bluuebunny/crossref_metadata_2025_split huggingface dataset:
Eosinophils alleviate lung allograft rejection through their modulation of CD8+ T Cells
Authors: Onyema Oscar Okwudiri , Guo Yizhan , Wang Qing , Gelman Andrew E. , Kreisel Daniel , Jacobsen Elizabeth A. , Krupnick Alexander Sasha 
Published: January 2023

Abstract

Introduction
Based on the accumulation of eosinophils (eos) in rejected lung grafts, it was widely held that eos perpetuate lung graft rejection. However, we showed recently in a mechanistic study that eos mediate lung graft tolerance. As eos are also present in rejected lungs, we therefore explore the possibility of a different role of eos in lung graft rejection.

Method
Using orthotopic left lung transplantation of MHC-1 mismatched Balb/c (H2Kd) donors to C57BL/6J (H2Kb) recipients, with or without co-stimulatory blockade immunosuppression, we quantified the number of eos in lung grafts at day 7 post-transplant (D7). Th1, Th2 and Th17 cytokine profiles of D4 lung grafts were analyzed by quantitative PCR. D7 eos were also analyzed for their expression of Th1, Th2 and Th17 signature genes. The role of eos was examined in iPHIL mice that experience selective ablation of eos after Diphtheria toxin (DT) administration. Balb/c left lungs transplanted into DT or vehicle treated iPHIL mice without immunosuppression were analyzed histologically and flow cytometrically at D4.

Results
We observed similar numbers of eos, higher Th1 polarization, lower IL33, a Th2 cytokine and less Th17 polarization of the lung microenvironment, and higher expression of the Th1 signature genes among eos, in rejecting compared to accepting lung grafts. Eos deficiency was associated with more rapid lung graft rejection, evidenced in higher CD8+ T cell proliferation, lower CD4/CD8 ratio, and enhanced T cell differentiation to effector memory phenotype.

Conclusion
Our results indicate that eos directly ameliorate lung graft rejection even in the absence of immunosuppression. This calls for a revalidation of the importance of eos in lung allograft pathology.
Type-1 interferon impairs the immunoregulatory activity of IL-10: Understanding the mechanisms of abrogation of transplant tolerance
Authors: Lozano Marcos Iglesias , Arun Anirudh , Chicco Maria , Lam Brandon , Ivanova Vera , Lee W.P. Andrew , Brandacher Gerald , Raimondi Giorgio 
Published: January 2023

Abstract
Long term transplant survival by costimulation blockade (CoB)-based regimens is impaired or eroded by inflammatory responses. Type-1 interferons (TI-IFN) are key mediators of this effect, however, their target population and mechanism of action remain unknown. To better understand how IFN-β interferes with the induction of transplant tolerance, we studied its impact on the recently discovered immunomodulatory properties of interleukin 10 (IL-10) in mouse T cells.
We first demonstrate that IL-10 has an unappreciated fundamental role in transplant tolerance, as its transient blockade in recipients of full mismatch skin grafts impairs the efficacy of a peri-transplant CoB regimen based on donor specific transfusion (DST) plus anti-CD154. Moreover, when T cells are exposed to IFN-β in vitro, they develop a selective dramatic defect in their response to IL-10 (but not to IL-6) both at the signaling and gene-induction level. This IL-10-specific unresponsiveness is not associated with any reduction of IL-10R expression or an increase in SOCS1 and 3, nor with a reduced STAT3 cytoplasmic availability. Instead, our gene-expression and functional analysis suggests a novel “cross-regulatory” role for the transcription factor STAT1. Encouragingly, this inhibition is slowly reversible with the removal of TI-IFN, proving the feasibility a pharmacological intervention. Overall, these results reveal a new molecular mechanism whereby IFN-β interferes with the suppressive function of IL-10 in T cells, and potentially with regulation of alloreactivity. Identifying a strategy to target this mechanism will then be a powerful tool to improve the efficacy of CoB-based strategies for transplant tolerance induction.
Clonal dynamics of adoptively transferred multi-virus specific T cells (MVST) in hematopoietic stem cell transplant patients
Authors: Davies Sarah I. , Yu Quan , Battiwalla Minoo , Chinian Fariba , Whitehill Gregory , Wong Susan , Koklanaris Eleftheria , Hauffe Sara , Stroncek David , Superata Jeanine , Barrett John , Muranski Pawel 
Published: January 2023

Abstract
Hematopoietic stem cell transplant (HCT) patients often risk complications from viral infection due to loss of protective antiviral immunity. One strategy to overcome this hurdle is to supplement the HCT with multi-virus specific T cells (MVST). In phase I trial (NCT02231853), MVSTs were generated using HLA-matched donor-derived dendritic cells pulsed with viral overlapping peptide libraries (cytomegalovirus (CMV), Epstein-Barr virus, Adenovirus, and BK polyomavirus) and then co-cultured with donor lymphocytes. These cells were infused into the patient in early post-SCT. Using T cell receptor CDR3 immnuosequencing, we investigated the fate of adoptively transferred donor derived MVST in four donor-patient pairs. Relative clonal frequencies were analyzed in the peripheral blood and bone marrow day 60, 100, 180, and 1 year post transplant. The top 10 most frequent clones in the MVST product did not necessarily survive in the patients, and only minor subsets of MVST clonal sequences survived at day 100 (2.92–7.75%). However, these minor MVST clones quickly expanded in all four patients to constitute over &gt;20% of peripheral blood T cells even 6 months to 1 year later. Two patients analyzed had MVST antiviral clones peak during and after detectable viremia or viriuria, but were asymptomatic. One patient received steroids for treatment of GVHD, which was accompanied by the decline of MVST clones and subsequent CMV reactivation. These results suggest that adoptive transfer of viral-specific T cells can promote reconstitution of the anti-viral T cell repertoire and these cells persist in patients long-term post transplant. Furthermore, the fittest clones are dynamically selected to serve as long term sentinels against viral reactivation.
Role of exosomes in rejection following human lung transplantation – an in vitro analysis using human airway epithelial cells
Authors: Ravichandran Ranjithkumar , Gunasekaran Muthukumar , Bansal Sandhya , Mohanakumar Thalachallour 
Published: January 2023

Abstract
Studies have demonstrated that exosomes expressing donor human leukocyte antigen (HLA) and lung self-antigens (SAg), Collagen V (Col-V) and Kα1 Tubulin (Kα1T), can be detected in the circulation of human lung transplant recipients (LTxR) who developed antibodies (Ab) to donor HLA and diagnosed with rejection However, the mechanisms by which Ab mediate induction of exosomes into the circulation from airway epithelial cells (AEC) remains unclear. Ab ligation to HLA could enhance PERK-mediated pathways involved in endoplasmic reticular (ER) stress-dependent exosome release. The goal of this study is to determine the mechanism by which Ab induce exosomes from AEC. Towards this we employed human AEC, ligated antigens by Ab specific to HLA (HiPRA) and lung SAg (rabbit anti-Col-V or Kα1T) and the role of ER stress signaling molecule in exosomes release were determined. Effect of exosome inhibition by GW4869 was also analyzed. AEC were incubated with HiPRA or Ab to SAg for 24hr in both normoxia and hypoxia (0.1% O2) conditions. AEC incubated with HiPRA or Ab to SAg demonstrated induction of exosomes containing lung SAg, Col-V and Kα1T. Incubation with HiPRA and Ab (20μg/ml) to SAg also demonstrated induction of exosomes containing lung SAg, MHC class II, CD80, CD86. Treatment with GW4869, exosome inhibitor (20μM) reduced the exosomes and SAg. Effect of HiPRA and Ab to SAg on stress kinase in both normoxic and hypoxic conditions and influence of GW4869 in these pathways will be delineated. Identification of stress kinase and exosome inhibition will provide novel results for Ab mediated upregulation of stress leading to exosome release from the transplanted organ resulting in immune responses thus increasing the risk for chronic rejection following LTx.
Identification of extracellular vesicle proteins in circulating exosomes from human lung transplant recipients
Authors: Bansal Sandhya , Pirrotte Patrick , McGilvrey Marissa , Garcia-Mansfield Krystine , Sharma Monal , Gunasekaran Muthukumar , Smith Michael , Bremner Ross , Mohanakumar Thalachallour 
Published: January 2023

Abstract
Molecular mechanisms involved in rejection following human lung transplantation (LTx) are not well understood. We identified proteomic signatures of clinical outcomes in circulating extracellular vesicles (EVs) isolated from human lung transplant recipients (LTxRs) who was diagnosed with chronic rejection (Bronchiolitis Obliterans Syndrome (BOS)), acute rejection (AR), respiratory viral infection requiring intervention (RVI) or stable following transplantation. Differential analysis revealed one protein unique to AR (Skin-specific protein 32), four unique to RVI (Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2, Transmembrane 9 superfamily member 2, Ras-related C3 botulinum toxin substrate 2, and EH domain-containing protein) and two unique to stable (Coagulation factor X, and N-acetylmuramoyl-L-alanine amidase). Comparison of each rejection group to stable identified 128, 180, and 216 significantly differentially expressed proteins (p-value&lt;0.05, fold-change&gt;2) in BOS, AR, and RVI respectively. Although no unique signatures were found in BOS, an increased enrichment of immune processes such as antigen processing and presentation of exogenous peptide antigen via MHC class I and Fc receptor signaling pathway were observed. Functional enrichment analysis (Gene Ontology) associated differentially expressed EV proteins in AR and RVI conditions with wound repair processes. These diverse signatures detected in each condition highlight complex immune mechanisms underlying the pathophysiology of rejection following LTx. Future studies are needed to validate these signatures in larger LTxR cohorts in order to promote early clinical diagnosis and predict outcome.
Urine levels of CCL2 and CXCL10 chemokines as potential biomarkers of kidney allograft function: a pilot study.
Authors: Czerwinska Magdalena , Gniewkiewicz Michał , Gozdowska Jolanta , Czerwińska Katarzyna , Sadowska Anna , Perkowska-Ptasińska Agnieszka , Kosieradzki Maciej , Durlik Magdalena 
Published: January 2023

Abstract

Introduction
Early prognostic markers that identify high-risk patients could lead to increased surveillance, personalized immunosuppression, and improved outcomes. The vigilance and rapid detection of allograft injury is what would result in the identification of patients who will require more intensive monitoring to ensure that their new allografts function well for the longest possible time.
The aim of the study was to assess whether urine levels of CCL2 and CXCL10 can be valuable, noninvasive source of information about inflammation, tubular injury, progression of fibrosis and other damage features in the kidney allograft.

Materials and methods
In this prospective study 40 patients who underwent a protocolar biopsy within 1 year post kidney transplant were included. The levels of chemokines CCL2 and CXCL10 with reference to creatinine in urine were assayed in all patients. Based on biopsy results research group (N=25) with diagnosis of BKV nephropathy, IFTA II–III, peritubular capillaritisis, C4d or rejection was selected. Patients with normal biopsy results were included as control group (N=15).

Results
Median urinary CCL2-to-creatinine (Cr) ratio [ng/mmol] was significantly elevated in research group compared to control group (21.76 ± 18.57 vs 8.99 ±5.12, p=0.003). There was no statistical difference between groups regarding CXCL10/Cr ratio (7.96 ± 9.08 vs 5.58 ±5.7 p=0.32). Patients with BKV nephropathy (N=3) had much higher level of CCL2 [ng/l] than control group but the difference did not reach statistical significance (429.69 ± 166.49 vs 78.04 ±53.61, p=0.06).

Conclusion
CCL2/Cr may be used as a non-ivasive tool useful for monitoring graft condition and predicting recipients with high risk of transplant kidney loss.
Towards the repair of marginal liver grafts: Mesenchymal stromal cell therapy.
Authors: Bartczak Agata , Linares Ivan , Kollmann Dagmar , Zhang Xing-Hua , Ma Xue-Zhong , Echeverri Juan , Ganesh Sujani , Manuel Justin , Selzner Markus , Keating Armand , McGilvray Ian 
Published: January 2023

Abstract
Despite the recent inclusion of livers donated after cardiac death (DCD), more than 25% of people listed for a liver transplant in Canada die before obtaining a suitable graft. Over half of possible DCD livers undergo long periods of ischemia that sensitize the liver to reperfusion injury (HIRI) and render the liver grafts unsuitable for transplantation. Mesenchymal Stromal cells (MSC) show promise in protecting liver tissue against HIRI: MSC stimulated with inflammatory agents have enhanced anti-inflammatory activity. Kupffer Cells (KCs) are central to HIRI, are activated after reperfusion, and release various inflammatory mediators that promote hepatocyte death. We hypothesize that LPS-primed MSC-treatment can mitigate the tissue damage inflicted by HIRI by modulating KC activation. To test this, we injected LPS-primed MSCs(LPS) into rats undergoing warm ischemia to 70% of the liver. Rats that received MSCs(LPS) had significantly lower levels of AST (2288 ± 2121 IU/L) compared to sham controls (6614 ± 3495 IU/L). Histological analysis showed decreased TUNEL and Caspase-3 positive staining in the MSC(LPS) group: the necrotic area in the MSC- and sham-treated groups affected &gt;50% of the liver parenchyma whereas, only ~20% was necrotic in the MSC(LPS) group. In this latter group, the number of infiltrating inflammatory CD68+ macrophages in the necrotic tissue and CD163+ macrophages in the viable tissue was significantly decreased. Systemic treatment of rats with activated MSCs resulted in the decreased migration of macrophages into inflamed tissue and limited parenchymal tissue damage. Primed MSCs are a viable treatment to protect marginal DCD donor liver grafts from the damaging effects of HIRI.
The adapter protein TSAd maintains alloimmune T regulatory cell stability and function
Authors: Wedel Johannes , Stack Maria P. , Seto Tatsuichiro , Sheehan Matthew M. , Liu Kaifeng , Flynn Evelyn , Briscoe David M. 
Published: January 2023

Abstract
TSAd is an SH2 domain containing intracellular adapter molecule that binds to Lck and functions to elicit T cell receptor-mediated signals and cytokine production. Deficient expression has been associated with autoimmunity in humans and mice, but its mechanism of function in Tregs is not known. We used TSAd KO mice to evaluate its function in CD4+ Tregs, and find that KO Tregs suppress effector proliferation in vitro similar to WT Tregs. However, TSAd KO Tregs displayed higher methylation of the TSDR region of the Foxp3 gene suggesting that TSAd may function via epigenetic regulation of Treg stability. TSAd KO Tregs also produced significantly more effector IFNγ upon mitogen stimulation (vs. WT) indicating that they possess proinflammatory function. To assess function in vivo, we transplanted MHC class II mismatched B6.C-H-2bm12 heart transplants into C57BL/6 WT or TSAd KO recipients. While allograft survival was &gt;50 days in WT recipients, allograft rejection was markedly accelerated in TSAd KO recipients (mean survival time [MST] 22 days, P&lt;0.01). By FACS, CD4+CD44hiCD62Llo Teffs expanded in TSAd KO recipients (vs. WT) supporting the possibility that TSAd KO Tregs are less efficient in suppression and/or possess effector function. To further evaluate these possibilities, we adoptively transferred CD4+CD25+ WT or TSAd KO Tregs into C57BL/6 Rag2γc KO recipients of fully MHC mismatched Balb/c allografts on day 2 post-transplant; subsequently WT CD4+CD25− Teffs were transferred on day 18. In this model WT Tregs (MST 68 days) were significantly more efficient than TSAd KO Tregs (MST 48, P&lt;0.05) in suppression of effectors in vivo. Thus, TSAd provides cell-intrinsic signals to suppress proinflammatory effector function in Treg populations.
A novel function of platelets and CD40L in NAFLD-promoted HCC development
Authors: Fu Qiong , Ma Chi , Heinrich Bernd , Brown Zachary , Zhang Qianfei , Elzey Bennett , Ratliff Timothy , Greten Tim 
Published: January 2023

Abstract
Hepatocellular carcinoma (HCC) is the 2nd leading cause of cancer-related death worldwide. Chronic liver inflammation, such as non-alcoholic fatty liver disease (NAFLD), which is characterized by increased fat deposition in the liver, is one common cause of HCC. The prevalence of NAFLD is increasing rapidly with the growing epidemics of diabetes and obesity, but little is known about how NAFLD proceeds to HCC.
Platelets, which are a critical component in hemostasis, have been shown to release CD40L after activation, and there is accumulating evidence for an important role of platelets in adaptive immunity. Platelet-derived CD40L, by interacting with its receptor CD40, is able to induce dendritic cell maturation, prime T cells, and stimulate B cell isotype switching.
NAFLD patients have increased mean platelet volume, which is an indicator of platelet activation, and an increased level of CD40L in their serum. However, this potential role of platelets in NAFLD-promoted HCC is completely unknown.
Here we studied the platelet function in a murine HCC model of NAFLD and HCC.
We found that inhibition of platelet activation using Aspirin and Clopidogrel greatly increased HCC incidence. Similar results were found when mice were treated with a CD40L blocking antibody, while the combination treatment did not show an additive effect. However, Aspirin and Clopidogrel treatment had no effect on HCC in the absence of NAFLD, while using only a CD40 activating antibody significantly reduced HCC incidence without NAFLD. Finally, an increased CD40L level was found in the serum from mice with NAFLD using a bioassay. In summary, our results suggest an important role of platelets in NAFLD-promoted HCC, and will provide useful information for HCC treatment.
Systemic dysregulation of antigen cross-presenting dendritic cells occurs early in preinvasive pancreatic neoplasia and is reversed by CD40 agonism
Authors: Lin Jeffrey H. , Vonderheide Robert H. 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDAC) ranks as the third-leading cause of cancer-related deaths in the United States with a five-year survival rate of 9%. While immune checkpoint blockade (ICB) has revolutionized treatment of many metastatic diseases, cancers like PDAC that lack tumor-infiltrating CD8+ T-cells remain refractory to ICB. To determine whether this phenotype is due to defective CD8+ T-cell priming by antigen cross-presenting XCR1hi classical dendritic cells (cDC’s), we use the KPC mouse model of PDAC driven by Pdx1-Cre KrasLSL-G12D/+Trp53LSL-R172H/+ to interrogate how XCR1hi cDC’s change over the course of cancer development. In the present study, flow cytometric analysis reveals a progressive systemic decline in XCR1hi cDC differentiation and activation that begins in mice bearing preinvasive pancreatic intraepithelial neoplasias (PanIN’s). These changes occur in the setting of intact cDC maturation, consistent with induction of a tolerogenic program, and strengthen upon progression to malignancy. Administration of CD40 agonist to tumor-bearing mice effectively reverses defects in XCR1hi cDC activation and drives CD8+ T-cell infiltration into the tumor microenvironment when combined with the chemotherapies gemcitabine and nab-paclitaxel. Our results therefore demonstrate that antigen cross-presenting cDC’s are dysregulated early and prominently in the neoplastic process and position CD40 agonism as an ideal strategy to potentiate ICB in PDAC.
The coupling of MDSCs with a computational neural network (NN) to detect solid tumors.
Authors: Dominguez George A. , Maslar Kristen , Sholevar Cyrus , Polo Alexander , Roop John , Campisi Anthony , Gabrilovich Dmitry I. , Rauscher Frank J. , Kumar Amit 
Published: January 2023

Abstract
The goal of this study was to create a non-invasive cancer detection assay that analyzes flow cytometry data in an objective method. By using an artificial neural network (NN), we were able to distinguish between cancer patients (CPs) and healthy donors (HDs) based upon the flow cytometry profiles of MDSCs and other leukocytes. Myeloid-derived suppressor cells (MDSCs) are known to be key contributors in supporting tumor progression and tumor escape through their ability to suppress anti-tumor responses. Several studies have demonstrated their utility as indicators for tumor development, but there is significant overlap with healthy individuals. We used standard multiparametric flow cytometry techniques to immunophenotype MDSCs and other leukocytes found in the peripheral blood of 65 biopsy-confirmed CPs with solid tumors and 84 HDs. A series of NNs utilizing pattern recognition computational algorithms were created using three data sets: 1) the training set – this ‘teaches’ the two output categories of cancer and not cancer, 2) the validation set – this uses backpropagation to improve the accuracy of the trained networks, and 3) the testing set – this is used to rank the trained networks against each other. Finally, a naïve testing set is then used to determine the overall sensitivity and specificity for the top-ranking networks. Here, we incorporated a standard 12 marker flow cytometry assay with NN technology to achieve a sensitivity of 92% and a specificity of 89%. Pairing the advanced analytical capabilities of our NN with surface biomarker based analysis of MDSCs and certain leukocytes measured in peripheral blood has enabled us the ability to objectively identify patterns indicative for the existence of a solid tumor.
Relevance of host and tumor PD-L1 in PD-L1 pathway blockade
Authors: Kryczek Ilona , Lin Heng , Wei Shuang , Green Michael , Zou Weiping 
Published: January 2023

Abstract
PD-L1 and PD-1 pathway blockade is a promising therapy against cancer. However, the mechanistic contribution of host and tumor PD-L1 and PD-1 signaling to therapeutic efficacy of PD-L1 and PD-1 blockade remains elusive. Using three tumor bearing mouse models that differ in their sensitivity to PD-L1 blockade, MC38, ID8, and B16-F10, we found a loss of therapeutic efficacy of PD-L1 blockade in immunodeficient mice and in PD-L1 and PD-1 deficient mice. In contrast, knockout or overexpression of PD-L1 in tumor cells had no effect on PD-L1 blockade. Human and murine studies show high levels of functional PD-L1 expression in dendritic cells and macrophages in the tumor microenvironments and draining lymph nodes. Further, expression of PD-L1 on dendritic cells and macrophages in ovarian cancer and melanoma patients correlates with the efficacy of anti-PD-1 and anti-PD-1 plus anti-CTLA-4 therapy. Thus, PD-L1+ dendritic cells and macrophages may mechanistically shape and therapeutically predict clinical efficacy of PD-L1/PD-1 blockade.