bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolite-Based Microparticles Enhance the Metabolic Fitness of T cells
Authors: Wu Christopher , Inamdar Sahil , Acharya Abhinav P. , Curtis Marion 
Published: February 2023

Abstract
Adoptive Cell Transfer (ACT) therapies have recently fostered excitement for the treatment of hematogenous cancers. However, efficacy has largely been stunted in the treatment of solid tumors, especially for ovarian cancer, in part due to nutrient restriction within the tumor microenvironment. We recently demonstrated that intracellular delivery of biodegradable microparticles (MPs) made from polymers of alpha-ketoglutarate modulated dendritic cell energy metabolism and immunophenotype, altering regulatory and helper T cell frequencies. To determine if continuous metabolite release from MPs would rescue T cell dysfunction in nutrient-poor, immunosuppressive conditions, we generated MPs of polymerized glycolytic and tricarboxylic acid cycle central-carbon metabolites. Flow cytometric analysis and confocal microscopy indicated that FITC-conjugated MPs were efficiently loaded into the cytoplasm of activated primary murine T cells via membrane adsorption or electroporation. The MPs were retained intracellularly and sustained proliferation of mouse T cells for over five days in vitro. Glycolysis and mitochondrial respiration stress tests using an Agilent Seahorse XFe96 Analyzer demonstrated that primary mouse CD8 T cells utilized the MPs as a metabolic substrate. In particular, MPs increased metabolic function, as indicated by increased spare respiratory capacity, glycolysis, and glycolytic capacity. The results of this study provide preliminary evidence in support of utilizing sustained-releasing particles of metabolic substrates for the metabolic reprogramming of immune cells used in ACT therapies. This strategy has potential to improve or expand the efficacy of existing ACT therapies for solid tumors.
Analysis of soluble immune checkpoint proteins using quantitative multiplex microbead-based immunoassays
Authors: Lie Wen-Rong , McDonald John , Russell Donna , Banerjee Munmun , Zimmermann Daniel , Kornmeier Christine , Pepin Danielle 
Published: February 2023

Abstract
Immune-checkpoint receptors play critical roles in immune regulations and maintenance of immune homeostasis. Recently, a series of soluble immune checkpoint proteins have been found in circulation, and are putative biomarkers in infection, cancer, and autoimmune diseases. We have developed two multiplex immuno-oncology (I/O) panels. Human I/O Checkpoint Protein Panel 1 (17-plex) consists of 17 targets: BTLA, CD27, CD28, CD40, CD80, CD86, CTLA-4, GITR, GITRL, HVEM, ICOS, LAG-3, PD-1, PD-L1, PD-L2, TIM-3, TLR-2. Human I/O Checkpoint Protein Panel 2 (31-plex) consists of 31 targets: 4-1BBL, CD73, APRIL, Arginase-1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, BAFF, CD25, CD30, CD40L, 4-1BB, CD226, E-Cadherin, FGL1, Gal-1, Gal-3, Granulysin, Granzyme B, IDO1, MICA, MICB, Nectin-2, Nectin-4, OX40, Perforin, CD155, Siglec-7, and Siglec-9. Using these multiplex arrays, we have simultaneously quantitated the expression levels of the above 48 key immune regulators in cancer serum samples and in cell culture supernatants. Soluble checkpoint protein signatures generated from this multiplex approach revealed differential expression of sBTLA, sCD27, sCD40, sCD86, sHVEM, sPD-1, sTIM-3, sTLR-2, sCD40L, Gal-3, Gal-1, FGL1, BAFF in the colorectal or breast cancer serum samples, compared to healthy serum controls. Heatmaps demonstrate the detection of checkpoint proteins in the conditioned media collected from human tumor cell lines and stimulated PBMCs. In summary, our results demonstrate that these two multiplex immunoassays we developed are useful research tools for the simultaneous quantitation of circulating immune checkpoint proteins, as well as for its potential application in biomarker discovery and translational research.
Identification and function of a CD4+/CD8αβ+ T cell population that is predictive of GVHD development in a xenogeneic transplant model
Authors: Hess Nicholas J , Hematti Peiman , Gumperz Jenny E , Capitini Christian M. 
Published: February 2023

Abstract
Graft-vs-host disease (GVHD) is mediated by donor reactive T cells that have a hierarchical classification based on CD4 and CD8 expression. While CD4 and CD8 lineages are thought to have fixed expression, CD4+/CD8αβ+ double positive (DP) T cells have been reported in cases of human cancers and autoimmune diseases though the lack of a suitable model system has hindered their research. In this study, we transplanted primary human graft tissue into non-conditioned immunodeficient mice and observed the development of a human DP T cell population that was not present in the starting grafts. This DP T cell population developed irrespective of graft tissue (peripheral blood, bone marrow or umbilical cord blood), accessory cells (transplantation with isolated T cells) and immunodeficient mouse strain (NSG and NBSGW). Furthermore, an increase in the percentage of DP T cells in the blood of these mice is correlated and predictive of GVHD development. We also observed that DP T cells are functionally active with significantly elevated IFNγ and TNFα secretion compared to CD4 and CD8 single positive T cells. DP T cell also display elements of the cytotoxic machinery including NKG2D and perforin/granzyme expression. Interestingly, transplantation of isolated CD4+ cells did not result in the development of DP T cells while a robust population developed after transplantation of isolated CD8+ T cells. DP T cells were also identified in primary clinical samples taken from HSCT patients with their clinical relevance to GVHD currently under investigation. In conclusion, this ongoing study has identified a novel human DP T cell that arises from the CD8+ T cell population, is functional active and is predictive of GVHD in a xenogeneic transplant model.
Transplantation and CMV promote premature aging of CD8 T cells
Authors: Higdon Lauren E , Gustafson Claire E , Ji Xuhuai , Maecker Holden T , Maltzman Jonathan S 
Published: February 2023

Abstract
Cytomegalovirus (CMV) infection is a significant contributor to both immune aging and complications after solid organ transplantation. A major mediator of both effects is viral reactivation, in which latent CMV in the host undergoes new replication requiring immune control. Immunocompetent CMV seropositive individuals display hallmarks of CD8 T cell aging earlier than CMV seronegative individuals, including shortened telomeres, defects in proliferation and signaling, and altered cytokine production. In the context of transplantation, immunosuppression impairs immune control, leading to negative impacts including late-term graft loss. We hypothesized that T cell aging is accelerated after transplant in CMV+ individuals, regardless of detectable viral reactivation. To address this hypothesis, we measured telomere length and conducted phenotyping through flow cytometry and targeted single cell RNA sequencing. We found a statistical increase in CD57 expression which correlated with a trend towards decreased telomere length in the first year after transplantation. We found that CD57+ CD8 T cells expanded in the first year after transplantation specifically in CMV+ recipients. We also found that clonal expansion was associated with a terminally differentiated phenotype, defined as CD45RA+ CD27− CCR7−. Overall, our data demonstrate an enhanced aging phenotype in the first year post-transplant in CMV seropositive individuals, based on CD57+ expression and terminal differentiation. These findings suggest that CD8 T cell aging may be an important factor to understanding immunity in transplant recipients.
DNA-PKcs Inhibition Extends Allogeneic Skin Graft Survival
Authors: Burdine Marie , Waldrip Zachary , Burdine Lyle 
Published: February 2023

Abstract

Background:
Organ transplantation is life-saving and continued investigations into immunological mechanisms that drive organ rejection are needed to improve immunosuppression therapies and prevent graft failure. DNA-dependent protein kinase catalytic subunit, DNA-PKcs, is a critical component of both the cellular and humoral immune responses. In this study, we investigate the contribution of DNA-PKcs to allogeneic skin graft rejection to potentially highlight a novel strategy for inhibiting transplant rejection.

Methods:
Fully MHC mismatched murine allogeneic skin graft studies were performed by transplanting skin from BalbC mice to C57bl6 mice and treating with either vehicle or the DNA-PKcs inhibitor NU7441. Graft rejection, cytokine production, immune cell infiltration, and donor-specific antibody (DSA) formation were analyzed.

Results:
DNA-PKcs inhibition significantly reduced necrosis and extended graft survival compared to controls (mean survival 14 days vs 9 days respectively). Inhibition reduced the production of the cytokines Interleukin (IL)2, IL4, IL6, IL10, TNFα, and IFNγ and the infiltration of CD3+ lymphocytes into grafts. Furthermore, DNA-PKcs inhibition reduced the number of CD19+ B cells and CD19+ CD138+ plasma cells coinciding with a significant reduction in DSAs. At a molecular level, we determined that the immunosuppressive effects of DNA-PKcs inhibition were mediated, in part, via inhibition of NFκB signaling through reduced expression of the p65 subunit.

Conclusion:
Our data confirm that DNA-PKcs contributes to allogeneic graft rejection and highlight a novel immunological function for DNA-PKcs in the regulation of NFκB and concomitant cytokine production.
Cell death stage dictates the immunogenicity of ferroptotic cancer cells
Authors: Efimova Iuliia , Catanzaro Elena , Demuynck Robin , Van der Meeren Louis , Coppieters Frauke , Bachert Claus , Skirtach Andre , Krysko Olga , Krysko Dmitri 
Published: February 2023

Abstract
Immunotherapies hold great promise for the future treatment of cancer. It is also becoming clear that type of cancer cell death determines the antitumor immune response and, therefore, contributes to the efficiency of anti-cancer therapy and long-term survival of patients. Since tumors often develop resistance to apoptosis and necroptosis, triggering these processes is not always the optimal strategy. That is why it is crucial to select the proper cell death type potentially leading to tumor eradication. Ferroptosis, an iron-dependent form of cell death leading to lipid peroxidation in cells, is currently actively studied but whether ferroptotic cancer cells are immunogenic is unknown. We demonstrate for the first time that ferroptosis is immunogenic in vitro and in vivo. Early, but not late, ferroptotic cells promote the phenotypic maturation of BMDCs and elicit a vaccination-like effect in immune-competent mice but not in Rag-2−/− mice, suggesting that the mechanism of immunogenicity is very tightly regulated by the adaptive immune system and is time dependent. Also, ATP and HMGB1, the best-characterized damage-associated molecular patterns involved in immunogenic cell death, have proven to be passively released along the timeline of ferroptosis and act as immunogenic signal associated with the immunogenicity of early ferroptotic cancer cells. The work on approaches to increase the immunogenicity of late ferroptotic cells is underway, which will open up novel ferroptotic cell-based experimental strategies for cancer immunotherapy. In conclusion, these results indicate that induction of ferroptosis in cancer might be another option to overcome cell death resistance and enhance the efficacy of anti-cancer therapy.
Characterization, biodistribution and functional Studies of immune-therapeutic Exosomes: implications for inflammatory lung diseases and COVID 19
Authors: Elashiry Mahmoud , Elsayed Ranya , Cutler Christopher 
Published: February 2023

Abstract
Dendritic cell (DC)-derived exosomes (DC EXO), natural nanoparticles of endosomal origin, are under intense scrutiny in clinical trials for various inflammatory diseases. DC EXO are well-tolerated by the host and can be custom-tailored for immunomodulatory functions. Previously we documented the efficacy of immunoregulatory DCs EXO (regDCs EXO) as immunotherapy for inflammatory bone disease in-vivo. However, the on- and off-target effects of these therapeutic regDC EXO and how target signaling in acceptor cells is activated is unclear. In the present report, therapeutic regDC EXO were analyzed by high throughput proteomics, with non-therapeutic EXO from immature DCs and mature DCs as controls, to identify shared and distinct proteins and potential off-target proteins. The predominant expression in regDC EXO of immunoregulatory proteins as well as proteins involved in trafficking from the circulation to peripheral tissues, prompted us to investigate how these EXO are biodistributed to major organs after intravenous injection. Live animal imaging showed preferential accumulation of regDCs EXO in the lungs. In addition, TGFB1 in regDCs EXO sustained downstream regulatory signaling in acceptor DCs. Blocking experiments suggested that sustaining TGFB1 signaling require initial interaction of regDCs EXO with TGFB1R followed by internalization of regDCs EXO with TGFB1-TGFB1R complex. Finally, these regDCs EXO that contain immunoregulatory cargo and showed biodistribution to lungs could downregulate the main SARS-CoV-2 target receptor, ACE2 on recipient lung parenchymal cells via TGFB1 in-vitro. In conclusion, these results may have important immunotherapeutic implications for lung inflammatory disorders and COVID19.
Designing a bispecific antibody targeting IL-6Rα and NKG2A receptors via in silico affinity maturation as a strategy to combat severe COVID-19
Authors: Agarwal Manusmriti , Saini Samvedna , Kaushik Spandan , Prasad Aakanksha , Kumar Yatender 
Published: February 2023

Abstract
High mortality in COVID-19 patients has been encountered with acute respiratory distress syndrome (ARDS). Cytokine storm has been observed as a reason for this severity in critical cases showcasing high levels of circulating cytokines along with a pronounced decrease in cytotoxic T lymphocytes and Natural Killer Cell populations. The Interleukin-6 amplifier, the activation of IL-6-signal transducer and activator of transcription 3 (STAT3) and NF-κB signaling in non-immune cells has been widely discussed as an initiator of this cytokine release syndrome. The anti-IL-6α receptor antibody tocilizumab has shown success in recovery when administered to critical COVID-19 patients. Additionally, increased release of IL-6 is shown to elicit immune cell disproportionation by upregulating inhibitory NKG2A receptors on NK cell surface, causing blunting of the host’s antiviral response. This study proposes a bispecific immuno-modulatory antibody with one Fab blocking IL-6α-receptor and the other blocking NKG2A receptor to counter hyper-inflammatory cascades and recruit NK cells to the site of viral infection, respectively. Computational site directed mutagenesis was performed to design a library of increased binding affinity mutants of tocilizumab and NKG2A inhibitory monalizumab, followed by docking studies, molecular dynamics simulations, MM-PBSA analysis to study stability of the bispecific assembly in physiologically relevant conditions. This approach utilizing immunomodulation for simultaneous recruitment of antiviral immunity and regulation of hyper-inflammatory host responses may present a potent strategy for combatting critical stage COVID-19 and thereby help in curbing COVID-19 related mortality.
The Role of Aire in the selection of Regulatory T cells in Type 1 Diabetes
Authors: Bridge Jennifer Anne , Balolong Jared , Belk Julia , Dong Shen , Cramer Nathan , Mowery Cody , Crawford Frances , Ye Jimmy , Satpathy Ansuman , Kappler John W , Bluestone Jeffrey A , Anderson Mark S 
Published: February 2023

Abstract
In autoimmune diseases like T1D, it is proposed that failure in central tolerance mechanisms towards pancreatic islet self-antigens leads to the escape of pathogenic clones and loss of critical immunosuppressive antigen-specific Tregs. The importance of Aire in maintaining central tolerance against self-reactive antigens is evidenced by the multiorgan autoimmune disease seen in individuals with mutation the Aire gene. When Aire-expression is specifically ablated in mouse mTECs, there is a dramatic increase in the number of self-reactive T cells that escape negative selection, in addition, a dramatic loss of Tregs. Utilizing novel high-throughput platforms and single-cell DNA barcoding technology has allowed us to assess several thousand CD4+ T-cell clones. The use of FoxP3-GFP reporter mice permits us to identify individual FoxP3+ Tregs that have undergone Aire-dependent selection in the thymus and compare the TCR repertoire to mice that have had Aire specifically ablated from the mTECs. Identification of unique clones and gene signatures within our large datasets has allowed us to start to qualitatively assess the functional role of the identified TCRs within the model of T1D. GLIPH2 clustering using our extensive insulin-specific TCRs have identified a number of motifs shared within our TCR dataset, as well as a unique V-gene usage that may predict TCR affinity. Our proposed study aims to expand our knowledge on TCR repertoire of Aire-dependent FoxP3+ Tregs and the antigen-specificity in T1D, improving our understanding of the respective function of thymic-derived antigen-specific Treg suppression in peripheral tissues associated with autoimmune T1D.
Dysregulated follicular regulatory T cells and antibody responses exacerbate CNS autoimmunity
Authors: Leavenworth Jianmei Wu , Luo Lin , Hu Xianzhen , Dixon Michael L. , Pope Brandon J. , Leavenworth Jonathan D. , Raman Chander , Meador William R. 
Published: February 2023

Abstract
Follicular regulatory T (TFR) cells are essential for the regulation of germinal center (GC) response and humoral self-tolerance. Dysregulated follicular helper T (TFH) cell-GC-antibody (Ab) response secondary to dysfunctional TFR cells is the root of an array of autoimmune disorders. The contribution of TFR cells to the pathogenesis of multiple sclerosis (MS) and murine experimental autoimmune encephalomyelitis (EAE) remains largely unclear. Here we show that mice with a FoxP3-specific ablation of the transcription factor Blimp1 developed severe EAE and failed to recover compared to control mice, reflecting conversion of unstable regulatory T cells (Tregs) into IL-17A/GM-CSF-producing effector T-cells associated with increased TFH-Ab responses, more IgE deposition in the central nervous system (CNS) and inability to regulate CNS CD11b+ myeloid cells. Notably, serum IgE titers were positively correlated with EAE scores, and culture of CNS CD11b+ cells with sera from these EAE mice enhanced their activation, while transfer of Blimp1-deficient TFR cells promoted Ab production, activation of CNS CD11b+ cells and EAE. Moreover, compared to healthy controls, a small group of MS patients had fewer circulating Tregs and TFR cells that expressed reduced levels of Blimp1, associated with elevated serum IgE. These findings suggest that Blimp1 is essential for the maintenance of TFR cells and Ab responses in EAE, and that dysregulated TFR cells and Ab responses promote CNS autoimmunity.
T cell dysfunction in Activated PI3K δ Syndrome explained by cell intrinsic and extrinsic components
Authors: Bier Julia , Lau Anthony , Brink Robert Adrian , Ruiz-Diaz Stephanie , Lenthall Helen , Tangye Stuart , Deenick Elissa 
Published: February 2023

Abstract
Activated PI3K Delta Syndrome (APDS) is a rare condition caused by heterozygous gain of function mutations in PIK3CD, which encodes the leukocyte-restricted p110δ catalytic subunit of phosphoinositol 3-kinase (PI3K). PI3K is activated downstream of many T cell receptors and has been implicated in the control of CD4+ T cell helper differentiation, activation and proliferation. APDS patients have increased PI3K activity that leads to several immune manifestations including lymphoproliferation, Th2-related pathologies and impaired Ab responses.
Analysis of a CRISPR/Cas9 mouse model (Pik3cdE1020K) showed a decreased frequency of naïve and a concurrent increase in memory T cells and Tfh cells, indicating T cell hyperactivation, as well as changes in cytokine expression and T cell help. Our studies investigated CD4+ T cell intrinsic and extrinsic alterations contributing to the dysregulation of the immune response.
We identified cell intrinsic features resulting in increased IL-4 and IL-5, explaining the Th2 mediated disease in these patients. Additionally, cell intrinsic changes in Tfh generation and function were shown in the in vivo mouse model, contributing to an impaired ability to sustain GCs and alter isotype switching. Furthermore, additional studies have shown that the overactive T cells result from both intrinsic and extrinsic factors, with contributions from both dysregulated T cells and B cells exerting effects on T cell activation.
Together, these studies reveals an important role of PI3K in modulating T cell differentiation and function through both T cell-intrinsic and extrinsic mechanisms. This provides new insights into the APDS pathophysiology with potential to improve the clinical management of these patients.
γHV68 infection exacerbates arthritis and requires age-associated B cells
Authors: Mouat Isobel , Morse Zachary J. , Shanina Iryna , Brown Kelly L , Horwitz Marc S 
Published: February 2023

Abstract
Epstein-Barr virus (EBV) infection is associated with rheumatoid arthritis (RA), though the mechanism of contribution remains unknown and there does not exist a sufficient in vivo model to examine the relationship. Here, we utilize and expand in vivo models of EBV and RA to examine mechanisms of immune contribution. We find that infection with latent gamma-herpesvirus 68 (γHV68), a murine analogue of EBV, leads to an enhanced clinical and immunological course of collagen-induced arthritis (CIA). γHV68-infected mice display earlier and more severe CIA clinical symptoms and a Th1-skewed immune profile, compared to uninfected CIA mice. Using a latency-free strain of γHV68 we demonstrate that CIA exacerbation is not due to innate immune stimulation during acute infection but, rather, is dependent upon viral latency. Age-associated B cells (ABCs) are increased in RA patients and during viral infection, though if they act as mediators between the infection and disease remains unknown. We find that ABCs (CD19+CD11c+Tbet+) in γHV68-infected CIA mice are increased and display a proinflammatory phenotype compared to uninfected CIA. Using ABC knockout mice, we demonstrate that ABCs are critical for γHV68-enhancement of CIA, though are dispensable in uninfected CIA. This project establishes that latent γHV68 infection enhances CIA and is a viable model for examining mechanisms of EBV’s contribution to RA. Additionally, we demonstrate that ABCs mediate the viral enhancement of disease.