bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transcriptome analysis of circulating lymphoid progenitors after allogeneic stem cell transplantation in humans reveals profound alterations of cellular metabolism (TRAN1P.930)
Authors: Toubert Antoine , Glauzy Salome , André-Schmutz Isabelle , Lachuer Joel , Peffault de Latour Regis , Servais Sophie , Socié Gerard , Clave Emmanuel 
Published: January 2023

Abstract
T-cell reconstitution is delayed by acute Graft-Versus-Host Disease (aGVHD) during allogeneic hematopoietic stem cell transplantation (allo-HSCT). Here, we analyzed transcriptional programs of cell-sorted circulating lymphoid progenitors and CD34+Lin-CD10- non lymphoid progenitors in 12 allo-HSCT patients having (n=4) or not developed (n=8) grade 2 aGVHD and in 4 age-matched healthy donors. Gene expression was quantified on the GeneChip HG u133 plus 2.0. Pathway analyses were performed with KEGG, GSEA and DAVID. Major deregulated pathways included protein synthesis, energy production, cell cycle regulation and response to stress. Notably, Genes from ribosome protein biogenesis, translation machinery (EEF1D, EEF1G, EIF3K) and cell cycle (CCND1, CDK6) were over-expressed in progenitors from patients without aGVHD compared with those from patients affected by aGVHD and from healthy donors. Expressions of genes from the oxidative phosphorylation metabolic pathway (NDUFS2, SDHA, ATP5A1) and genes involved in stress resistance (BTG2, MGST3, HPX) were specifically increased in CD34+Lin-CD10+CD24- lymphoid progenitors from patients without aGVHD. In all, we show that circulating lymphoid T-cell progenitors undergo profound changes in metabolism favoring energy production and response to stress after allo-HSCT in humans. These mechanisms are abolished in case of aGVHD, indicating a persistent cell-intrinsic defect in addition to the impact of aGVHD on the bone marrow environment.
Donor CD103+ dendritic cells in the colon engender lethal graft-versus-host disease (TRAN1P.938)
Authors: Koyama Motoko , Cheong Melody , Markey Kate , Gartlan Kate , Kuns Rachel , Locke Kelly , Lineburg Katie , Teal Bianca , Bunting Mark , Vuckovic Slavica , Zhang Ping , Teng Michele , Varelias Antiopi , Tey Siok-Keen , Wockner Leesa , Engwerda Christian , Smyth Mark , Belz Gabrielle , McColl Shaun , MacDonald Kelli , Hill Geoffrey 
Published: January 2023

Abstract
The spatial and temporal nature of donor antigen presentation during graft-versus-host disease (GVHD) and its relevance to disease severity are poorly understood. Here we use luciferase expressing TCR transgenic CD4+ T cells specific for indirect alloantigen presentation within MHC class II to demonstrate that GVHD markedly enhances indirect presentation within the mesenteric lymph nodes (mLN). By using antibody specific for alloantigen complexed with donor MHC and IRF4 and CCR7 deficient donors we demonstrate that this presentation is entirely mediated by the donor CD103+CD11bneg dendritic cell (DC) subset that migrates from the colon under the influence of CCR7. We used IL-12, IL-6, MyD88/TRIF and receptor for advanced glycation end products (RAGE) deficient grafts to demonstrate that this differentiation of donor T cells in the mLN during GVHD is driven by high levels of alloantigen and local IL-12 and IL-6 secretion elicited by Toll-like receptor and RAGE signals. Notably, alloantigen presentation by CD103+ DC migrating from the colon imprints gut-homing integrin signatures on donor T cells in the mLN, leading to their emigration into the gastrointestinal (GI) tract where they mediate fulminant GVHD. These data highlight multiple therapeutic targets and the ability of GVHD, once initiated by recipient APC, to generate a profound and localized feed-forward cascade of indirect antigen presentation and cytokine secretion within the GI tract, culminating in lethality.
Jak/STAT signaling inhibition enhances costimulation blockade and promotes transplant acceptance (TRAN1P.943)
Authors: Raimondi Giorgio , Khalifian Saami , Miller Devin , Oh Byoungchol , Lee WP , Brandacher Gerald 
Published: January 2023

Abstract
Transplant tolerance induction through costimulation blockade (CoB) remains an elusive goal. Recent evidence suggests that inflammatory cytokines (IC) contribute to the activation of alloreactive T cells in a CD28- and CD40-independent manner. We aimed to delineate the possible synergism between CD28 blockade and inhibition of IC signaling via Jak/STAT inhibition. First, we measured the impact on mouse T cell activation via quantification of IL-2 secreting cells at 24h post in vitro stimulation. Although CTLA4-Ig diminished the proportion of IL-2+ cells, the concomitant addition of the supernatant (MATSup) from maturing dendritic cells completely abolished this effect. This suggested an early impact of IC on T cell activation. MATSup counteracted the anti-proliferative effect of CTLA4-Ig on both CD4 and CD8 T cells. However, addition of the Jak3/1 inhibitor Tofacitinib (Tofa) completely restored the effect of CTLA4-Ig in presence of MATSup. In a BALB/c to B6 heart transplant model a short course of Tofa (d0-6) synergized with CTLA4-Ig promoting long term graft survival (MST untreated: 11d, CTLA4-Ig only: 36d, Tofa+CTLA4: 114d). Survival was associated to lower Th1 cell production and, unexpectedly, an increment in graft infiltrating Treg. Overall, our results indicate that inflammatory cytokines counteracts the efficacy of CoB. However, this effect can be neutralized via transient inhibition of Jak signaling - a promising new immunoregulatory strategy we define Enhanced CoB.
Human mesenchymal stem cell-educated macrophages: a potential therapy for GVHD and radiation injury (TRAN1P.944)
Authors: Bouchlaka Myriam , Kim Jaehyup , Bloom Debra , Hematti Peiman , Capitini Christian 
Published: January 2023

Abstract
Total body irradiation is typically used to condition patients for allogeneic hematopoietic stem cell transplant, leading to release of inflammatory cytokines from damaged tissues. After transplant, many patients develop graft-versus-host disease (GVHD) resulting from donor T cells attacking host tissues leading to further inflammation. Mesenchymal stem cells (MSCs) have immunosuppressive and tissue repair properties, but clinical trials using MSCs to treat GVHD have shown mixed results. Macrophages (MQs) are important regulators of immunity and can promote tissue remodeling. We have previously shown that MSCs can educate MQs toward a unique anti-inflammatory phenotype, however the implications for in vivo models of inflammation have not been studied. Using flow cytometry, we found that in comparison to MQs, mesenchymal stem cell-educated macrophages (MEMs) have increased expression of the inhibitory molecules PD-L1 (p=0.039), PD-L2 (p=0.0002), and markers of alternatively activated macrophages: CD206 (p=0.04) and CD163 (p= 0.0004). MEMs also show increased expression of TGF-β (p&lt;0.0001), Arginase-1 (p&lt;0.0001) and IL-6 (p=0.0006), and decreased expression of IL-12 (p=0.004) and TNF-α (p&lt;0.0001) by RT-PCR. In two xenogeneic mouse models, we show that MEMs significantly enhance survival from lethal GVHD (p=0.028 vs MSC) and from lethal radiation (p=0.0044 vs MSC). MEMs could be a novel cellular therapy for the management of GVHD and protection from radiation-induced injury.
Delayed CTLA-4Ig treatment of established acute allograft rejection successfully abrogates ongoing B cell responses (TRAN1P.950)
Authors: Young James , Chen Jianjun , Yin Dengping , Sciammas Roger , Chong Anita 
Published: January 2023

Abstract
Once rejection is initiated, halting the immune response becomes much more difficult and effective therapies are necessary. Despite evidence that CTLA4Ig is ineffective at controlling memory T cell responses, we here report that CTLA4Ig is unexpectedly effective at controlling established B cell responses and treating established acute rejection. When C57BL/6 mice were sensitized with BALB/c splenocytes and treated with CTLA4Ig from 7d post-sensitization, B cell germinal center responses rapidly collapsed and alloantibody production was halted. In contrast, the effector CD4 and CD8 T cells response, which was monitored with seeded TCR-tg or endogenous allospecific cells, was not inhibited, and the extent of CD44 differentiation, IFNg production, and Tfh differentiation was similar to untreated controls. We also tested the effect of delayed CTLA-4Ig in established acute allograft rejection. In C57BL/6 recipients of BALB/c hearts, delayed treatment of CTLA4Ig till 6d post-transplant (just 2d before rejection) inhibited alloAb production and prevented acute rejection. 10 of 15 grafts survived to ≥ 30d post-transplant, despite significant T cell infiltration in the graft. Thus significant control of B cell responses and rejection can be achieved through delayed CTLA-4Ig treatment. This study raises the exciting possibility of using CLTA-4Ig therapeutically, not just prophylactically, in the clinic to control established B cell responses and established acute rejection.
The adaptor protein ASC controls transplantation outcomes independently of the inflammasome (TRAN1P.951)
Authors: Cheong Melody , Gartlan Kate , Tey Siok-Keen , Kuns Rachel , Lor Mary , Lineburg Katie , Teal Bianca , Shi Wei , Raju Jyothy , Zhang Ping , Varelias Antiopi , Leveque-El Mouttie Lucie , Olver Stuart , Bunting Mark , Lane Steven , Boyle Glen , Ting Jenny , Schroder Kate , Engwerda Christian , Khanna Kum Kum , Smyth Mark , MacDonald Kelli , Koyama Motoko , Hill Geoffrey 
Published: January 2023

Abstract
The adaptor protein ASC is known to facilitate caspase-1 activation essential for innate host immunity via the formation of the inflammasome complex - a multi-protein structure responsible for processing IL-1b and IL-18 to their active moieties. Here we report for the first time, a unique inflammasome-independent role for ASC in the control of transplant outcome following allogeneic bone marrow transplantation (BMT). We demonstrate that ASC-deficient donor CD8+ T cells fail to induce GVHD lethality due to an inability to differentiate into fully cytolytic effectors after BMT, with a developmental bias instead towards CD127+KLRG1- memory CD8+ T cells. Despite this, graft-versus-leukaemia effects against BCR-ABL NUP98/HOXA9 primary leukemia remained largely intact. This phenomenon was inflammasome independent, since GVHD lethality and T cell differentiation were not altered in recipients of caspase-1-deficient T cells. We also confirmed a reduced capacity for human T cells in which ASC was knocked down by shRNA to induce xenogeneic GVHD. In a model of bone marrow rejection, ASC expression in recipient CD8+ T cells profoundly impaired graft rejection and was permissive of robust engraftment across MHC barriers and long term survival. Taken together, these findings demonstrate an inflammasome-independent role for ASC in controlling GVHD and graft rejection. Thus, the inhibition of ASC in the clinic represents an important new therapeutic target to manipulate transplant outcomes.
Hyperglycemia is indispensable for delayed islet graft rejection in STZ-induced diabetes mouse model. (TRAN1P.952)
Authors: Bogacz Marek , Drutel Robert , Leanhart Silvia , Kern Joanna , Pacholczyk Rafal 
Published: January 2023

Abstract
Transplantation of allogeneic pancreatic islets or pancreas is one of the methods to control euglycemia in patients with type 1 diabetes. However despite emergence of new tolerance induction protocols these patients require lifelong immunosuppression which is associated with many adverse effects including toxicity towards transplanted pancreatic β-cells. It has been previously shown that streptozotocin (STZ)-induced diabetes can delay or abrogate rejection of allografts in CBA/J or C57BL/6 mice. To test this effect on autoimmune prone background we performed a series of allogeneic-islet transplantation experiments using NOD mice as recipients. Our results show that if transplanted during specific time window after STZ treatment (2-3 days) graft rejection will be delayed but not completely abrogated. Using insulin pump to control blood glucose level we demonstrate that STZ administration without episode of acute hyperglycemia is not sufficient to delay graft rejection. It is despite the fact that STZ injection alone results in increased frequency of regulatory CD4Foxp3+ T cells. Comparison of gene expression in CD4 T cells between STZ-treated and untreated mice shows selective increase of apoptosis in effector but not regulatory T cells.
Generation, cryopreservation, function and in vivo persistence of ex vivo expanded cynomolgus monkey Treg (TRAN1P.927)
Authors: Guo Hao , Zhang Hong , Lu Lien , Ezzelarab Mohamed , Thomson Angus 
Published: January 2023

Abstract
Foxp3+ regulatory T cells (Treg) function to promote immune tolerance and prevent immunopathogenesis. Preclinical studies have demonstrated that adoptive transfer of Treg prevents rejection of organ allografts in mice and humanized mouse models. In this study, we optimized a protocol to expand flow-sorted cynomolgus Treg &gt;1000-fold after three rounds of stimulation with anti-CD3 mAb-pulsed artificial APCs, rapamycin and IL-2. The expanded Treg from each round maintained high level expression of Treg signature markers, -CD25, CD27, CD39, Foxp3, Helios, CTLA-4, as well as CXCR3, a homing receptor of Th1-like cells. In contrast to expanded Teff, Treg produced minimal IFN-γ and IL-17 and no IL-2. In addition, the expanded Treg from each round showed high suppressive capacity in vitro. Compared to freshly-expanded Treg, Treg immediately thawed from cryopreserved stocks maintained Treg signature markers and suppressive capacity, but were less viable. However, additional rounds of stimulation/expansion restored maximal viability. Adoptively-transferred autologous Treg labeled with CFSE (approximately 2x107/kg, i.v.) could be detected in vivo 2-3 months post infusion in peripheral blood, inguinal LN, mesenteric LN and spleen. Moreover, these cells maintained CD25, CCR7, CXCR3 expression and acquired CD62L after infusion in vivo.
c-Met identifies a population of matrix metalloproteinase 9-producing monocytes in peritumoral stroma of hepatocellular carcinoma (TUM6P.966)
Authors: Wu Yan , Zhao Lan , Zheng Limin 
Published: January 2023

Abstract
Macrophages (Mφ) are prominent components of solid tumors and exhibit distinct phenotypes in different microenvironments. Previously, we found that tumors could alter the normal developmental process of Mφ to trigger transient activation of monocytes in the peritumoral stroma of human hepatocellular carcinoma (HCC). In the present study, we showed that a fraction of monocytes in the peritumoral stroma, but not in HCC cancer nests, expressed surface c-Met molecules. Monocytes exposed to tumors strongly expressed c-Met proteins with kinetics similar to their activation status, and significant correlations were found between c-Met levels and HLA-DR expression on tumor-infiltrating monocytes. NF-κB-mediated autocrine TNF-α stimulated the expression of c-Met on activated monocytes, and by interacting with its ligand HGF, c-Met regulated the retention, accumulation, and matrix metalloproteinase (MMP) 9-producing capacity of monocytes in the peritumoral stroma of HCC. The intensity of c-Met expression on tumor-infiltrating monocytes was associated with high mortality and reduced survival of patients with HCC. Therefore, the expression of c-Met on activated monocytes/Mφ may represent a novel mechanism by which a tumor actively and precisely regulates the distribution and functions of these cells to facilitate disease progression.
Suppression of tumor growth by a M2b macrophage reverter in humanized murine chimeras created with peripheral blood monocytes of patients with advanced stage hepatocellular carcinoma (TUM6P.967)
Authors: Tsuchimoto Yusuke , Asai Akira , Ohama Hideko , Yokohama Keisuke , Nakamura Ken , Fukunishi Shinya , Tsuda Yasuhiro , Higuchi Kazuhide , Kobayashi Makiko , Suzuki Fujio 
Published: January 2023

Abstract
Previously, we have reported that humanized murine chimeras created with peripheral blood monocytes from patients with early stage hepatocellular carcinoma (E-hccM) are resistant against tumor inoculation, while tumors grow in the same chimeras created with advanced stage patient monocytes (A-hccM). A majority of E-hccM are shown to be M1 monocytes, and a predominance of M2b monocytes is shown in A-hccM. In this study, we tried to suppress tumor growth in humanized chimeras created with A-hccM by CCL1 antisense ODN treatment, which reverts Mf from M2b to resident. Chimeras, created in X-irradiated NSG mice after inoculation of A-hccM (i.v., 1 x 106 cells/mouse, every 2 weeks), were inoculated with HepG2 cells (2 x 106 cells/mouse, s.c.). CCL1 antisense ODN (6 mg/mouse) were administered 4 times within 36 hrs of each A-hccM inoculation. In the results, 843 ± 227 mm3 of tumors were detected in the chimeras inoculated with A-hccM 8 weeks after the tumor inoculation, while tumors were not demonstrated in A-hccM chimeras treated with CCL1 antisense ODN, and 753 ± 165 mm3 of the tumors were detected in A-hccM chimeras treated with scrambled ODN. A-hccM were characterized as IL-12-IL-10+CCL1+iNOS-CD14+ cells, while IL-12+IL-10-CCL1-iNOS+CD14+ cells with tumoricidal activities were detected in A-hccM treated with the antisense ODN. These results indicate that M2b monocytes present in A-hccM play a key role in the tumor growth seen in advanced stage hepatocellular carcinoma patients.
Emodin inhibits breast cancer growth by modulating the phenotype of tumor-associated macrophages (TUM6P.1004)
Authors: Iwanowycz Stephen , Wang Junfeng , Jia Xuemei , Fan Daping 
Published: January 2023

Abstract
Infiltration of macrophages into breast tumors is correlated with increased metastasis and decreased patient survival. Activated macrophages display a spectrum of phenotypes, at either end of which are pro-inflammatory/anti-tumor M1 or anti-inflammatory/pro-tumor M2 macrophages. Tumors recruit macrophages to the primary site as well as to pre-metastatic niches, educate them to adopt an M2-like phenotype, and thereby create an immunosuppressive microenvironment. We previously showed that Chinese herb derived compound emodin has the potential to inhibit tumor metastasis by targeting tumor cell-macrophage interactions. Here we tested the ability of emodin to inhibit breast cancer growth in mice and examined the underlying mechanisms. Emodin was used to treat mice bearing EO771 or 4T1 breast tumors. We found that emodin treatment reduced M2-like macrophages in the primary tumors and the lungs, blunted the growth of primary tumors, and attenuated lung metastasis. Further in vitro studies showed that emodin modulates macrophage polarization and function through affecting multiple intracellular signaling pathways as well as through possible regulation of the epigenetic landscape. These results suggest that emodin can be developed as an anti-breast cancer therapy by its virtue of modulating the phenotype of tumor-associated macrophages.
Targeting dysfunctional myeloid cells delays disease development and improves immune function in a CLL mouse model (TUM6P.1009)
Authors: Hanna Bola , McClanahan Fabienne , Zaborsky Nadja , Dürr Claudia , Kalter Verena , Egle Alexander , Lichter Peter , Gribben John , Seiffert Martina 
Published: January 2023

Abstract
Previous in vitro studies have shown that myeloid cells play a key role in apoptosis-resistance in chronic lymphocytic leukemia (CLL). However, their composition and function have not been thoroughly investigated in vivo. Using the Eµ-TCL1 mouse model of CLL, we observed severe skewing of myeloid cell populations along with CLL development. Monocytes and M2-like macrophages infiltrated the peritoneal cavity of leukemic mice. Monocytes expressing high levels of inflammatory factors like IL-10, TNFα and CXCL9 accumulated also in the spleen, the main site of disease, and were severely skewed towards Ly6Clow phenotype. Along with that, the percentage of MHC-IIhi dendritic cells (DCs) and macrophages significantly dropped in the spleen, suggesting monocyte differentiation arrest. Myeloid cell depletion using liposomal clodronate resulted in a significant control of CLL development, repair of innate immune cell phenotypes and partial resolution of systemic inflammation. Also, CLL-induced loss of naive T cells was resolved. CLL-associated monocytes and DCs aberrantly express high PD-L1 levels suggesting their contribution in T cell defects. Importantly, PD-L1 blockade very effectively prevented CLL development and was accompanied with a re-activation of immune effector cells. This included restoration of mature macrophages and MHC-IIhi DCs. Our preclinical data suggests that targeting myeloid cell survival and immunosuppression can serve as a novel strategy for CLL immunotherapy.
Induction of cell death following restimulation of the T cell receptor results in activation of the DNA damage response pathway in a JNK and ROS-dependent manner (TUM7P.1026)
Authors: Scheffel Matthew , Scurti Gina , Simms Pat , Garrett-Mayer Elizabeth , Nishimura Michael , Mehrotra Shikhar , Voelkel-Johnson Christina 
Published: January 2023

Abstract
The p53 protein is activated by various forms of cellular stress and its function is modulated though a multiplicity of posttranslational modifications to dictate signaling outcomes. A recent study demonstrated that inhibition of p53 by pifithrin-m effectively blocked cell death caused by restimulation of the T cell receptor (TCR). Using human T cells, we investigated the phosphorylation status of p53 following TCR restimulation and detected a time-dependent increase in phosphorylation on Ser15, which correlated with onset of cell death. Phospho-p53 (Ser15) was limited to the nucleus and coincided with phosphorylation of ATM. Blocking ATM function through KU55933 or caffeine prevented cell death. Phosphorylation of ATM suggested activation of the DNA damage response pathway and we confirmed coordinate increases in both yH2AX and SMC-1. Activation of the ATM-p53-dependent DNA damage response pathway following restimulation of the TCR was reduced by inhibition of JNK by SP600125 as well as the anti-oxidant N-acetyl cysteine (NAC). Activation of the DNA damage pathway occurred in an antigen-specific manner and was also observed in T cells derived from melanoma patients undergoing adoptive T cell therapy. In vitro treatment with NAC improved survival of T cells while increasing melanoma cell killing. Thus our data highlight a role for activation of the DNA damage response pathway following TCR restimulation and its potential translational significance in adoptive T cell therapy.
PD-1+CD28H+ selectively identifies for superior tumor-reactive T lymphocytes in pancreatic cancer (TUM7P.1013)
Authors: Zhu Yuwen , Koenig Michelle , Byers Joshua , Paniccia Alessandro , schulick alex , schulick richard , Chen Lieping , edil barish 
Published: January 2023

Abstract
Tumor-infiltrating lymphocytes (TILs) are enriched in tumor-specific T cells and have been used as a source for adaptive cell therapy (ACT) of cancer. TILs are a mixture of different T cell subsets with both anti-tumor and pro-tumor capacities. It is valuable to isolate potent tumor-reactive T cells while eliminating suppressor T cells from TILs to maximize anti-tumor immune response. CD28H is a newly discovered co-receptor of the B7 family that interacts with its ligand B7-H5 to costimulate human T cells. CD28H is constitutively expressed on native T cells. However, repetitive antigenic exposure induces the loss of CD28H expression in T cells. T cells that have lost CD28H have the phenotypic characteristics of terminally differentiated and replicative senescent cells. Here we characterized PBMCs and TILs from patients with pancreatic cancer based on the expression of CD28H and PD-1. We found that a subset of T cells, PD-1+CD28H+ (DP) cells, are exclusively enriched in TILs. Based on phenotypic and functional analysis, our preliminary results suggest that PD-1+CD28H+ cells represent a subset of superior tumor-reactive cells in TILs. By targeting these DP cells by CD28H agonistic mAb and/or with a PD-1 blocking antibody, we are expecting to reprogram TIL functions in human pancreatic cancer.