bluuebunny/crossref_metadata_2025_split huggingface dataset:
BBB-permeable peptide conjugated cytoplasmic domain of CTLA-4 inhibits Th1 and Th17 responses and pathogenesis of multiple sclerosis
Authors: Lim Sangho , Kim Won-Ju , Kim Yeon-Ho , Lee Sohee , Koo Ja-Hyun , Lee Jung-Ah , Kim Hye-Mi , Park Hong-Jai , Kim Do-Hyun , Lee Hong-Gyun , Yoon Heeseok , Kim Ji Yun , Shin Jae Hun , Kim Lark Kyun , Doh Junsang , Kim Hongtae , Bothwell Alfred L , Lee Sang-Kyou , Suh Minah , Choi Je-Min 
Published: January 2023

Abstract
Multiple sclerosis (MS) is one of the most severe autoimmune disease, which cause severe inflammation in central nervous system (CNS). In the MS, CNS-infiltrating effector T cells are regarded that play critical roles. However, current drugs for MS could not targeting infiltrated T cells due to the limitations of drug delivery, which cannot penetrate blood-brain barrier (BBB) and deliver into the CNS. Here, we identified a novel BBB-permeable peptide, dNP2, which effectively delivered proteins into the CNS in vivo. It transduced cargo proteins into the cells with higher efficiency than other cargo delivery peptides. Moreover, it localized in resident neuron, astrocytes and microglia of the mouse brain through blood vessels by penetrating the BBB. Also, we found that dNP2 can deliver its cargo protein into the infiltrated T cells in the CNS of MOG35-55 immunized MS model mice. When we treated the dNP2-conjugated cytoplasmic domain of cytotoxic T lymphocyte antigen 4 (dNP2-ctCTLA-4), it efficiently downregulated cytokine production in activated T cells, and showed inhibitory impact on experimental autoimmune encephalomyelitis (EAE) in both preventive and therapeutic schemes. This was accompanied with reductions of demyelination and infiltration of T helper 1 (Th1) and T helper 17 (Th17) cells in the CNS. This study suggests that dNP2 is a novel BBB-permeable peptide and that dNP2-ctCTLA-4 could be an effective agent for treating CNS inflammatory diseases such as MS.
IL-33 is required for kidney ischemia-reperfusion-induced injury in mice
Authors: Herbelin Andre , Ferhat Maroua , Giraud Sébastien , Robin Aurélie , Goujon Jean-Michel , Hauet Thierry , Gombert Jean-Marc , Thierry Antoine 
Published: January 2023

Abstract

Introduction
Ischemia-reperfusion (IR) injury during kidney transplantation induces necrosis of renal cells and the release of “alarmins” such as IL-33 that can activate the innate immune system, thereby triggering an inflammatory response and tissue damage leading to renal failure, dysfunction or rejection. The aim of this study was to investigate the contribution of IL-33 in kidney IR injury.

Methods
10–12-week-old wild-type (WT) and IL-33-deficient (IL-33Gt/Gt) male C57/Bl6 mice were subjected to 32 minutes of unilateral kidney ischemia or a Sham operation. After 24 hours, kidneys were harvested and leucocyte infiltration (macrophages, neutrophils, NK and NKT cells) was analyzed by flow cytometry. Renal injury was assessed by measurement of plasma creatinine and histological grading.

Results
Plasma creatinine level and tissue damage significantly increased after renal IR in WT mice, as compared with Sham-operated animals, a difference that disappeared when WT mice were replaced by IL-33Gt/Gt mice. Even though intra-renal neutrophil (CD11b(+)GR-1(+)) were significantly increased 24 hour-post IR in both WT and IL-33Gt/Gt mice, this phenomenon was found to be significantly attenuated in IL-33Gt/Gt mice. Moreover, monocyte/macrophages (CD11b(+)F4/80(+)) and NKT cells, also known for their deleterious effect during renal IR injury, seemed to be less recruited 24 hour-post IR in IL-33Gt/Gt mice compared with WT mice.

Conclusion
IL-33Gt/Gt mice are less sensitive to kidney damage 24 hour-post IR, consistent with a deleterious effect of IL-33 during renal IR injury. This study underlines a new possible role of IL-33 as an innate-immune mediator during kidney IR injury.
NK Cells Play a Critical Role in Mediating Inflammation and Graft Failure During Antibody-Mediated Rejection of Kidney Allografts
Authors: Fairchild Robert L , Kish Danielle , Tanaka Toshiaki , Kohei Naoki , Dvorina Nina , Valujskikh Anna , Baldwin William M. , Fairchild Robert 
Published: January 2023

Abstract
The incidence of antibody-mediated kidney graft rejection has increased, but the key cellular and molecular participants underlying this graft injury remain unclear. We previously reported that kidney allograft rejection in CCR5−/− mice is dependent on donor-specific antibody (DSA). The current study determined if cells expressing cytotoxic function contributed to antibody-mediated kidney allograft rejection in CCR5−/− recipients. Wild type C57BL/6, B6.CCR5−/− and CD8−/−/CCR5−/− mice were transplanted with complete MHC mismatched A/J kidney grafts and intra-graft inflammatory components were followed to rejection. B6.CCR5−/− and CD8−/−/CCR5−/− recipients rejected the allografts by day 35 whereas 65% of allografts in wild type recipients survived past day 80 post-transplant. Allograft rejection in wild type recipients was associated with high titers of DSA, equivalent to those induced in CCR5-deficient recipients. Rejected allografts in C57BL/6, B6.CCR5−/− and CD8−/−/CCR5−/− recipients’ expressed high levels of VCAM-1 and MMP7 mRNA that was associated with high serum titers of DSA. At rejection in wild type and CD8−/−/CCR5−/− recipients, kidney allografts also expressed genes associated with NK cell (Sh2d1B1 and MYBL1) but not with T cell (CXCR6) activity during inflammation. High levels of perforin and granzyme B mRNA expression in kidney allografts peaked on day 6 post-transplant in all recipients. NK cell depletion in CD8−/−/CCR5−/− recipients reduced this expression to background levels and promoted long-term survival of 40% of kidney allografts. These results support a role for NK cells in increasing inflammation during antibody-mediated kidney allograft injury and rejection.
Suppression of Tumorigenicity 2 (ST2) signaling is required for regulatory T cell control of Graft-vs. Host Disease
Authors: Zhang Xiaoli , Matta Benjamin M , Reichenbach Dawn K , Blazar Bruce R , Turnquist Heth R 
Published: January 2023

Abstract
How IL-33-mediated signals impact on ST2-expressing regulatory T cell (Treg) expansion and regulatory functions is poorly understood. To establish the importance of direct IL-33 stimulation of Treg for control of alloimmune responses, we utilized a rodent model of graft-vs. host disease (GVHD) where C57BL/6 mice were irradiated and given a BALB/c bone marrow transplant (BMT) along with BALB/c CD4+ CD25+ Treg from st2+/+ or st2−/− mice at a 1:2 ratio with WT BALB/c CD3+ effector T cells. As expected, WT st2+/+ CD4+ CD25+ cells promoted full protection against GVHD with 90% of recipients receiving st2+/+ CD4+ CD25+ free of GVHD at Day 100 post-BMT. However, only 10% of mice that received st2−/− Treg achieved long-term survival (p=0.0017, st2+/+ v. st2−/−). Associated mechanistic studies revealed that Treg frequency was significantly reduced in recipients receiving st2−/− Treg compared to those receiving st2+/+ Treg. To define the signaling patterns IL-33 generates in Treg, fluorescence activated cell sorting was used to obtain ST2− or ST2+ Treg cells from the splenocytes of Foxp3 reporter mice and IL33 activated signaling downstream of ST2 was quantitated by phosphoflow cytometry. This analysis revealed that IL-33 signaling via ST2 on Treg activates both NF-κB and p38 MAPK. Yet, only IL-33-stimulated p38 MAPK, but not NF-kB, signaling was required for ST2+ Treg expansion. Our studies make two important observations regarding how IL-33 stimulation of Treg supports immune regulation. First, we demonstrate that IL-33 activates p38 MAPK to drive selective ST2+ Treg expansion. Second, we show that IL-33-mediated signals are critical to the capacity of Treg to control the alloimmune responses leading to GVHD.
Development of an Enhanced VETO cells for the generation of Alloantigen-specific Tolerance
Authors: Rajawat Yogendra Singh , McFarland Hugh , Morgan Richard , Zhao Yangbing , Rosenberg Amy S 
Published: January 2023

Abstract
Development of specific immunotherapies to prevent graft rejection and graft-versus-host disease offers the promise of selectively deleting only allospecific immune responses. Veto activity is the ability of a cell to specifically suppress/delete only T cells directed against antigens of the veto cells themselves, but not against third-party antigens. In the present study, we reasoned that providing TCR-like signaling to allogeneic major histocompatibility complex (MHC) molecules on the surface of CD8+CTLs might result in enhanced veto function. To generate the donor antigen specific veto cells, we have utilized the Chimeric Antigen Receptor (CAR) T cell approach, in which CD8+CTLs express an MHC alloantigen fused to signaling molecule apparatuses. We fused mouse MHC class I alloantigen (H-2Dd) to the hinge and trans-membrane domains of the mouse CD8 chain plus intracellular signaling sequences derived from mouse 4-1BB, and CD3-ζ chain creating various CAR constructs. These CARs were cloned into retroviral vectors. Ex-vivo transduced CD8+T cells (FVB-Dd) were adoptively transferred in the recipient parental mice strain (FVB-H-2q). 4–6 weeks post adoptive transfer, we detected low levels of Dd+ CD8+T cells in lymph node and spleen, ranging from 2–3% of total CD8+T cells. Dd expressing skin grafts from 3604 transgenic mice (H-2Dd transgenic mice on FVB-H-2q background) were transplanted on recipient mice and preliminary data suggests that skin graft survival was extended for 4 months (usual rejection in naïve mice is by 15–18 days).Veto activity of these Dd+CD8+T cells will be examined by in vitro Dd specific CTL assays, proliferation assays.
Protein Kinase C-θ and vimentin modulate multiple facets of Regulatory T-cell function.
Authors: McDonald-Hyman Cameron , Thangavelu Govindarajan , Saha Asim , Muller James , Zhang Guoan , Kumari Sudha , Koehn Brent H. , Mitchell Jason S. , Fife Brian T , Serody Jonathan S , Osborn Mark J , Hippen Keli L , Kelekar Ameeta , Munn David H , Altman Amnon , Neubert Thomas , Dustin Michael L , Blazar Bruce R 
Published: January 2023

Abstract
Regulatory T-cells (Tregs) prevent autoimmune and alloimmune reactions. Augmenting Treg function may enhance Treg therapies for these diseases. Treg-specific inhibition of protein kinase C-theta (PKC-θ) enhances Treg function. However, it is unclear whether PKC-θ inhibition can boost Treg function in systemic inflammatory conditions.
In a mouse model of acute GVHD, we found that Tregs treated with the PKC-θ inhibitor AEB071 reduced GVHD mortality and severity significantly better than DMSO treated Tregs. Compared to DMSO, AEB071 treated Tregs significantly reduced conventional T-cells (Tcon) proliferation on D4 after transplant. Multi-photon microscopy showed that AEB071 treated Tregs significantly increased Tcon velocity and displacement compared to DMSO.
Mechanistically, AEB071 augments expression of Neuropilin-1 (Nrp1) and Lymphocyte activation gene 3 (Lag3). Antibody blockade of Nrp1 and Lag3 in transwell suppression assays reduced the effect of AEB071 on Treg function. PKC-θ inhibition also reduces phosphorylation of mTORC2 targets FoxO3a and Akt phospho-site S473, but not mTORC1 targets S6, 4E-BP1 or Akt phospho-site T308. Compared to DMSO, AEB071 treatment significantly increased fatty acid uptake and oxygen consumption rate (OCR).
Phosphoproteomic analysis identified a significant alteration in the interaction between PKC-θ and the intermediate filament vimentin after AEB071 treatment, which was confirmed by confocal. Vimentin siRNA treatment also significantly reduces PKC-θ/vimentin interaction, increases Treg function, Nrp1 expression and OCR.
In summary, PKC-θ and vimentin modulate multiple aspects of Treg function, and altering these molecules may enhance the efficacy of Treg therapeutics.
Targeting PI3K signaling to ameliorate chronic graft versus host disease.
Authors: Paz Katelyn G , Flynn Ryan , Du Jing , Dong Shuai , Okkenhaug Klaus , Stark Anne-Katrien , Vanhaesebroeck Bart , Johnson Amy , Tannheimer Stacey , Turka Lawrence A , Byrd John , Queva Christophe , Blazar Bruce R 
Published: January 2023

Abstract
The phosphoinositide 3-kinase (PI3K) pathway is a key signaling pathway necessary for T cell activation, differentiation and metabolism. T effector cells rely on increased PI3K signaling to fuel glycolysis for their metabolic needs, while T regulatory cells downregulate PI3K and favor lipid oxidation. The metabolic processes of lymphocytes modulating chronic graft versus host disease (cGVHD) have remained largely unexamined and represent a novel therapeutic strategy for this disease. Here, we investigated the role of PI3K signaling in a murine model of cGVHD that is etiologically linked to up regulated germinal centers (GCs) and characterized by multisystem organ disease; including fibrosis of the lung, which results in pulmonary dysfunction. We hypothesized that inhibition of PI3K signaling would alter the activation and/or function of GC-facilitating T follicular helper (TFH) cells resulting in lessened disease. The findings in this study are that mice treated with a PI3Kd inhibitor had decreased pulmonary dysfunction similar to that of the control, non-cGVHD mice. The frequencies of splenic TFH cells as well as GC B cells were decreased by a PI3K delta inhibitor compared to non-treated cGVHD controls. In a similar manner, mice that received PI3K kinase delta dead Tregs also had decreased TFH frequency as well as reduced pulmonary dysfunction. Our results indicate the differential requirement for signaling through PI3K delta and suggest that targeting this pathway may be a potential new therapy for treatment of cGVHD. Additional studies are required to validate the potential therapeutic use.
Comprehensive pipeline for bioinformatics prediction of immunogenic minor histocompatibility antigens
Authors: Efimov Grigory , Bykova Nadezda , Vdovin Alexander , Filkin Serguei , Postovskaya Anna , Malko Dmitry 
Published: January 2023

Abstract
Hematopoietic stem cell transplantation (HSCT) is widely used for treatment of haematological malignancies. Both raft versus tumor (GVT) effect and potentially lethal graft versus host disease (GVHD) may develop after transplantation. Current approach to limit GVHD, is based on donor selection and immunosuppression. But as alloreactivity is not restricted to the HLA locus, up to 40% of fully HLA-matched transplantations are complicated by severe forms of GVHD. Non-synonymous single nucleotide polymorphisms (SNPs) in the genome coding regions can give rise to polymorphic peptides – minor histocompatibility antigens (MiHA) – recognized by the immune system as foreign.
Limiting alloreactivity to SNPs in hematopoietic genes could allow to discriminate between GVHD and GVT. Thus information of which polymorphisms are immunogenic is of great importance.
So far only a limited number of MiHA have been discovered. Methods for verification of SNP immunogenicity are slow and laborious so there is a need for algorithms predicting potentially immunogenic polymorphisms.
We have developed TIE predict (TransImmuno Epitope predictor) which is a comprehensive pipeline for in silico prediction of MiHAs based on exome data of donors and patients. This pipeline utilizes several algorithms for peptide to MHC binding prediction, which increases the overall accuracy of prediction. TIE predict is linked to TransImmuno Epitope database holding the data on common genetic poymophisms genomic sequences and annotations. We have tested this approach on publically available genetic data and currently starting to use it on patients and donors undergoing HSCT in combination with immunogenicity assays.
Group 3 NCR+ Innate Lymphoid Cells accumulate in human lung cancer
Authors: Ferlazzo Guido , Mingari Mariacristina , Carrega Paolo 
Published: January 2023

Abstract
Tertiary lymphoid structures (TLS) are a common finding in non-small-cell lung (NSCL) cancers and are predictors of favorable clinical outcome. Here, we show that NCR+ Innate-Lymphoid-Cells (ILC)3s are present in the lymphoid infiltrate of human NSCL carcinomas and are mainly localized at the edge of tumor-associated TLS. This intra-tumoral lymphocyte subset is endowed with lymphoid tissue-inducing properties and, upon activation, produces IL-22, TNF-α, IL-8, IL-2 and activates endothelial cells. Tumor-NCR+ ILC3 may interact with both lung tumor cells and tumor-associated fibroblasts, resulting in the release of cytokines primarily upon engagement of the NKp44 activating receptor. In patients, NCR+ ILC3s are present in significantly higher amounts in stage I/II NSCL cancers than in more advanced tumor stages. Our results indicate that NCR+ ILC3s are associated with human NSCL cancer and may contribute to the formation of protective tumor-associated TLS.
IP3 3-Kinase B Suppresses B Cell Lymphoma by Antagonizing PI3K/mTOR in B cells
Authors: Sauer Karsten , Conche Claire , Deng Yisong , Jin Hyun-Yong , Bethel Kelly , Rigaud Stephanie M , Westernberg Luise , Siegemund Sabine , Xiao Changchun 
Published: January 2023

Abstract
Accounting for ~4% of all cancers in the US, Non-Hodgkin Lymphoma (NHL) is the most prevalent blood cancer. Diffuse large B-cell Lymphoma (DLBCL) is the most common and aggressive type of NHL. Here, we identify inositol-trisphosphate (IP3) 3-kinase B (Itpkb) as a novel tumor suppressor whose deficiency triggers DLBCL in mice. We found that aging Itpkb−/− mice die prematurely with anemia and splenomegaly. 15% of these mice showed multiorgan-infiltration with neoplastic germinal-center phenotype B cells reminiscent of DLBCL. Itpkb−/−B cell lymphomas are mono-or oligoclonal, transplantable, and constitutively hyperactivate the oncogenic phosphoinositide 3-kinase (PI3K) signaling pathway. Growth of Itpkb−/−B cell lymphoma xenografts in immunodeficient mice is sensitive to treatment with PI3K or mTOR inhibitors. In humans, subsets of DLBCL patients carry Itpkb missense mutations, deletions or copy number reductions. Additionally, Itpkb expression is altered in several other hematopoietic and non-hematopoietic human cancers. We propose that Itpkb suppresses tumors by producing inositol-tetrakisphosphate (IP4), a soluble analog of the PI3K product phosphatidylinositol-trisphosphate (PIP3). We and others have previously shown that IP4 antagonism with PIP3 for Akt effector kinase binding prevents excessive Akt/mTORC1 signaling in hematopoietic cells. Our new results suggest that this mechanism prevents oncogenic PI3K signaling in the B cell lineage and possibly in other cell types. Thus, Itpkb might be a novel biomarker for tumor aggressiveness or prognosis, and Itpkb activating drugs or IP4 might have therapeutic potential.
IFNγ has variable effects on expression of MHC Class I and ICAM-1 in pediatric cancers which alter sensitivity to NK cell mediated lysis
Authors: Aquino-Lopez Arianexys , Senyukov Vladimir , Romain Gabrielle , Varadarajan Navin , Lee Dean A. 
Published: January 2023

Abstract
Interferon-γ (IFNγ), a cytokine secreted by effector lymphocytes upon activation, is known to cause upregulation of MHC class I in tumor cells, which has been associated with natural killer (NK) cell inhibition. NK cells are very heterogeneous with respect to receptor expression and their function depends on an interplay between these inhibitory and activating signals. In order to optimize NK cell therapy, it is essential to identify the NK cells expressing the optimal combination of receptors for a given target. In this study we evaluated tumor cell lines from the Pediatric Preclinical Testing Program to determine the impact of IFNγ on their expression of NK cell activating and inhibitory ligands, death receptors, and adhesion molecules using mass cytometry, and their correlation with tumor lysis. We found that IFNγ treatment can significantly increase ICAM-1 and/or HLA-ABC levels together or independently. When HLA-ABC increases without concomitant ICAM-1 upregulation, this is associated with decreased lysis. Conversely, when ICAM-1 upregulation exceeds HLA-ABC upregulation this is associated with increased conjugate formation and better NK cell cytotoxic activity. This increase in cytotoxicity and conjugate formation is lost after blocking ICAM-1. Time-lapse imaging of the NK cell interaction with tumor cells shows that NK cells require less time to form stable contacts with IFNγ treated tumor cells when compared to untreated tumor cells. This suggests that IFNγ induced upregulation of ICAM-1 enhances NK cell cytotoxic activity by reducing the time required for NK cells to form stable contacts with their target.
NKG2D receptor activation makes CD8+ T cells resistant to the suppressive effects of TGF-β
Authors: Prajapati Kushal , Perez Cynthia , Burke Brianna , Plaza-Rojas Lourdes , Guevara-Patino Jose Alejandro 
Published: January 2023

Abstract
Natural killer group 2, member D (NKG2D) is a stimulatory receptor majorly present on the surface of natural killer and CD8+ T cells. We and others have shown that NKG2D receptor enhances the function, persistence and proliferation of CD8+ T cells. On the other hand, a major hurdle in the field of cancer vaccine and adoptive T cell transfer therapy (ACT) is diminished effectiveness and persistence of CD8+ T cells due to immuno-suppressive factors. TGF-β is a highly prevalent immuno-suppressive factor that negatively regulates the function and persistence of CD8+ T cells. We hypothesized that NKG2D activation opposes the suppressive effects of TGF-β on CD8+ T cells, and enhances their anti-tumor activity in tumor microenvironment. Here, we show that activating the NKG2D receptor makes human CD8+ T cells resistant to TGF-β’s suppressive effects on the activation, proliferation and granzyme B expression. Further, we found that NKG2D knock-out mouse CD8+ T cells are more susceptible to these TGF-β mediated suppressive effects in vitro. Interestingly, under influence of TGF-β, NKG2D KO CD8+ T cells’ ability to secrete multiple cytokines in vivo was significantly less than their WT counterparts. Our next objective is to determine the molecular mechanisms by which NKG2D makes CD8+ T cells resistant to TGF-β. For the proximal mechanism, we will determine NKG2D’s immediate signaling pathway, namely PI3K or Grb2-SOS-Ras, mediating resistance to TGF-β. Moreover, our preliminary data shows that NKG2D activation increases the levels of regulator of G protein signaling 3 (RGS3), inhibitor of TGF-β signaling, in CD8+ T cells. We will investigate if up-regulation of RGS3 is a distal mechanism underlying NKG2D mediated resistance to TGF-β in CD8+ T cells.