bluuebunny/crossref_metadata_2025_split huggingface dataset:
DISTINCT IL-6 SIGNALING PATHWAYS DRIVE ALTERNATE PATHOGENIC T CELL DIFFERENTIATION AND GVHD AFTER TRANSPLANT IN VIVO
Authors: Wilkinson Andrew N. , Kuns Rachel D. , Varelias Antiopi , Vuckovic Slavica , Rose-John Stefan , Gartlan Kate H. , Hill Geoffrey R. 
Published: January 2023

Abstract

Introduction
Dysregulation of interleukin 6 (IL-6) is pivotal for the development of graft-versus-host-disease (GVHD) after allogeneic stem cell transplantation. IL-6 drives many protective as well as inflammatory effects, the latter involving differentiation of T cells into pathogenic Th22, Th17, and Tc17 subsets. IL-6 signals through three principal pathways; classical, trans, and cluster, but their contribution to T cell differentiation and GVHD remains unclear. Importantly, these pathways are differentially targeted by clinical IL-6 inhibitors.

Aims
To dissect the contribution of the IL-6 signaling pathways to T cell differentiation and GVHD in order to guide therapy.

Methods
Donor bone marrow and T cells were transplanted from lineage-specific Cre × IL-6fl/flor IL-6Rfl/fl mice into irradiated, MHC disparate, wild-type or sgp130:Fc transgenic recipient mice with or without IL-6R mAb. Donor T cell differentiation and GVHD was assessed after transplant.

Results
Surprisingly, IL-6 trans-signaling was not required for T cell differentiation and GVHD, but instead regulated Th22 cell development. Indeed, inhibition of IL-6 trans-signaling in sgp130:Fc recipients promoted the expansion of Th22 cells and generated severe cutaneous GVHD. In contrast, IL-6 classical-signaling was critical for the development of Th22 and Th17 cells and its inhibition significantly attenuated GVHD broadly. Interestingly, unlike Th17 differentiation, Tc17 differentiation was highly IL-6-dependent but independent of classical and trans-signaling pathways, consistent with induction by cluster-signaling.

Conclusion
These data highlight cluster, in addition to classical IL-6-signalling as key therapeutic targets.
Depletion of macrophages from Cytomegalovirus (CMV) latently infected donor allografts decreases CMV mediated immunopathology and prevents CMV-accelerated chronic rejection
Authors: Haese Nicole , Burg Jennifer , Ando Takeshi , Jones Iris , Kreklywich Craig , Orloff Susan , Streblow Daniel 
Published: January 2023

Abstract
Cytomegalovirus (CMV), a ubiquitous β-herpesvirus, is linked to acceleration of solid organ transplant vascular sclerosis (TVS) and chronic rejection (CR). Latent CMV infection increases cardiac-resident macrophages and T cells that may increase inflammation and promote allograft rejection. To investigate the role of cardiac macrophages in CMV mediated TVS/CR, macrophages were depleted from CMV+ donor allografts prior to transplantation using clodronate liposomes (clod) in our rat heart transplant model. Latently rat CMV infected donor F344 rats were treated with clod or PBS or control liposomes 3 days prior to heterotopic cardiac transplant into CMV-naïve Lewis rats and macrophage depletion was validated. Donor hearts were transplanted and monitored daily for rejection. Clod treatment significantly increased graft survival from 61 days post-transplant to 84 days. The severity of graft vessel disease was decreased compared to controls. Clod treatment significant decreased macrophage and T cell infiltration into allograft tissue versus controls. Clod treatment also altered allograft tissue protein and gene expression cytokine/chemokine profiles toward a phenotype that promotes graft survival by reducing the allograft inflammation. Clod treatment did not prevent CMV reactivation since all recipients seroconverted and CMV DNA was detectable in recipient salivary glands. Depletion of donor macrophages prior to transplant represents a novel method to modulate allograft rejection immunity and prevent the negative impact of CMV on allograft recipient survival.
PD-L1 expression of monocytes may be used a new marker of the classification for the cancer progression in hepatocellular carcinoma.
Authors: Asai Akira , Yasuoka Hidetaka , Tsuchimoto Yusuke , Ohama Hideko , Fukunishi Shinya , Tsuda Yasuhiro , Kobayashi Makiko , Higuchi Kazuhide , Suzuki Fujio 
Published: January 2023

Abstract
Hepatocellular carcinoma (HCC) is the fifth common cancer in the world. In previous study, we reported that M2b monocytes which predominately generated in advanced-stage HCC patients were important target for HCC immunotherapy. Recently, an anti-PD-1 antibody has been approved for the treatment against HCC. In this study, we tried to analyzed for immunological characters of monocytes isolated from patients with several stages of HCC.
From 2017 August to 2017 December, nine patients diagnosed as advanced-stage HCC patients and five patients diagnosed as early-stage HCC patients were enrolled in this study. Monocytes from peripheral blood of these patients were stained for anti-PD-1 antibody, anti-PD-L1 antibody and anti-PD-L2 antibody. These cells were analyzed for flow cytometry. Furthermore, five healthy donors were used for the control groups.
In the results, a majority of monocytes from advanced-stage HCC patients were expressed PD-L1 (81.5 ± 14.7%) and PD-L2 (71.6 ± 18.5%), but not PD-1(20.2 ± 18.6%). And, monocytes from early-stage HCC patients were expressed PD-L1 (9.9 ± 4.1%), PD-L2 (38.74 ± 9.9%) and PD-1 (23.5 ± 9.8%). As the contorls, monocytes from healthy donors were not expressed PD-L1 (21.7 ± 5.5%), PD-L2 (31.7 ± 23.8%) and PD-1 (11.6 ± 6.4%). The PD-L1 expression in monocytes was increased as HCC progressed.
These results indicate the PD-L1 expression may be used as a new marker for the classification of the cancer progression. And, PD-1−PD-L1+PD-L2+ monocytes which were predominantly generated in advanced-stage HCC patients, may target for the treatment of anti-PD-L1 antibody.
NOD-scid IL2rg
null (NSG) mice deficient in murine MHC Class I and Class II expression support engraftment of functional human T cells in the absence of acute xenogeneic GVHD following injection of PBMC
Authors: Brehm Michael A. , Wiles Michael , Kenney Laurie Lea , Burzenski Lisa , Keck James , Greiner Dale L. , Shultz Leonard D. 
Published: January 2023

Abstract
Humanized mice are emerging as an exciting platform to study human immuno-oncology and provide preclinical tools to test cutting edge immunotherapies. An effective and simplistic approach to engraft immunodeficient mice with functional human T cells is the injection of human peripheral blood mononuclear cells (PBMC). This PBMC engraftment model (termed Hu-PBL-SCID) has supported studies of human immune system-tumor interactions following injection of human PBMC into immunodeficient mice implanted with patient-derived (PDX) tumors. However, a major limitation of this model is the development of acute xenogeneic GVHD due to human T cell recognition of murine MHC class I and class II. To address this limitation, we created two strains of NSG mice that lack murine MHC antigens, one by crossing NSG-B2Mnullwith NSG-(IA IEnull) mice, the second by knocking out the MHC class II IAb gene in NSG-(KbDb)null mice using TALEN technology. We observed that human IgG clearance was rapid in NSG-B2Mnull(IA IEnull) mice that lack functional FcRn activity whereas clearance in NSG-(KbDb)null(IAnull) mice was comparable to that observed in NSG mice. Injection of human PBMC into both strains led to long term engraftment of human T cells without development of acute GVHD. The engrafted human T cells were functional as documented by their ability to reject human islet allografts. Moreover, NSG-(KbDb)null(IAnull) mice injected with a PDX tumor and PBMC from an allogeneic donor supported significant tumor growth in the absence of acute GVHD. These data document that NSG mice deficient in murine MHC class I and class II support studies of human immune system-tumor interactions in the absence of acute GVHD and provide a model to evaluate human antibody therapeutics.
The ceramide structure of sulfatide-analogues influences the functional activity of type II NKT cells
Authors: Pasquet Lise , Camara Kaddy , Bloom Anja C. , Richardson Stewart K. , Howell Amy R. , Terabe Masaki , Berzofsky Jay A. 
Published: January 2023

Abstract
The immune system plays a dual role in tumor elimination. Tumor antigens activate cytotoxic cells that kill tumor cells, but the autologous origin of tumors also activates regulatory cells supporting tumor escape. The CD1d-restricted sulfatide–reactive type II NKT cells have a regulatory role by increasing the development of lung metastasis after sulfatide injection in a murine model of colon carcinoma. The identification of compounds able to control the activation of type II NKT cells is essential.
Sulfatide (C24:1), a lipid enriched in many tumors, is composed of a galactose head with a sulfate group β-linked to a ceramide moiety. The structure of the ceramide influences the TCR/lipid-CD1d complex interaction and determines the reactivity to the lipid. We synthesized diverse sulfatide-analogues varying in the number of double bonds and hydroxyl groups in the ceramide. Using analogue-loaded-CD1d tetramers, we identified analogue-specific cells in the lung. These cells were also sulfatide-reactive but distinct from α-GalCer-specific type I NKT cells. In vitro, analogues exhibiting a phytosphingosine chain activated both type II and type I NKT cell hybridomas. All analogues induced CD1d-dependent IL-13 production by lung cells but in differing amounts. In vivo, analogues had variable effects on lung metastasis. Interestingly, the analogue C24:2, recognized only by the type II NKT hybridoma, induced significantly more IL-13 production by lung cells than sulfatide while significantly reducing the development of lung metastasis. The distinct effects of sulfatide and C24:2 on the specific activation of type II NKT cells and on tumor elimination could make C24:2 a valuable addition to the armamentarium of anti-tumor immunotherapies.
De novo DNA methylation programs restrain T cell rejuvenation during immune checkpoint blockade therapy
Authors: Youngblood Ben A. , Ghoniem Hazem , Moustaki Ardiana , Abdelsamed Hossam Aly , Fan Yiping , Thomas Paul G. , Federico Sara , Stewart Elizabeth 
Published: January 2023

Abstract
Immune-checkpoint blockade (ICB)-mediated rejuvenation of CD8 T cell effector functions has emerged as one of the most promising frontiers for treating cancer and chronic infections. However, antigen-specific T cells that have experienced prolonged antigen exposure are often terminally differentiated, and have a limited capacity to mount an effector response during ICB treatment. Such exhaustion of effector potential is a major impediment of current T cell based immunotherapy efforts. Using in vivo mouse models of tumor and chronic viral infection, we assessed the role of de novo epigenetic programming in establishing ICB-refractory exhausted T cells. We observed that genetic deletion of Dnmt3a in T cells at the effector stage of an immune response to chronic lymphocytic choriomeningitis virus (LCMV) infection allowed antigen-specific T cells to remain highly functional despite expressing high levels of PD-1 and having prolonged exposure to antigen. Quite strikingly, PD-1 blockade treatment of chronically infected animals resulted in massive expansion of PD-1+ Dnmt3a-deficient antigen-specific T cells. Whole-genome methylation profiling of WT and Dnmt3a-deficient LCMV-specific CD8 T cells identified de novo DNA methylation programs that are coupled to development of ICB-nonresponsive virus and tumor-specific T cells. Building upon these findings, we have identified de novo epigenetic programs acquired in human tumor-associated PD-1hi CD8 T cells. Collectively, these data establish Dnmt3a-mediated de novo DNA methylation programming as a key regulator in establishing ICB-refractory exhausted CD8 T cells and highlights epigenetic reprogramming of T cells as a novel approach to enhance T cell-based cancer therapies.
Glycogen synthase kinase-3 (GSK-3) inactivation compensates for the lack of CD28 in the priming of CD8+ cytotoxic T-cells: Implications for anti-PD-1 immunotherapy
Authors: Rudd Christopher E. , Krueger Janna , Taylor Alison 
Published: January 2023

Abstract
The rescue of exhausted CD8+ cytolytic T-cells (CTLs) by anti-programmed cell death-1 (anti-PD-1) immune checkpoint blockade has been found to require CD28 expression. At the same time, we have shown that the inactivation of the serine/threonine kinase glycogen synthase kinase (GSK)-3α/β with small-interfering RNAs (siRNAs) and small molecule inhibitors (SMIs) specifically down-regulates PD-1 expression for enhanced CD8+ CTL function and clearance of tumors and viral infections (Taylor et al., 2016 Immunity, 44(2):274-86, Taylor et al., 2017 Cancer Research, canres.0491.2017). It has been unclear whether the GSK-3α/β pathway accounts for CD28 costimulation of CD8+ CTL function. In this presentation, we show for the first time that the inactivation of GSK-3α/β through siRNA or by SMIs during priming can substitute CD28 co-stimulation in the potentiation of cytotoxic CD8+ CTL function against the EL-4 lymphoma cells expressing OVA peptide. The effect was seen using several structurally distinct GSK-3 SMIs and was accompanied by an increase in Lamp-1 and GZMB expression. Conversely, CD28 crosslinking obviated the need for GSK-3α/β inhibition in its enhancement of CTL function. Our findings support a model where GSK-3 is the central co-signal for CD28 priming of CD8+ CTLs in anti-PD-1 immunotherapy. Anti-PD-1 and GSK-3 SMIs cooperate synergistically to limit the growth of solid melanoma tumors in mice. We will present data on the altered nature of tumor-infiltrating lymphocytes (TILs) in response to anti-PD-1 versus GSK-3 blockade and combination therapy. Our findings outline a next-generation approach using small molecule inhibitors in combination with check-point blockade therapy for the elimination of tumors.
IL-33 inhibits tumor growth through the activation of anti-tumor CD8+ T cell response in hepatocellular carcinoma
Authors: Jin Ziqi , Liu Haiyan 
Published: January 2023

Abstract
IL-33 secreted by epithelial cells and activated innate immune cells is a member of IL-1 family with dual-immunological effects. It can induce both type 1 and type 2 immune responses and its function as an alarmin has been well demonstrated. However, the role of IL-33 in the development of hepatocellular carcinoma (HCC) is not clear. Here, we examined the function of IL-33 in various murine HCC models by hydrodynamically injecting either IL-33-expressing tumor cells or IL-33-expressing plasmids into the liver of tumor-bearing mice. Tumor growth was greatly inhibited by IL-33 expression. Similarly, in DEN-induced HCC model, we also demonstrated that IL-33 expression inhibited tumor development. This effect was shown to be dependent on extracellular IL-33 and ST2 expression, because it was diminished in ST2 KO mice. Further study showed there were increased percentages and numbers of CD8+ T cells in both spleen and liver in IL-33-expressing tumor-bearing mice. The percentages and numbers of activated and effector CD4+ and CD8+ T cells were also increased in the liver with IL-33 expression. Moreover, IFN-γ production of the CD8+ T cells was upregulated in both spleen and liver by IL-33. The cytotoxicity of CTLs against tumor cells was also enhanced. Depletion of CD8+ and CD4+T cells diminished the antitumor activity of IL-33, suggesting the anti-tumor function of IL-33 was T cell-dependent. In vitro IL-33 treatment could preferentially expand CD8+ T cells and promote their activation and IFN-γ production. Taken together, we demonstrated in murine HCC models that IL-33 could inhibit tumor development through its interaction with ST2 to promote anti-tumor CD8+ T cell response.
Discovery screen platform for co-modulators of T cell effector function against tumors
Authors: Kim Edward , Sekar Keerthana , Sakamoto Doreen , Yen David , Lee Fei , Bailis Julie , Ouyang Wenjun , Zheng Yan 
Published: January 2023

Abstract
A major goal for cancer immunotherapy is to target pathways that drive optimal anti-tumor efficacy. T cell checkpoint therapy aims to enhance T cell responses during priming and/or within tumor tissue, but the key drivers of efficacy in broad patient populations has remained elusive. Here we first sought to determine whether responsiveness to anti-PD1 therapy in established syngeneic murine tumors depends on recruitment of T cells from lymphoid organs during treatment, or if PD1 blockade on T cells already within tumors is sufficient to drive efficacy. We compared anti-PD1 efficacy on established MC38 tumors when FTY720 (S1PR antagonist that blocks lymphocyte egress from lymphoid organs) or vehicle control was administered from the start of treatment. We found that anti-PD1 efficacy did not require additional T cells migrating from lymphoid organs, suggesting that PD1 blockade can act directly on T cells that were already present within tumor tissue. We next examined the functionality of tumor-infiltrating T cell subsets during anti-PD1 treatment and found that cytokine production is enhanced in a population of PD1intermediate T cells, but not on PD1high T cells. Together, this suggests that PD1 checkpoint therapy is driven by amelioration of a fraction of PD1+ tumor-infiltrated T cells, and that additional pathways within established tumors may need to be co-targeted to drive optimal efficacy. We then developed an in vitro discovery screening platform to identify pathways that co-modulate human T cell effector function against PDL1+ cells under conditions that drive high levels of PD1 on T cells. We are currently screening genes of interest that are expressed in patient tumor tissues and highly associated with a T cell-inflamed signature.
TIGIT blockade prevents CTL and NK cell exhaustion and leads to tumor rejection in mice
Authors: Tian Zhigang 
Published: January 2023

Abstract
T-cell Ig and ITIM domain (TIGIT), is a co-inhibitory receptor expressed on both T cells and NK cells. Studies by our group as well as other groups have shown that TIGIT contribute to immune tolerance by inhibiting both T cell and NK cell-mediated immune responses through binding its ligand, CD155, on APCs or target cells. In this study, we showed that TIGIT was up-regulated on CTL and NK cells during tumor progression in several tumor mouse models. TIGIT deficiency delayed inoculated B16/F10 tumor growth in vivo. Blocking TIGIT signaling in vivo using monoclonal antibodies, in combination with Flt3L overexpression in vivo by gene delivery, reversed the exhaustion of anti-tumor effector cells, CTL and NK cells, in B16/F10 tumor-bearing mice and significantly suppressed pre-established B16/F10 tumor growth and metastasis, thereafter improving overall survival. We also showed that TIGIT blockade inhibited colon cancer, breast cancer and MCA-induced fibrosarcoma growth in vivo. These findings demonstrated that TIGIT contributes to tumor-induced immune exhaustion and might be a promising target for cancer immunotherapy.
Immune Checkpoints Acting as Gate-keepers of T Lymphocyte Self-renewal
Authors: Carrera Maria 
Published: January 2023

Abstract
Immune checkpoint inhibition represents a promising therapeutic modality for several previously incurable cancer syndromes. Why successful T cell responses are achieved in some patients but not others is incompletely understood. Because blockade of more than one inhibitory signal is now being investigated as approach to increase the chances of therapeutic success, we investigated how combined loss of PD-1 and LAG-3 signaling in CD8+ T cells would influence a well-characterized immune response. After challenge with Vaccinia virus, T cells from mice lacking both inhibitory pathways underwent a more vigorous primary effector cell response but were severely defective during a secondary challenge. During modeled T cell activation, immune checkpoint inhibition resulted in greater production of irreversibly differentiated effector cell progeny, but this came at the expense of accelerated depletion of self-renewing progenitor cells. Pharmacological interventions that impede anabolism-associated signaling were able to correct progenitor cells loss in the face of checkpoint inhibition. Unleashing greater and greater T cell activation may result in regenerative failure of critical clones. By fostering the expansion of self-renewing progenitors, reversible agents that dampen T cell signal strength might paradoxically improve durability of immune checkpoint inhibition.
CD57+ T cells phenotype and function in non-small cell lung cancer
Authors: Li Lequn , Huang Bing 
Published: January 2023

Abstract
The composition of TILs is heterogeneous and the contribution of various subsets of TILs to anti-tumor effects can be incredibly different. Therefore, understanding the immune function of existing subsets of TILs in particular cancer types is of great significance. Previous studies, conducted by others and ourselves, have shown that CD103+ TILs have good prognostic value and are functionally primed to respond to PD-1 blockade. In this study, we investigated the phenotypic and functional features of CD57+ TILs in non-small cell lung cancer (NSCLC). CD57+T cells have been considered as end-stage, senescent T cells during chronic viral infections. However, the immune functions of CD57+ T cells in NSCLC remain unclear. Using a multicolor staining approach, we analyzed TILs isolated from freshly resected tumor tissues from NSCLC patients and determined that CD57+ T cells were largely CD8+ cells and were a different subset from CD103+T cells. CD28 expression was significantly downregulated in CD57+ T cells compared to CD57−T cells. Functional analysis revealed that CD57+ T cells produced a significant amount of IFN-γ in response to PMA/IONMYCIN stimulation compared to CD57− T cells, suggesting that these cells are functional effector/effector memory cells within the tumor microenvironment. Interestingly, immune response of CD57+ T cells in vitro was significantly impaired as determined by the expression of IFN-γ and Ki-67 in response to anti-CD3/CD28 stimulation. In contrast to CD103+ T cells, additional PD-1 inhibitor did not improve the ability of CD57+ T cells to produce IFN-γ. These results suggest that CD57+ T cells combined with CD103+ T cells might serve as predictive biomarkers for PD-1 based immunotherapy.
PD-1 blockade activates conventional CD4 T cells and the innate immune response during glioblastoma eradication
Authors: Klein Sarah , Ziello Jennifer , Speranza Maria , Gokhale Prafulla , Kirschmeier Paul , Crosby Katherine , Freeman Gordon , Reardon David 
Published: January 2023

Abstract
Blockade of immune cell co-inhibitory receptor PD-1 using monoclonal antibodies enables anti-tumor immune responses in various solid tumors and lymphoid malignancies. Our laboratory previously demonstrated PD-1 blockade elicits an anti-tumor immune response resulting in tumor rejection and long-term survival in approximately 50% of mice with an orthotopic GL261 glioblastoma, despite lacking a corresponding accumulation of CD8+ cytotoxic T cell in the tumor or draining lymph nodes. In this investigation, we evaluated the role of conventional CD4+ T cells and the innate immune response in PD-1 mediated anti-glioma immunity using multiplex technologies for immunohistochemistry and flow cytometry. In response to PD-1 monotherapy, intratumoral CD4 T cells expressed significantly elevated levels of proteins required for T cell proliferation, activation, and effector function. CD4 T cell activation was accompanied by the classical activation and M1 polarization of resident microglia and tumor-infiltrating macrophages, including down-regulation of PD-L1 and up-regulation of MHC class II surface expression. We also demonstrated that depletion of either CD4 or CD8 T cells was sufficient to completely ablate anti-PD-1-mediated tumor eradication and long-term survival. Our data suggests CD4 T cells and myeloid cells may play a prominent role in the eradication of glioblastoma by PD-1 blockade.