bluuebunny/crossref_metadata_2025_split huggingface dataset:
Repression of NFAT1 protein expression by MicroRNA-184 in umbilical cord blood (UCB) CD4+ T-cells as a potential mechanism underlying lower incidence of acute graft-vs-host disease (aGVHD) (145.36)
Authors: Weitzel R. , Lesniewski Mathew , Laughlin Mary 
Published: January 2023

Abstract
Lower expression of the transcription factor NFAT1 in UCB CD4+ T-cells compared to adult blood (AB) results in reduced inflammatory cytokine production after stimulation, in part underlying its advantages in unrelated transplant. Fractionation and RT-PCR reveal that NFAT1 is translationally repressed in UCB during early stimulation, which may be due to microRNA. microRNA queries reveal that miR-184 is strongly predicted to bind to the NFATc2 3’ untranslated region (UTR). In UCB, insertion of the miR-184 binding site into the luciferase 3’ UTR reduced expression by 62% and the full 3’ UTR by 40%. Both these effects were reversible by cotransfection with excess of blocking antisense to miR-184. The UTR did not impact luciferase expression in AB CD4+ T-cells, but both luciferase expression and NFAT1 protein were reduced in AB by a miR-184 mimic in separate analyses, also reversible by cotransfection of antisense. This antisense increased NFAT1 protein expression in UCB CD4+ T-cells as measured by Western Blot without reducing NFATc2 mRNA. qRT-PCR confirmed that miR-184 is expressed 58.4-times greater in UCB than in AB CD4+ T-cells, possibly due to methyl-CpG DNA-binding proteins. We also demonstrated that the effects of miR-184 on NFAT1 are sufficient to alter transcription of the NFAT-dependent cytokine IL-2. This work may hold implications for understanding GVHD, neonatal lymphocyte activation, and suggest miR-184 as a useful biomarker or target of ex vivo graft analysis.
Cyclin-dependent kinase 2 (CDK2) opposes T cell anergy and promotes cardiac allograft rejection (145.41)
Authors: Chunder Neelanjana , Wang Liqing , Hancock Wayne , Wells Andrew 
Published: January 2023

Abstract
T cell differentiation and anergy are linked to the cell cycle, but the molecular basis of this relationship is not understood. Cyclin-dependent kinases (CDK) represent a major class of cell cycle proteins that could potentially regulate T cell function. Using a combination of dominant-negative, shRNA knock-down, and genetic deletion approaches, we show that the TCR/CD28 complex is specifically coupled to the il2 gene through CDK2. Additionally, a chemical genetic approach using CD4+ T cells expressing an analog-sensitive mutant of CDK2 resulted in dose-dependent inhibition of IL-2 by a specific analog inhibitor. Importantly, transient inhibition of CDK2 activity resulted in a stable state of anergy. T cells lacking CDK2 exhibited defective in vivo alloimmune responses, and mice deficient for CDK2 failed to reject cardiac allografts under conditions of limiting CD28 costimulation. Conversely, deletion of the CDK inhibitory protein p27kip1 resulted in resistance to T cell anergy and failed induction of transplantation tolerance in response to costimulatory blockade. These results suggest that CDK2 is a component of a direct signaling pathway from TCR/CD28 to the il2 gene that is required for anergy avoidance, and implicate CDK2 as a potential target for immunosuppressive therapies that may promote immunological tolerance.
Alloreactive CD8 T cells require cell-intrinsic PD-1 and upregulate Nr4a and Egr family members during deletional tolerance in vivo (145.43)
Authors: Lucas Carrie , Sykes Megan 
Published: January 2023

Abstract
Objective: To characterize deletional tolerance of peripheral alloreactive CD8 T cells induced by anti-CD40L and allogeneic BMT. Methods: B6 mice received 3Gy TBI on Day -1 and 2mg anti-CD40L ip prior to B10.A BMT on Day 0. For CD8 transfer, KbDb KO mice were given 6-9x10^6 purified, recipient-type CD8 cells on Day 0. For deletion and microarray studies, synchimeras containing 10-20% 2C TCR Tg (donor-reactive) cells among CD8 T cells were used. Results: Using adoptive transfer, we determined that the inhibitory PD-1 receptor is required on CD8 T cells for tolerance (6/7 vs. 0/8 chimeras with WT vs. KO CD8s). 2C cells in animals treated with blocking anti-PD-1 initially disappeared at a similar rate to those in tolerized mice. On Day 6 the frequency of apoptotic 2C cells in tolerant controls was equivalent to that in anti-PD-1-treated mice (p = 0.2) and greater than that in mice with no mAb (p = 0.03). Following complete rejection of donor BM, 2C cells in anti-PD-1-treated mice expanded (p = 0.004 vs. tolerant at Day 42). Microarray analysis on sorted 2C+ CD8 T cells isolated on Day 6 post-BMT revealed significantly higher transcript levels for the apoptosis-associated Nr4a family members Nur77 (7.3 fold, p = 0.01), Nurr1 (23 fold, p = 0.01), and Nor1 (9 fold, p = 0.007) and the tolerance-associated Egr family members Egr-1 (4 fold, p = 0.01), Egr-2 (15 fold, p = 0.001), and Egr-3 (8 fold, p = 0.04) in tolerant compared to rejecting mice.
Determining the extent to which clinically effective treatment, ustekinumab or etanercept, reverses the molecular disease profile of psoriatic skin: comparisons of lesional, non-lesional and normal skin(34.5)
Authors: Brodmerkel Carrie , Li Katherine , Baribaud Frédéric , Suarez-Farinas Mayte , Krueger James 
Published: January 2023

Abstract
ACCEPT, a randomized, active-controlled study, compared the efficacy of etanercept and ustekinumab in 903 moderate-severe plaque PsO pts through wk12. Skin bx were performed in a subset of pts at baseline(BL), wk1 & wk12. Microarray analyses(Affymetrix U133+2 array) comparing nonlesional skin(NL,n=85) to lesional skin(n=85) at BL showed several thousand probe sets differentially expressed(&gt;2fold change FDR,p&lt;0.05) in lesional skin. 25 healthy skin bx were analyzed. Comparison of NL to healthy skin showed a series of lesional genes dysregulated in NL(DEFB4,S100A7A,CCL18,SERPINB3). Analyses to understand the impact of p40 cytokine(IL12/23)or TNFα blockade on resident and inflammatory cells and on expression of gene circuits that may drive chronic immune activation and inflammation in skin were done. Analyses to understand residual molecular profile or “molecular scar” following 12wk of tx were completed. Pts responding to each agent(≥PASI75,ETA n=21,UST n=19)had signif changes in ∼4000 transcripts compared to untreated lesions, indicating signif resolution of pathological gene circuits back to NL levels. However, genes such as DEFB4, S100A7, CCL18 & SERPINB3 though reduced to levels similar to NL were not reduced to that of healthy skin by either tx unlike ADAM10 & HSD3B1. Elucidation of molecular pathways which remain dysregulated following effective tx may provide insight into pathological mechanisms that remain active despite appearance of clinical and histologic resolution.
Th17 cells contribute to the exacerbated autoimmune arthritis in CCR2-deficient mice (34.6)
Authors: Liu Peng , Tarrant Teresa , Rampersad Rishi , Vallanat Christopher , Quintero-Matthews Tatiana , Patel Dhavalkumar , Fong Alan 
Published: January 2023

Abstract
Th17 cells have been implicated in the pathogenesis of autoimmune diseases including rheumatoid arthritis (RA). Chemokine receptor CCR2 mediates leukocyte trafficking to sites of inflammation and is considered to be an important proinflammatory factor in RA. However, as an unexpected paradox, mice lacking CCR2 (CCR2-/-) develop an accelerated and enhanced arthritis in the collagen induced arthritis model (CIA). To investigate the underlying mechanism(s), we examined the role of Th17 cells in CCR2-/- mice with CIA. We found that the number of Th17 cells was increased significantly in draining lymph nodes of CCR2-/- CIA mice (845±144) compared to WT controls (289±136, p=0.017). In contrast, Th1 cells were not significantly different between groups (485±171 vs. 516±86 for WT, p=0.88). Consistently, CCR2-/- CIA mice had elevated levels of sera IL-17 (84.4±15.5 pg/ml) and IL-17 mRNA in arthritic joints (3.43±0.61 fold) compared to WT mice (45±9.7 pg/ml in the serum and 0.21±0.05 fold in the joint, p=0.04 and 0.009 respectively). Furthermore, Th17-favored cytokines IL-6 and IL-1β were up-regulated in the serum of CCR2-/- animals (IL-6: 366.1±54.5 pg/ml vs. 58.7±13.3 pg/ml for WT, p=0.0004; IL-1β: 189.6±21.9 pg/ml vs. 106.4±21.3 pg/ml for WT, p=0.01). These data suggest that Th17 cells contribute to the pathogenesis of the aggravated phenotype of autoimmune arthritis in CCR2-/- mice. Mice deficient in CCR2 may serve as a model for evaluations of anti-Th17 therapies against RA.
Regulating human Th17 cells via differential expression of IL-1 receptor (34.7)
Authors: Lee Won-Woo , Kang Seong Wook , Choi Jihoon , Lee Seung-Hyun , Shah Kamini , Eynon Elizabeth , Flavell Richard , Kang Insoo 
Published: January 2023

Abstract
In humans, IL-1β is essential for developing Th17 cells. However, little is known about the relationship of IL-1 receptor (IL-1R) expression and Th17 cell differentiation. We report the presence of two distinct CD4+ T cell populations with and without expression of IL-1RI that correlates with the capacity to produce IL-17 in human naive and memory CD4+ T cells. A combination of T cell receptor (TCR) triggering and IL-1β augmented IL-17 production from IL-1RI+ memory CD4+ T cells. IL-1RI+ memory CD4+ T cells in peripheral blood had increased gene expression of IL17, RORC and IRF4 even before TCR triggering, suggesting that the programmed effect of IL-1β in these cells. Of interest, IL-1β also promoted IL-17 production from TCR-stimulated IL-1RI- memory CD4+ T cells, which was likely secondary to TCR-mediated up-regulation of IL-1RI. Adding IL-1β to TCR triggering resulted in production of IL-17 from IL-1RI+ but not from IL-1RI- naive CD4+ T cells. While CD4+ T cells from umbilical cord blood did not express IL-1RI, the cytokines IL-7, IL-15 and TGF-β up-regulated IL-1RI expression on naive CD4+ T cells, suggesting that IL-1RI+ naive CD4+ T cells develop in periphery. Furthermore, IL-17 production from the cytokine-treated IL-1R1- naive CD4+ T cells was induced by IL-1β and this induction was blocked by IL-1R antagonist. These results indicate that human Th17 cell differentiation is regulated via differential expression of IL-1RI which is controlled by IL-7 and IL-15.
IFNβ: an anti-inflammatory cytokine which inhibits dendritic cell migration and proinflammatory cytokine production (35.1)
Authors: Yen Jui-Hung , Kong Weimin , Ganea Doina 
Published: January 2023

Abstract
IFNβ is an approved therapeutic option for the treatment of autoimmune disease, MS/EAE. However, the molecular mechanisms underlying the effects of IFNβ in MS/EAE are not fully understood. In EAE, migrating CD4+ T cells and dendritic cells (DC) are essential for development of clinical symptoms. Expressions of CCR7 and MMP-9 are required for DC migration from the inflammatory site to draining lymph nodes for antigen presentation and activation of naïve T cells and to the CNS during chronic neuroinflammation. Our results show IFNβ inhibits CCR7 expression, MMP-9 production, and migration of DCs matured with different stimuli, including LPS, Poly I:C, TNFα+IFNα, or TNFα+IL-1β+IL-6 in the presence of PGE2. STAT-1 activation is required for the IFNβ-mediated inhibition of CCR7 and MMP-9 expression, and subsequent DC migration. In addition, IFNβ inhibits the production of the proinflammatory cytokines IL-12 and IL-23, whereas promoting the expression of the antinflammatory cytokine IL-10. In vivo administration of IFNβ results in doneregulation of CCR7 and MMP-9 expression, while upregulation of IL-10 production in splenic CD11c+ DC. There is reduced in vivo migration of exogenous DC treated with IFNβ, and of endogenous DC exposed to FITC in mice inoculated with IFNβ. In conclusion, our results provide novel molecular mechanisms for the beneficial effects of IFNβ in MS/EAE through the inhibition of mature DC migration and proinflammtory cytokine production.
Targeting the function of extracellular cyclophilins as a novel approach for the treatment of Rheumatoid Arthritis (RA). (35.13)
Authors: Jurjus Rosalyn , Gloor Kayleen , Herzog Dallen , Berman Dana , Constant Stephanie 
Published: January 2023

Abstract
Elevated levels of extracellular cyclophilins A and B are present in the synovial fluid of RA patients and have been reported to correlate with disease severity. Based on the findings that cyclophilins have potent chemotactic properties for various subsets of pro-inflammatory leukocytes, we propose that extracellular sources of cyclophilins may contribute to the development of RA pathology by promoting leukocyte recruitment into synovial spaces and tissues. Using the collagen-induced arthritis (CIA) mouse model of RA, we tested the impact of treating CIA mice with non immunosuppressive analogs of cyclosporine A (CsA) to inhibit the function of cyclophilins. Our findings demonstrate that treatment with non-immunosuppressive CsA leads to a significant reduction in CIA clinical disease severity, with corresponding reductions in histopathology and levels of pro-inflammatory cytokines and enzymes within the joints. Inhibiting the function of cyclophilin A might provide a novel approach for reducing RA-mediated joint inflammation.
Didox (Dihydroxybenzohydroxamic acid) ameliorates macrophage AhR activation, CYP1A1/AhR Induction, and oxidative stress profiles induced by PCB-126 (35.16)
Authors: Rice Charles , Gallichio Vincent , Matsebatlela Thabe 
Published: January 2023

Abstract
Exposure to AhR-activating xenobiotics is associated with oxidative stress and inflammation. For example, PCB-126 is a potent and well-studied AhR-ligand and inducer of CYP1A1/2 and Phase II metabolic enzymes. Intracellular ROI increases in cells within a few hrs of exposure, long before mRNA transcription and protein induction. Thereafter, levels of ROI continually rise for several days due to a high affinity of PCB-126 for the AhR and recalcitrant metabolism. After a 24 hr exposure of RAW264.7 murine macrophages to 1 µM PCB-126 we show (using qRT PCR) that expression levels of GSH-peroxidase and SOD genes are suppressed, while expression levels of of NADPH-oxidase genes are elevated. As expected, CYP1A1 gene/protein expression is markedly induced and associated with elevated superoxide anion and H2O2 production. In search of strong, but non-toxic antioxidants that may ameliorate AhR-associated ROI and inflammation, we found that Didox (3,4-Dihydroxybenzohydroxamic acid) is very potent. Several GSH-peroxidase genes and GSH reductase, peroxiredoxins, thioredoxin reductase, all three types of SODs, and catalase are elevated by Didox. Furthermore, Didox reverses oxidative stress gene profiles initiated by PCB-126. Of particular note, Didox mimics LPS in that exposure suppresses the expression of both CYP1A1 and AhR genes. We postulate that Didox may have significant potential in ameliorating the toxicity of several classes of AhR-activating immunotoxic compounds.
Blimp1 is not required for all Ig secretion and J-chain expression in nurse shark plasma cells (36.11)
Authors: Doremus Caitlin , Dooley Helen , Ohta Yuko , Flajnik Martin 
Published: January 2023

Abstract
The J-Chain is involved in multimerization of IgM. The oldest animals with Ig isotypes, the cartilaginous fish, express IgM in two forms: a pentameric (19S) form, attached to J-chain, and a monomeric (7S) form. By in situ hybridization adult nurse sharks have splenic IgM-producing plasma cells that are either J-chain positive, and presumed to secrete pentameric IgM, or are J-chain negative 7S secretors. Neonatal nurse sharks produce pentameric IgM, and all plasma cells express J-chain. In addition, some neonatal shark plasma cells, found in discrete splenic locations, express a germline joined isotype, IgM1gj. We hypothesize that during B cell development either 19S producers “switch” to become 7S producers by shutting down J-chain expression, or the two populations develop from separate lineages, with the 7S, J-chain-negative lineage arising later in development than the 19S and IgM1gj lineages. Identification of factors controlling J-chain expression, in shark B cells is necessary to clarify which B cell development pathway is correct. In mammals, Blimp-1 is required for development and maintenance of J-chain expression and immunoglobulin secretion in plasma cells. However, in situ hybridization data show that Blimp-1 is not expressed in neonatal IgM secreting cells, or in half of adult shark secretory cells, presumably the 19S producers. Our data imply that J-chain expression and antibody secretion functions of some shark plasma cells are not controlled by BLIMP1.
Interleukin-2 Controls flt3L-dependent Development and Phenotype of Conventional and Plasmacytoid Dendritic Cells (36.19)
Authors: Lau-Kilby Annie , Kretz Cosima , O'Shea John , Trinchieri Giorgio , Tarbell Kristin 
Published: January 2023

Abstract
DC development requires the ligand for FMS-like tyrosine kinase 3 receptor (flt3L), but little is known regarding other cytokines that may control this process. In our study, we have addressed the effect of IL-2 on DC development. We have shown that the addition of IL-2 to flt3L-stimulated bone marrow (BM) cultures can block the development of both pDCs (CD11c+SiglecH+) and cDCs (CD11c+CD11b+), while expanding the populations of NK/NKT/T cells (CD3+/-NK1.l+/-). By sorting for DC precursors that are Lineage- flt3+ CD11c-, we confirm that IL-2 blocks DC development by acting directly on DC precursors, and not indirectly via its effect on NK/NKT/T cells. The inhibition of DC development by IL-2 is dependent on both the IL-2R alpha and beta chains, and STAT5. IL-2 treatment of DC precursors leads to a decrease in the expression of various transcription factors crucial for DC development. In addition, our ongoing studies indicate that the presence of IL-2 during flt3L-dependent DC development also alters the phenotype of DCs. Together our data has indicated that IL-2 inhibits the development of DCs, and suggests IL-2 can also alter DC phenotype. This has important implications in the therapeutic use of IL-2, as well as our current understanding on factors that can lead to the loss of immune tolerance in various autoimmune diseases.
Cell lineage analysis of KIT D816V and NRAS G12D activating mutations in aggressive systemic mastocytosis (36.25)
Authors: Bai Yun , Simakova Olga , Fu Weiming , Woelfel Jessica , Maric Dragan , Maric Irina , Metcalfe Dean , Wilson Todd 
Published: January 2023

Abstract
Systemic mastocytosis is a clonal disorder characterized by the abnormal proliferation and accumulation of mast cells within tissues. Over 90% of patients with mastocytosis possess a somatic gain of function mutation involving the tyrosine kinase domain of c-kit (KIT D816V). Recently we identified a patient with aggressive systemic mastocytosis who also harbors a somatic NRAS G12D activating mutation. To better understand the clonal evolution of mastocytosis, we evaluated the cell compartments impacted by these two mutations. Bone marrow and peripheral blood cells were collected from the patient. CD34+ progenitors, mast cells, neutrophils, monocytes, eosinophils, basophils, T-lymphocytes and B-lymphocytes were separated by fluorescence-activated cell-sorting. cDNA was prepared and the KIT D816V mutation was tested by nested RT-PCR/RFLP. The NRAS G12D mutation was detected by nested RT-PCR followed by sequencing. The KIT D816V and NRAS G12D mutations were detected in both myeloid and lymphoid lineages. Mast cells harbored both mutations. The KIT D816V mutation was not detected in CD34+ progenitors, whereas presence of the NRAS G12D was observed. These findings suggest that the NRAS G12D mutation occurred prior to the KIT D816V mutation in clonal development. Unlike other mature lineages, mast cell survival is dependent on KIT and the presence of these two mutations may have a greater impact on the expansion of this cell compartment and in resultant disease severity.
Extreme GATA-3 dose dependence in early T cell development (36.28)
Authors: Scripture-Adams Deirdre , Arias Alexandra , Elihu Koorosh , Ho I-Cheng , Rothenberg Ellen 
Published: January 2023

Abstract
GATA-3 expression first reaches peak levels during T-lineage commitment, but a specific requirement for GATA-3 at each of the early DN stages in T development has not been proven. Using retroviral expression of shRNA to reduce GATA-3 protein at each of the earliest stages of T-cell differentiation (DN1, DN2, DN3), we identify two dose-dependent effects: a defect in survival or expansion at DN1, and a separable developmental arrest of cells at DN2, which was cell-autonomous, and not relieved by a Bcl2 transgene. The DN2 arrest phenotype was extremely robust; we were able to document this developmental block even in T development assay cultures initiated with a single fetal liver derived precursor. The DN2-stage developmental block was confirmed using transduced Cre to induce deletion of a floxed Gata3 allele. Reduction of GATA-3 also increased PU.1 levels in pro-T cells, despite relatively normal T-lineage gene induction. To test roles of GATA-3 in exclusion of non-T fates, we forced expression of wild type, obligate repressor, and tamoxifen-inducible GATA-3 forms in precursors undergoing B and myeloid development. GATA-3 blocked B cell development via direct repression under conditions which remained permissive for myeloid development. Thus GATA-3 is required for at least two steps of T-cell progression to commitment, and restricts two different alternative fates by distinct mechanisms.
The role of MHC Ib in selecting functional CD8 T cell during acute LCMV infection (36.38)
Authors: He Xiao , Chen Lili , Jay David , Fairbanks Jared , Jensen Peter 
Published: January 2023

Abstract
Conventional MHC Ia & II molecules (H-2K/D/L & I-A/E in mouse) select α/β CD8 & CD4 T cell development, respectively. Although unconventional MHC class Ib, mainly encoded in H-2 Q, T & M regions in mouse, contains more than 20 MHC Ib genes, their functions remain largely unknown. We developed a mouse model, in which 3 genes, Kb, Db & CIITA were deleted, thus called triple knock-out (TKO) mouse. In TKO, CD8 T cells are selected only by MHC Ib. Previously we have shown that α/β CD8 T cells develop well in TKO. Functionally, we observe that TKO mice mount a protective immune response against in vivo acute infections of LCMV. The protection is CD8 T cell mediated & β2m dependent. Rapid production of IFNγ and Granzyme B is detected in expended CD8 cells. Intriguingly, TKO mice do not appear to mount a memory response to a second infection. We report that splenocytes, from in vivo LCMV primed TKO, can be restimulated in vitro, by LCMV infected peritoneal macrophages or bone marrow derived dendritic cells, to produce IFNγ, but little IL-2. This MHC Ib-dependent response appears to be partially mediated by Qa-2. Therefore we have established an in vitro system to analyze MHC Ib-restricted T cell responses to LCMV & we are attempting to map the MHC Ib restriction element(s) and target epitopes. Collectively, our results suggest that MHC Ib can select functional CD8 T cells & these T cells participate in, at least under some circumstances, protective immune responses to pathogens.