bluuebunny/crossref_metadata_2025_split huggingface dataset:
Acute GVHD in liver and lungs is mediated by phenotypically-analogous but transcriptionally unique T cells.
Authors: Omdahl Kayleigh I , Bermea Rene , McGuckin Connor , Kaminski Jim , Lane Jennifer , Tkachev Victor , Kean Leslie 
Published: January 2023

Abstract
During acute Graft-versus-Host Disease (aGVHD), donor T cells invade host tissues and cause multi-organ damage. However, the organ-specific drivers of pathogenic alloimmunity are still poorly understood. To uncover both the universal and organ-specific mechanisms driving T cell-mediated damage in target tissues during aGVHD, we analyzed the flow cytometric phenotype as well as transcriptomic signatures of T cells in healthy control animals, in autologous hematopoietic cell transplant (HCT) controls, and during aGVHD after allogeneic HCT, using our established non-human primate aGVHD model. In this study we focused on 2 aGVHD target organs: the liver and lungs. Using both supervised and unsupervised flow cytometric analysis, we discovered that, during aGVHD, both liver- and lung-infiltrating T cells demonstrated similar phenotypes dominated by mature effector-memory CD8 T cells. In contrast, transcriptomic analysis revealed organ-specific gene expression programming. Through Weighted Gene Co-expression Network Analysis (WGCNA), differential expression (DE) analysis, and immune pathway analysis, we identified that the lung aGVHD T cell transcriptomic program was specifically associated with extracellular matrix-remodeling and chemotaxis pathways. In contrast, in liver aGVHD, we found enrichment for metabolic stress responses and cytokine signaling pathways. These results suggest that at the transcriptomic level, T cells infiltrating different aGVHD-target organs are uniquely programmed and could be amenable to organ-specific therapeutic targeting.
Supported by grants from the NIH (F31HL1562880)
PD-1H/VISTA expressed on host myeloid cells regulates acute graft-versus-host disease (aGVHD)
Authors: Hu Qianni , Hedgepath Carly , Hong Junshik , Huh Won Jae , Han Xue , Vesely Matthew , Chen Lieping , Kim Tae Kon 
Published: January 2023

Abstract
Programmed Death-1 Homolog (PD-1H, also known as VISTA) is a co-inhibitory molecule of the immunoglobulin superfamily and broadly found in hematopoietic cells. We previously demonstrated that PD-1H agonists prevent aGVHD in a murine MHC mismatched bone marrow (BM) transplantation model. aGVHD occurs when donor T cells are primed and activated by host antigen-presenting cells (APCs). PD-1H agonism suppressed donor allogeneic T cell activation leading to the suppression of aGVHD. However, it remains unknown whether PD-1H agonists also suppress host myeloid APCs, in which PD-1H is highly expressed. To test this, we adoptively transferred wildtype (WT) C57B/L6 (B6) BM into either PD-1H knock-out (KO) or WT BALB/c mice as hosts. PD-1H KO host mice had more severe aGVHD clinical scores and a worse survival rate when compared to WT host mice. To determine whether PD-1H agonism impacts host immune cells, we transferred B6 PD-1H KO BM into BALB/c WT host mice and treated host recipients with or without PD-1H agonists. Consistent with prior data, host mice treated with PD-1H agonists suppressed aGVHD and extended survival compared with isotype control treated mice. To test which cell subsets expressing PD-1H in host mice regulate aGVHD, we transferred WT BALB/c BM into myeloid lineage specific KO of PD-1H or WT littermate hosts. Interestingly, the myeloid lineage specific KO mice host had significantly worse survival rates and worse aGVHD symptoms compared with WT littermates. Our studies indicate that PD-1H expression on host myeloid cells regulates aGVHD in addition to donor T cell PD-1H. Finding new mechanisms of PD-1H agonism on aGVHD enables development of future therapeutic approaches for aGVHD treatment, such as myeloid APC suppression.
Supported by an Edward P. Evans Foundation Young Investigator Award, a Conquer Cancer Foundation American Society of Clinical Oncology Career Development Grant, and an American Cancer Society Clinician Scientist Development Grant
Dendritic cell-based amelioration of graft versus host disease in humanized mice
Authors: Kalinina Olga , Greene Anna , Knight Katherine L 
Published: January 2023

Abstract
Graft versus host disease (GVHD) is a severe, often lethal, complication of hematopoietic stem cell transplantation, and although prophylactic regimens are given as standard pre-transplantation therapy, up to 60% of these patients develop aGVHD, and require additional immunosuppressive intervention. Previously we showed that treatment of mice with a purified probiotic molecule, exopolysaccharide (EPS) from Bacillus subtilis, significantly increased survival of GVHD mice compared to control PBS-treated mice (70% vs 10%, respectively), with EPS-treated mice surviving more than 100 days after aGVHD induction. To test if EPS can also ameliorate GVHD in humans, we used humanized NSG-HLA-A2 mice and induced GVHD by i.v. injection of HLA-A2neg human PBMCs. After deriving dendritic cells (DC) from CD34+ cord blood cells, we pretreated them with EPS or PBS, and injected them together with PMBCs into the NSG-HLA-A2 mice. We found that EPS-treated DC significantly prolonged the survival of these mice, compared to PBS-treated DC. We also found that EPS-treated DC downregulated expression of activation markers (CD80, CD86), upregulated expression of inhibitory molecules (PD-L1, PD-L2), and inhibited activation of alloreactive T cells in MLR. We conclude that EPS induces tolerogenic human DC, which ameliorate GVHD in humanized NSG-HLA-A2 mice, likely by inhibiting alloreactive T cell response.
Supported by NIH R41 AI155281
IL-10 Signaling in T Cells is Essential for Transplant Tolerance Induction
Authors: Raimondi Giorgio , Lozano Marcos Igleasias , Komin Alexander , Chicco Maria 
Published: January 2023

Abstract
Costimulation blockade (CoB)-based immunotherapy is a promising approach for transplant recipients yet to be realized. Elucidating which factors impact the efficacy of CoB is a necessary step to enable clinical applicability. In this regard, pro- and anti-inflammatory cytokines are being recognized as having an impact on T cell activation beyond effector differentiation. We then aimed at elucidating the role that direct IL-10 signaling in T cells has on the outcome of CoB.
We employed a mouse skin transplt model were Balb/c skin was transplanted to wt C57BL/6 (B6) mice or to B6 with T cell-restricted expression of a dominant negative IL-10 receptor (10R-DN). Recipients received peri-transplant donor specific infusion and three weekly anti-CD154 mAb doses. Unmanipulated 10R-DN recipients rejected their transplant with dynamics identical to wt B6. However, differently from wt B6, graft survival in 10R-DN could not be promoted by CoB (MST 105d vs 30d in the latter) revealing a novel and important effect of IL-10 on T cells. This accelerated rejection correlated with increased production of TNF-α, IFN-γ and IL-17 by T cells in draining lymphoid tissues. MLR experiments indicated that despite IL-10 signaling is not involved in the ability of anti-CD154 to modulate alloreactive T cell proliferation, absence of this pathway increased the differentiation of TH1 effector cells. Moreover, the same experiments indicated that lack of IL-10 signaling in T cells impaired induction of Tregs.
Overall, our results reveal previously unappreciated roles of IL-10 signaling in T cells that represent potential “Achille’s Heels” in the successful implementation of CoB and suggest complementary interventions for the actuation of robust immunoregulation.
Supported by a grant from JDRF (2-SRA-2016-304-S-B)
Oral alloantigen exposure promotes donor-specific tolerance in a mouse model of minor mismatched skin transplantation
Authors: Wang Peter , Chen Luqiu , McIntosh Christine M , Lane Jorden I , Li Rena , Xie Stephen Z , Sattar Husain , Esterhazy Daria , Chong Anita S F , Alegre Maria-Luisa 
Published: January 2023

Abstract
Oral tolerance is a powerful, non-invasive route to induce immune tolerance, yet it has been unsuccessful to promote graft acceptance in rodent models of transplantation. To this end, we developed a mouse model where the model antigen ovalbumin (OVA) was introduced orally, in a regimen designed to improve bioavailability, duodenal lymph node drainage and prolong duration of antigen exposure, prior to transplantation with skin grafts constitutively expressing membrane-bound OVA. We found that oral OVA treatment pre-transplantation promoted permanent graft acceptance as well as linked tolerance to skin grafts expressing OVA coupled to the additional model antigen 2W. Tolerance was donor-specific, as secondary donor-matched, but not third-party grafts were spontaneously accepted in the absence of additional immunosuppression. Treatment with this oral OVA regimen promoted an anergic phenotype in OVA-reactive CD8+ T cells, generation of iTregs, and dysfunction of OVA-reactive CD4+ T cells pre-transplantation. The tolerance induced by oral OVA was robust, as anti-CD25/anti-PD-L1 at the time of transplantation failed to prevent skin graft acceptance. Mechanistically, we revealed a critical role for the proximal gut draining lymph nodes (gLNs) in mediating this effect, as an intestinal infection that drains to the proximal gLNs prevented the induction of transplantation tolerance. Our study extends previous work applying oral antigen exposure to transplantation and serves as proof of concept that the systemic immune mechanisms supporting oral tolerance are sufficient to promote long-term graft acceptance. Regimens that improve bioavailability and duration of oral alloantigen administration may hold promise in the clinic.
This work was supported by a NIAID grant to ASC and MLA (P01AI-97113). Peter Wang was supported by undergraduate research grants from the Katen Scholars Program and the Dean’s Fund for Undergraduate Research.
Graft-Matched Pregnancy Imparts Profound Epigenetic Changes onto Alloreactive Memory T Cells to Enforce a Phenotypic and Transcriptional State of Tolerance
Authors: Pollard Jared , Yin Dengping , Mandal Malay , Gounari Fotini , Alegre Maria-Luisa , Chong Anita 
Published: January 2023

Abstract
Pregnancy is an immunological paradox, where the maternal immune system develops tolerance to a semi-allogeneic fetus, yet pregnancy is a major sensitizing event in clinical transplantation. We previously reported that pregnancy robustly tolerizes fetus-specific T cells, allowing for the spontaneous acceptance of fetus-matched allografts when B cells and alloantibodies are absent at the time of transplantation(1). In this study, we report that mice sensitized to 2WOVA.B/c allogeneic skin grafts (Memory) underwent successful pregnancies comparable to Naïve mice when mated with 2WOVA.B/c males. We utilized a high-dimensional multi-omics approach to characterize post-partum (PP) fetus-specific T cells from Naïve or Memory mice after semi-allogeneic pregnancies. Strikingly, pregnancy induced fetus-specific naïve and memory T cells to achieve a nearly identical phenotype, characterized by upregulation of co-inhibitory molecules FR4, CD73, PD1, and CTLA4 in CD4+FoxP3- Tconvs and upregulated PD-1, FR4 and Lag3 in CD8+ T cells. We confirmed that naïve and memory PP CD8+ T cells acquired similar transcriptional profiles via RNA-Seq; unexpectedly, pregnancy induced profound epigenetic modifications in memory, but not naïve, CD8+ T cells. In vitro studies indicated reduced TNF aproduction by memory PP CD4+ and CD8+ T cells, and experiments examining their response to fetus-matched allografts are ongoing. Our studies suggest that significant epigenetic imprinting is necessary to constrain memory T cells to preserve fetal viability during semi-allogeneic pregnancy. These findings may provide insights into controlling antigen-specific memory T cells, a major barrier to clinical transplant success
Evaluation of checkpoint inhibitor therapies using a mixed lymphocyte reaction (MLR) assay
Authors: Trigg Jasmine E , McBain Kirsty , Liu Zhaoping , Dale Timothy , Szybut Clare 
Published: January 2023

Abstract
Checkpoint inhibitors block the ‘off’ signals that are engaged when T cells interact with antigen-presenting cells, such as dendritic cells (DCs). This increases T cell activation and enhances the anti-tumor response. Here we demonstrate the use of advanced flow cytometry to evaluate these immunotherapies in vitro by co-culturing immune cells from two donors in an MLR assay.
Assays were set up as either one-way or two-way MLR co-cultures. One-way assays combined isolated DCs from one donor with CD4+ T cells from another. Two-way assays mixed PBMCs from two donors. Cells and checkpoint inhibitors were plated in 96 or 384-well formats for 3–6 days. Samples taken during the time course were analyzed for cell subsets and cytokine concentrations using the iQue® Advanced Flow Cytometry Platform.
Expression of activation marker CD25 on T cells was 3-fold greater after 6 days in co-culture with DCs (62 ± 2 %) compared to T cells in monoculture (22 ± 9 %). The addition of an anti-PD1 checkpoint inhibitor induced a further, concentration dependent increase in CD25 expression. This was accompanied by increased production of cytokines indicative of activation and inflammation, such as TNFα and CCL2. Throughout the study, there were large differences in sensitivity to drug response between donor pairs. For example, IFNγ release increased 13.5-fold upon addition of anti-CTLA4 (17 ng/mL) with one donor pair, compared to only 1.3-fold with another. This highlights the need to characterize drug activity using multiple immune cell donor pairs.
These data exemplify the use of advanced flow cytometry to generate pharmacological data for checkpoint inhibitor activity on T cells in MLR, with the potential to profile libraries of novel therapeutics in minimal time.
Sialylation of CD44 by ST8sia6 is required for recognition by the inhibitory receptor Siglec-7
Authors: Chen Yin , Friedman David J , Saraswat Mayank , Shapiro Michael J , Wilfahrt Drew , Pandey Akhilesh , Shapiro Virginia Smith 
Published: January 2023

Abstract
Glycans decorate almost all proteins and lipids on the cell surface, and they play a critical role in cell-cell interactions. Terminal sialic acid addition to glycans helps the immune system distinguish between “self” and “non-self.” The enzyme ST8sia6 adds alpha-2,8 disialic acids to cell surface receptors, and this sialic acid addition promotes binding to the inhibitory receptor Siglec-E. We have previously shown ST8sia6 overexpression in tumors accelerates tumor growth in a Siglec-E dependent manner. Proteomic analysis in murine tumor lines identified CD44 as a target for ST8sia6. CD44 expression on tumors is associated with metastasis and chemoresistance. Since humans and mice have different sialic acid structures, we examined whether ST8sia6 makes CD44 a ligand to the ortholog of Siglec-E in human cells, Siglec-7. When HEK293T cells overexpress ST8sia6 and CD44 together, Siglec-7 binding was significantly increased compared to ST8sia6 or CD44 overexpression alone. Thus, ST8sia6 has a conserved role from mouse to human in decorating CD44 with sialic acid. Our study proposes that ST8sia6 sialylation of CD44 could suppress immune responses and enables cancer progression in human.
Supported by grant: 2T32AI007425-23 to DJF and R01 CA243545-01S1 to V.S.S.
Organoid models for high-grade endometrial cancer to dissect tumor immune microenvironments
Authors: Chung Charlie , Nizam Aaron , Goldberg Gary L , Beyaz Semir 
Published: January 2023

Abstract
Inactivation of antigen processing and presentation (APP) pathways is one of the major mechanisms for immune evasion in many cancer types including high-grade endometrial cancer (HGEC). Impairments in APP pathways may contribute to the low immunotherapy response rate among the patients. However, lack of pre-clinical models for HGEC poses a significant challenge in studying critical therapeutic aspects of the cancer such as cancer-immune cell interactions. To our knowledge, we have established the largest endometrial biobank comprising primary tissues, patient-derived organoids (PDO), and their matching immune cells from all existing endometrial cancer types, enabling us to establish the interaction between cancer and immune cells in vitro.
Using a subset of our biobank, we performed RNA-seq on 68 PDOs and found that genes involved in APP pathways are downregulated in HGEC organoids compared to their matching normal pairs. We found that MHC-I and -II could be upregulated by IFNg and Tazemetostat, an EZH2 inhibitor. We further assessed the effects of enhanced MHC-I and -II expression mediated by the two molecules on cancer-immune cell interactions by setting up syngeneic organoid-immune cell co-cultures. We found that HGEC organoids that were pre-treated with IFNg or Tazemetostat displayed a greater immune cell co-localization and immune cell-mediated apoptosis during the co-culture period.
Our proof-of-principle experiments demonstrate the utility of our co-culture system in studying the cancer-immune cell interactions in vitro. Our system could be used to test the efficacy of current therapeutics or to screen potential drugs which enhance immune responses against HGEC, which lacks effective treatment options.
This research was supported by grants from The Mark Foundation for Cancer Research (20-028-EDV630), NCI CancerCenter Support Grant (5P30CA045508),and CSHL-northwell transitional research fund.
Turning immunologically cold tumors into hot ones by activating hepatoma-intrinsic FADD/NF-κB/CCL5 pathway
Authors: Lu Jiahuan , Chan Anthony Wing Hung , To Ka Fai , Zhou Jingying 
Published: January 2023

Abstract
Lymphoepithelioma-like hepatocellular carcinoma (LEL-HCC) is a distinct variant of HCC that displayed immunologically hot tumor features with prominent tumor-infiltrating CD8+ T lymphocytes (TILs). Our whole exosome sequencing data pointed out an increased prevalence of chromosome 11q13.3 amplification in LEL-HCC, in which 13/17 genes located in 11q13.3 were upregulated. Among these genes, fas-associated death domain (FADD) displayed a strongest correlation with differentially upregulated immune regulatory genes in HCC with 11q13.3 amplicon. In addition, CD8+ T cells preferred to migrate into tumors with higher FADD expression in vitro and in vivo. Mechanistically, FADD activated nuclear factor-κB (NF-κB) that in turn promoted T cell trafficking chemokine C-C motif chemokine ligand 5 (CCL5) production. Furthermore, Fadd expression was positively correlated with Ccl5 and TIL proportions, leading to increased immune checkpoint blockade (ICB) responsiveness in orthotopic HCC mouse models. As LEL-HCC patients also exhibited a concordant higher expression of FADD and CD8, our findings suggested that activation of FADD may turn cold tumors into hot ones and improve HCC ICB responsiveness.
This project is supported by grants from Health and Medical Research Fund (07180556) and General Research Fund 2019/20 (14108219).
Ovarian cancer cell glucocorticoid receptor activity modulates cytokine secretion promoting infiltration of immunosuppressive cells into the tumor microenvironment
Authors: Taya Manisha , Gonzalez Janet , Bennett Lynda , Conzen Suzanne D 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) have been identified as a heterogeneous cell population that expand during tumor progression and infiltrate the tumor, peripheral blood, as well as ascites of ovarian cancer patients. MDSCs contribute to tumor immune tolerance via inhibition of T-cell proliferation/activation. It is also known that high tumor cell glucocorticoid receptor (GR) expression is associated with a relatively poor prognosis in ovarian cancer patients. We hypothesized that ovarian tumor cell GR expression and activity may modulate tumor cell cytokine secretion thereby leading to increased MDSC generation and recruitment, thus creating an immunosuppressive tumor microenvironment. Our preliminary data confirms GR-mediated secretion of pro-tumorigenic cytokines including G-CSF, TGF-b, and MCP-1 in high-grade ovarian cancer models. We also observed GR-mediated downregulation of IL-2 and IFN-g, cytokines required for anti-tumor T-cell function. Importantly, treatment of cells with a competitive GR antagonist reversed this immunosuppressive secretome effect. Moreover, culturing healthy peripheral blood mononuclear cells (PBMCs) with tumor conditioned media from GR activated ovarian cancer cells demonstrated that secreted immunomodulatory factors have the capacity to promote differentiation of human MDSCs from PBMCs. Based on the insight provided by these data, we propose to determine whether ovarian tumor cell-intrinsic GR activation and resulting MDSC generation has an inhibitory effect on cytotoxic T-cell function, contributing to earlier relapse of GR-positive ovarian cancers, and eventually whether this mechanism may be targeted to improve patient outcomes in this subset of ovarian cancers.
Supported by grant from CPRIT
Transcriptome analysis identifies networks and key drivers in tumor microenvironment for metastatic castration-resistant prostate cancer
Authors: McKinney Lawrence P , Singh Rajesh , Jordan I King , Varambally Sooryanarayana , Dammer Eric B , Sanda Martin G , Lillard James W 
Published: January 2023

Abstract
Prostate cancer (PCa) is the second most cancer-related cause of death in men. The most lethal form of PCa is metastatic castration-resistant prostate cancer (CRPC) that has progressed to the bone. The molecular mechanism underlying PCa progression to bone has not been fully elucidated. Here, we identify gene networks and characterize the cell types that contribute to the heterogeneity and complexity of CRPC associated with bone metastasis. Networks were investigated by using a novel weighted gene network co-expression analysis (WGCNA) method and examining overrepresentation of upstream regulators and signaling pathways within co-expressed transcriptome modules across clinically annotated transcriptomes from mCRPC patients (N=60). Functional Enrichment analysis was used, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein-protein interaction analysis to identify biological functions of related hub genes overrepresented in our module of interest. WGCNA identified significant modules that are associated with CRPC bone metastasis. KEGG and GO analysis results revealed genes in these modules were mainly related to collagen formation, cell signaling peptides, and bone regulation processes. Genes positively correlated with bone metastasis exhibited the following biological pathways: PI3K-Akt signaling, ECM-receptor interaction, and protein digestion and absorption pathways. This study provides novel insights into the biological pathways associated with CRPC metastasis to the bone and its tumor immune microenvironment (TIME) niche. The modules associated with bone metastasis and overall survival represent both known and novel pathways.
Supported by NIH/NCI Grant: MSM/Tuskegee U/UAB Comp. Cancer Center Partnership Name & Number of Grant Award: NCI - U54CA118638