Published: January 2023

Abstract
Liver natural killer (NK) cells play a role in both graft acceptance and rejection in liver transplantation (LT). T cell depleting strategies have become an integral part of immunosuppressive regimens widely used for induction in transplant. Alemtuzumab (AL) is a recombinant humanized monoclonal antibody against human CD52. We observed early large granulocyte reconstitution in LT with AL. However, information is limited regarding the phenotype and function of NK cells in T cell depletion using AL. To investigate the role of NK cells in AL induction, we examine the phenotype and function of human NK cells derived from LT recipients and cadaveric donors. Absolute counts of T cells remained low for 6 months. In contrast, those of NK cells had emerged with larger number since early days after LT. Liver (n=7) contained high percentage of CD52-CD56+ NK cells (54.2 ± 9.0 % especially CD56 bright 81.2 ± 5.7 %), which suggested these NK cells could endure treatment of AL. It was statistically significant higher percentage than that of peripheral blood, spleen, and lymph nodes (25.1%, 28.4%, and 9.3 % respectively, P&lt;0.05). Donor liver NK cells had stronger cytotoxicity after IL-2 stimulation in the presence of AL. These data indicated that NK cells in peripheral blood were depleted but NK cells in liver remained alive and active in LT patient with AL induction. These results implicate that liver NK cells play important roles in transplant recipients with AL induction.
Role of syndecan-1 in TRAIL-induced apoptosis in myeloma cells (127.18)
Authors: Wu Yung-Hsuan , Yang Chen-Ying , Chien Wen-Li , Lin Kuo-I , Lai Ming-Zong 
Published: January 2023

Abstract
Syndecan-1 is a transmembrane heparin sulfate proteoglycan which functions as co-receptor for cell signaling and extracellular matrix in many types of malignancies. TNF-related apoptosis inducing ligand (TRAIL) is an antitumor agent which initiates apoptosis through the assembly of death receptor-inducing signaling complex (DISC) after ligand binding. However, role of syndecan-1 in TRAIL -mediated cell death is unknown. Here, we elucidated the pro-survival role of syndecan-1 in multiple myeloma cell lines. Knockdown of syndecan-1 in H929 and RPMI8226 cells increased TRAIL-induced apoptosis. The expression level of signaling molecules downstream of TRAIL was normal in knockdown cells. Although the binding and internalization ability of TRAIL were still normal, an enhanced formation of DISC and caspase-8 activation were detected in knockdown cells. In addition, TRAIL-induced oligomerization of DR4 was enhanced in syndecan-1-knockdwon cells. Thus, syndecan-1 may regulate TRAIL-mediated cell death in the clustering of death receptors in multiple myeloma cells
The upregulation of the inhibitory receptors BTLA and PD-1 defines a novel subset of dysfunctional tumor antigen-specific CD8+ T cells (127.19)
Authors: Fourcade Julien , Sun Zhaojun , Pagliano Ornella , Guillaume Philippe , Luescher Immanuel , Sander Cindy , Kirkwood John , Olive Daniel , Kuchroo Vijay , Zarour Hassane 
Published: January 2023

Abstract
Cytotoxic T cells present at the periphery or at tumor sites recognize tumor epitopes presented by melanoma cells but fail to induce tumor rejection in melanoma patients. Inhibitory receptors including PD-1 and Tim-3 expressed by spontaneous CD8+ T cells directed against the cancer-germline antigen NY-ESO-1 play a role in promoting tumor-induced T cell dysfunction. In the present study, we show that BTLA is upregulated on spontaneous NY-ESO-1-specific CD8+ T cells and that BTLA-expressing PD-1+Tim-3- CD8+ T cells represent the largest subset of NY-ESO-1-specific CD8+ T cells in melanoma patients. These cells were partially dysfunctional, producing less IFN-γ than BTLA- T cells, but more IFN-γ, TNF and IL-2 than the highly dysfunctional subset expressing all three receptors. Unlike PD-1, BTLA expression by NY-ESO-1-specific CD8+ T cells does not increase with higher T cell dysfunction or upon cognate antigen stimulation, suggesting that BTLA upregulation occurs independently of functional exhaustion driven by high antigen load. Finally, BTLA blockade adds to PD-1 and Tim-3 blockades, enhancing NY-ESO-1-specific CD8+ T cell expansion, proliferation and cytokine production. Collectively, our findings support the targeting of BTLA, PD-1 and Tim-3 pathways to reverse tumor-induced T cell dysfunction in patients with advanced melanoma.
Effects of a multi-targeted receptor tyrosine kinase inhibitor on radiosentization of head and neck squamous cell carcinoma. (127.21)
Authors: Mirshahidi Saied , Hsu Heng- Wei , Chen Chien-Shing 
Published: January 2023

Abstract
Targeted biological therapies that selectively interfere with cancer cell growth signals may improve patient survival by enhancing the effects of radiation, with the added benefit of reduced systemic toxicity. Head and Neck Squamous Cell Carcinoma (HNSCC) is the most common epithelial malignancy arising in the upper aerodigestive tract. It is the sixth most common cancer worldwide. Signal transducers and activators of transcription 3 (STAT3), is present in a number of different cancer cells including head and neck cancer and is related to chemoresistance and radioresistance, therefore is a potential novel molecular target for therapy to improve survival of the patients. In the present study, we report that linifanib (ABT-869), is a novel multi-targeted receptor tyrosine kinase (RTK) inhibitor of members of vascular endothelial growth factor (VEGF) and platelet derived growth factor (PDGF) receptor families, has anti-proliferative effects in HNSCC cell lines in a dose dependent manner. It inhibits STAT3 signaling and related proteins and arrests cell cycle at G2/M phase, induces apoptosis, inhibits colony formation efficiency of the cells. In addition, when this multi-targeted receptor tyrosine kinase inhibitor is combined with radiation, significant antitumor effects are achieved in the cell lines. Thus, it may provide a therapeutic strategy and improves efficacy of radiation against HNSCC.
Natural CXCR3+ FOXP3+ Treg selectively accumulate in human ovarian carcinomas to control type I immunity (127.27)
Authors: Raffin Caroline , Redjimi Nassima , Odunsi Kunle , Valmori Danila , Ayyoub Maha 
Published: January 2023

Abstract
Anti-tumor type I T cell responses are critical to control cancer but their efficacy is limited by immunoregulatory mechanisms, including the action of immunosuppressive FOXP3+ Treg. Recent studies have revealed a significant heterogeneity in FOXP3+ Treg, that, similar to conventional CD4+ T cells, include subsets co-expressing lineage-specific transcription factors, that specifically control over the corresponding conventional effector subset. FOXP3+ Treg expressing CXCR3 have been recently described as a subset specialized in the control of type I T cell responses in vivo. We found that CXCR3+ Treg are highly enriched in ovarian carcinomas (OC), particularly in solid tumor masses, where they represent the majority of Treg. CXCR3+ Treg in OC express T-bet but do not secrete IFN-γ ex vivo and suppress proliferation and IFN-γ secretion of T effectors. OC CXCR3+ Treg are Helios+, suggesting that they originate from natural rather than from adaptive Treg, In support of this, CXCR3+ T-bet+ Helios+ IFN-γ- Treg are efficiently generated upon stimulation of natural naïve Treg under TH1 polarizing conditions. Finally, we show that the presence of CXCR3+ Treg is directly correlated with that of CXCR3+ T effectors and is consistent with expression of CXCR3 ligands at OC sites. Together, our findings support the concept that natural CXCR3+ Treg selectively accumulate in OC tumors to control type I T cell responses, which results in the limitation of efficient anti-tumor immunity.
IL-12 and Type I IFN differentially program CD8 T cells for PD-1 expression levels and tumor control (127.32)
Authors: Heltemes Harris Lynn , Gerner Michael , Mescher Matthew 
Published: January 2023

Abstract
Naïve CD8 T cells proliferate in response to signals through TCR and CD28, but require IL-12 or Type I IFN to undergo differentiation and develop optimal effector functions. CTLs generated in vitro in response to IL-12 or IFN-alpha displayed comparable cytolytic activity and IFN-gamma production at day 3. Despite similar day 3 activity, IL-12-stimulated cells were much more effective in controlling tumor growth in an adoptive immunotherapy model. IL-12-stimulated cells maintained high numbers and function in the tumors, while IFN-alpha-stimulated cells declined in number and became exhausted. Consistent with this, IFN-alpha-stimulated cells expressed significantly higher levels of the PD-1 inhibitory receptor than did IL-12-stimulated cells. The efficacy of IFN-alpha-stimulated CTL in adoptive immunotherapy was strongly increased, and became comparable to that of IL-12-stimulated cells when a blocking antibody specific for PD-L1, the ligand of PD-1, was administered. Thus, IL-12 and IFN-alpha differentially program CD8 T cells to re-express distinct levels of PD-1 when the cells re-encounter antigen, resulting in IL-12-stimulated cells being less susceptible to exhaustion in the face of sustained tumor antigen.
Characterization of human B cells and macrophages infiltrating ovarian cancers. (127.33)
Authors: Nunez-Cruz Selene , Walter Marc , Danet-Desnoyers Gwenn , Scholler Nathalie 
Published: January 2023

Abstract
Suitable tumor xenograft models represent a significant improvement to the understanding of ovarian cancer. However phenotype and function of tumor-associated lymphocytes and macrophages in different tumor sites has not been reported. We characterized the ovarian tumor microenvironment in NSG mice reconstituted with human immune system using cord blood-derived CD34+ cells. Mice were injected with OVcar5 human ovarian tumor cells orthotopically (io), subcutaneously (sc) or both (io/sc) 12-16 weeks post CD34-transplant. Our findings indicated that 4 weeks after implantation, all human tumors were infiltrated with B cells, T cells and macrophages, regardless of the tumor site. However, sc tumors contained proportionally more B cells and macrophages than io tumors. Macrophages infiltrating sc and io tumors exhibited an IL10high immunosuppresive phenotype. Io macrophages expressed higher levels of the Fc receptor of high affinity (FcgRI) and of low affinity (FcgRII) and mannose receptor compared to macrophages infiltrating sc or io/sc tumors. Finally, io tumors were infiltrated by more B cells than all other tumors. B cells from tumor-bearing mice presented an altered phenotype compared to non tumor-bearing animals, characterized by the upregulation of plasma and activation markers and IgM and IgG production. Our results suggested that the location of the tumor implantation impacts the phenotype and function of human tumor infiltrate and may influence the tumor development.
Tumor-induced myeloid-derived suppressor cell function is independent of IFNγ and IL-4Rα (127.37)
Authors: Sinha Pratima , Parker Katherine , Ostrand-Rosenberg Suzanne 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSC) are present in most cancer patients and experimental animals where they exert a profound immune suppression and are a significant obstacle to immunotherapy. IFNγ and IL-4Rα have been implicated as essential molecules for MDSC development and immunosuppressive function. If IFNγ and IL-4Rα are critical regulators of MDSC, then they are potential targets for preventing MDSC accumulation or inhibiting MDSC function. Because data supporting a role for IFNγ and IL-4Rα are not definitive, we have examined tumor-induced MDSC from IFNγ-deficient, IFNγR-deficient, and IL-4Rα-deficient mice. Although MDSC express functional IFNγR and IL-4Rα and signal via the STAT1 and STAT6 pathways, respectively, neither IFNγ nor IL-4Rα impacts the phenotype, accumulation, or T cell suppressive potency of MDSC, although IFNγ and IL-4Rα modestly alter macrophage -induced MDSC production of IL-10. Therefore, neither IFNγ nor IL-4Rα is a key regulator of MDSC and targeting these molecules is unlikely to significantly alter MDSC function.
Resveratrol abrogates tumor-evoked Bregs conversion by targeting PPAR-alpha pathway (127.41)
Authors: Lee-Chang Catalina , Wejksza Katarzyna , Bodogai Monica , Malchinkhuu Enkhol , Ghosh Paritosh , Martin-Montalvo Sanchez Alejandro , de Cabo Rafael , Biragyn Arya 
Published: January 2023

Abstract
We recently reported that breast cancer cells induce the generation of a regulatory B-cell subset (tumor-evoked Bregs, tBregs) from naïve B cells to promote cancer escape and lung metastasis. One of the mechanisms involved is the ability of tBregs to promote Treg expansion. Thus, aiming to understand this process we screened several compounds that could block the generation of tBregs. We found that resveratrol (RSV), a natural polyphenol with chemopreventice properties, specifically blocked the ability of cancer cells to convert B cells into tBregs. This blockage results in the inability of tBregs to suppress T-cell proliferation and promote tumor growth and lung metastasis. The molecular mechanism of this process is that RSV targeted and inactivated PPAR-alpha, a nuclear receptor implicated in several processes such as lipid metabolism, proliferation or differentiation. Thus, our data show the importance of PPAR-alpha in tBreg generation, and one mechanism of the anti-tumor properties of RSV is the inactivation of this pathway in tBregs. This work was entirely supported by the Intramural Research Program of the National Institutes of Health.