bluuebunny/crossref_metadata_2025_split huggingface dataset:
Venetoclax (ABT-199), a potent and selective BCL-2 inhibitor, is efficacious in NZB/WF1 mouse model of lupus nephritis and reduces human lymphocyte lifespan in vitro (BA4P.127)
Authors: Grebe Kristie , Perper Stuart , O'Connor Liz , Schwartz Annette , Goess Christian , Hartman Dawna , Westmoreland Susan , Graff Candace , Souers Andrew , Leverson Joel , Elmore Steven , Olson Lisa , Wang Lichun 
Published: January 2023

Abstract
Proteins in the BCL-2 family are key regulators of apoptosis, or programmed cell death. We have conducted studies to evaluate a highly potent and orally available BCL-2 selective inhibitor, venetoclax (ABT-199), for efficacy and mechanism of action in the lupus prone NZB/W F1 mouse model and in human cells in vitro. Venetoclax treatment dose-dependently reduced the incidence of severe proteinuria and prolonged survival compared to vehicle controls and attenuated glomerulonephritis, tubular dilatation, immune cell infiltrates and IgG deposition in the kidney. Venetoclax mediated a significant reduction in the numbers of splenic T cells but conferred a preferential reduction in select B cell subsets. Interestingly, venetoclax did not impair the number of CD138+ long-lived plasma cells in the bone marrow, which was consistent with unaltered circulating anti-dsDNA titers in these animals. Venetoclax efficacy also correlated with a dose -dependent reduction of lymphocytes in peripheral blood of NZB/WF1 mice. Consistent with these findings, human lymphocytes from both healthy donors and systemic lupus erythematosus (SLE) patients treated with venetoclax in vitro have reduced lifespan. Taken together, these data underscore the essential role of BCL-2 in the pathogenesis of lupus and support further exploration of selective BCL-2 inhibition in autoimmune diseases such as SLE/Lupus Nephritis. The design, study conduct, and financial support for this research were provided by AbbVie.
Involvement of CD137L in peripheral nerve injury-induced neuropathic pain behaviors (CAM1P.161)
Authors: Malon Jennifer , Cao Ling 
Published: January 2023

Abstract
CD137L, also known as 4-1BBL, is a co-stimulatory molecule involved in many immune functions. CD137L-mediated signaling can promote monocyte/macrophage proliferation, migration, and production of proinflammatory factors; however, CD137L-mediated microglial responses have not been studied extensively. Here, we sought to investigate the role of microglial CD137L in the development of peripheral nerve injury-induced neuropathic pain using a murine model, spinal nerve L5 transection (L5Tx). First, compared to wild type (WT) controls, B6_CD137L knockout (KO) mice displayed significantly decreased L5Tx-induced mechanical and heat hypersensitivity starting at day 3 post-L5Tx until day 35 post-L5Tx when the experiment was terminated. To further determine the role of microglial CD137L, an anti-CD137L neutralizing antibody (Ab, clone TKS-1) was intrathecally injected into WT BALB/c mice daily from day 0 (before surgery) to day 7 post-L5Tx. The neutralizing Ab significantly reduced L5Tx-induced mechanical hypersensitivity but not the heat hypersensitivity. Our data indicate that spinal cord CD137L contributes to the maintenance of peripheral nerve injury-induced neuropathic pain-like behaviors. Future study will examine the involvement of microglial CD137L in L5Tx-induced lumbar spinal cord microglial and astrocyte activation.
Reciprocal regulation of c-Maf tyrosine phosphorylation by TEC and PTPN22 in TH2 cells (CCR3P.210)
Authors: Miaw Shi-Chuen , Liu Chih-Chun , Lai Chen-Yen , Lin Yu-Hsien , Chang Hui-Hsin , Ho I-Cheng 
Published: January 2023

Abstract
c-Maf plays an important role in regulating cytokine production in TH cells. It contains an N-terminal transactivation domain, which is connected to the C-terminal DNA binding domain through a hinge domain. c-Maf’s transactivation of IL-4 is optimized by phosphorylation at Tyr21, Tyr92, and Tyr131. However, the molecular mechanism regulating its tyrosine phosphorylation remains unknown. Here, we demonstrate that Tec kinase family member TEC, but not RLK or ITK, is a tyrosine kinase of c-Maf and that TEC enhances c-Maf-dependent IL-4 promoter activity. This effect of TEC is counteracted by PTPN22, which physically interacts with and facilitates tyrosine dephosphorylation of c-Maf thereby attenuating its transcriptional activity. We further show that phosphorylation of Tyr21/92/131 of c-Maf is also critical for its recruitment to the IL-21 promoter and optimal production of this cytokine by TH17 cells. Thus, manipulating tyrosine phosphorylation of c-Maf through its kinases and phosphatases can have significant impact on TH cell-mediated immune responses.
Dexamethasone suppresses IL-33-mediated mast cell activation (CCR3P.215)
Authors: Chernushevich Oksana , Paranjape Anuya , Ryan John 
Published: January 2023

Abstract
Dexamethasone, a glucocorticosteroid known for its anti-inflammatory and immunosuppressive activities, has been shown to inhibit IgE-mediated mast cell activation. The present research investigates the role of this drug in suppressing IL-33-mediated mast cell activation. IL-33 is a novel cytokine of the IL-1 family that elicits mast cells to produce inflammatory cytokines independently of IgE-FceRI signals. We have found that micromolar concentrations of dexamethasone are capable of suppressing IL-33-mediated cytokine production, regardless of genetic background. Dexamethasone also suppressed IL-33-induced cytokine production in mouse peritoneal mast cells. Inhibition was maximal when dexamethasone was added simultaneously with IL-33. Intracellular staining demonstrated that dexamethasone significantly reduces T1/ST2 expression in mast cells. However, cytokine suppression was shown to be independent of this downregulation of the IL-33 receptor. Thus, dexamethasone is capable of suppressing IL-33-mediated mast cell activation during the innate and adaptive immune responses. Further studies on the mechanism of dexamethasone are necessary for the understanding of those pathways and the development of more effective strategies for the treatment of inflammatory conditions.
RNA-binding protein HuR controls IL-2 homeostasis in activated T cells by regulating CD25 expression (CCR3P.202)
Authors: Glascock Jacqueline , Techasintana Patsharaporn , Magee Joseph , Atasoy Ulus 
Published: January 2023

Abstract
Posttranscriptional gene regulation by RNA binding proteins (RBPs) and their microRNA (miRNAs) partners is known to affect 50% of genes during T cell activation. The RBP, HuR regulates mRNAs by binding to AU-rich elements (AREs) in transcript 3’UTRs. Many Th2 cytokines and IL-2 possess AREs. We hypothesized that HuR may play a critical role in T cell activation and differentiation via regulation of IL-2 signaling pathways. To test this, we generated a conditional HuR KO mouse, distal-lck Cre ROSA HuRfl/fl, in order to ablate HuR prior to T cell activation. HuR KO T cells developed and egressed from thymus normally. However, they displayed an inability to shut off IL-2 and maintain stable cell surface expression of IL-2Ra (CD25). HuR KO T cells had striking increases in IL-2 mRNA and protein but decreases in Th2 cytokines. They also had proliferation defects, decreased p-stat5 and reductions in CD25 and prdm-1 transcription. We determined that HuR protein binds to IL-2 and CD25 mRNAs by two independent methods. CD25 mRNA stability was unchanged despite increased IL-2. We quantitated polysomal loading in HuR KO T cells compared to controls and discovered significant reductions in CD25 mRNA recruitment to heavy polysomes. Therefore, HuR regulates T cell activation and IL-2 homeostasis by controlling CD25 expression and enabling translational efficiency. We conclude that HuR plays an indispensible role in normal IL-2 homeostasis and T cell activation.
Mir-128-2 inhibits the development of B cell (HEM1P.218)
Authors: Yang Yi , Zhang Jinping 
Published: January 2023

Abstract
Accumulating evidence suggest that miRNAs play important roles in immune system. At present study, we first found that miRNA expression profiles were significantly different among vary immunocytes by microarray. Here, We focused on miR-128-2 which was highly expressed in DP T cells, but lower in other detected cells. In order to further study the function of miR-128-2 in lymphocytes development, we established the miR-128-2 chimeric and transgenic mice, confirmed by FACS analysis and PCR. We found B cell development appeared profoundly defects in both mice models, however, the development of T cells had no significant changes after over-expressing mir-128-2. Further analysis suggest that CLP was significantly higher in mir-128-2 TG mice compared to WT mice, but HSC and MMP cells were comparable in mir-128-2 TG mice and WT mice. Brdu and annexinV staining showed that over-expressing mir-128-2 did not affect the proliferation and apoptosis of CLP. These suggested that miR-128-2 inhibited the CLP cells to develop into B cell. Finally, we try to seek for the targets of miR-128-2 through software analysis and molecular biology methods. We found that miR-128-2 might down-regulate a group of genes including BCL11a, MALT1, A2b et. al. to inhibit B cell development confirmed by luciferase assay, real time PCR and western blot. Thus, our present study suggests that miR-128-2 mainly blocks CLP to differentiate into PreProB cells by down-regulating the BCL11a, MALT, A2b et al. genes.
Systemic adjuvant inflammation rapidly reorganizes bone marrow B cell populations, and their niche, to shape B cell function and repertoire development (HEM1P.224)
Authors: Moreau Joshua , Paige Christopher 
Published: January 2023

Abstract
Maturing B cells migrate through the organized bone marrow parenchyma toward sinusoid vessels before entering circulation. Niche components rigorously control this process. Infecting mice with pathogens, or injecting inflammatory agents, is known to produce perturbations in B lymphopoiesis, however; it is poorly understood how the B cell niche responds to these challenges. Moreover, the impact on subsequent B cell function remains unknown. Here, we report that injection of Freund’s incomplete adjuvant in mice induced a large and rapid (within hours) mobilization of B cells to the bone marrow, mediated by increased sensitivity to the chemokine CXCL12. Notably, localization within the marrow is altered with all B cell subsets showing a reduced affinity for the sinusoids. Reorganization of B cell positioning occurred together with relevant modification of their supportive niche including increased expression of BAFF and MIF mRNA, expansion of DX5+Thy1loAPRIL+ basophils, and elevation of several important cytokines. This altered marrow milieu may have a role in shaping B cell fate, as we observed a decrease in lambda light chain usage in immature cells. Further, disruption of homing dynamics with a CXCR4 inhibitor attenuated the formation of IgM secreting cells. Together these results demonstrate an exquisite sensitivity of the marrow B cell compartment to peripheral inflammation and suggest that initial inflammatory signaling regulates subsequent B cell effector responses.
Pre-B cells suppress RAG expression in response to DNA double-strand breaks (HEM1P.225)
Authors: Fisher Megan , Bassing Craig 
Published: January 2023

Abstract
The ability of the Rag1/Rag2 (RAG) endonuclease to assemble antigen receptor (AgR) genes is essential for adaptive immunity. However, aberrant induction or repair of RAG-induced DNA double strand breaks (DSBs) can lead to oncogenic AgR translocations. We have previously shown that RAG-induced DSBs in pre-B cells activate the ATM kinase to prevent RAG cleavage of the homologous allele, and to inhibit expression of Rag1 and Rag2. These breaks also suppress expression of Gadd45α, which promotes Rag1 and Rag2 transcription. Since DSBs induced by ionizing radiation (IR) signal through ATM to increase Gadd45α expression in all other cell types including mature B cells, these results indicate that RAG DSBs and/or pre-B cells use unique mechanisms to regulate Gadd45α expression. We now demonstrate that IR-induced DSBs in pre-B cells signal through the ATM kinase to down-regulate Gadd45α expression. We also demonstrate that these IR DSBs signal ATM-dependent suppression of Rag1 and Rag2 transcription, without affecting stability of Rag1 or Rag2 mRNA. Because the presence of multiple DSBs in a cell greatly increases the risk of translocation, we hypothesize that pre-B cells have developed a unique developmental stage-specific DSB response to suppress the induction of RAG DSBs in the presence of other DSBs. Our ongoing studies will determine the role of this pre-B cell specific DSB response in suppressing AgR translocations and enforcing mono-allelic assembly of immunoglobulin genes.
HEB plays a critical role in the installment of IL-17 program in fetal thymic γδ T cells (HEM2P.231)
Authors: In Tracy , Anderson Michele 
Published: January 2023

Abstract
IL-17 producing γδ T cells that arise during fetal development form an integral part of the immune system of various lymphoid and mucosal tissues in mice. Here we aim to better understand the transcriptional networks that drive the functional programming of IL-17 producing γδ T cells. Specifically, we investigated the role of the transcription factor HEB in this process using mice that carry the HEB null allele. To evaluate the role of HEB in fetal γδ T cell development, we utilized fetal thymic organ culture from HEB+/+ and HEB-/- embryos at E14. We found that the absence of HEB did not have a significant impact on γδ T cell numbers, and that all functional subsets of γδ T cells were present as indicated by expression of CCR6, CD27, CD44 and CD62L. However, the fetal thymic γδ T cells from HEB knockout mice exhibited a profound deficiency in their ability to produce IL-17, especially in the CD44+ γδ T cell population. In addition, HEB-deficient fetal γδ T cells expressed significantly lower levels of SOX13 and RORγt, factors associated with IL-17 producing γδ T cells, while expression of IFNγ, Tbet and Egr3 were unchanged. Furthermore, HEB conditional knockout mice on a Vav-Cre background had significantly lower frequencies of IL-17 producing γδ T cells in the spleen, lymph nodes and lungs. Collectively, our work shows for the first time that HEB plays an important role in installing the IL-17 program in γδ T cells.
Bim expression in early DP thymocytes limits development of TCRαβ+DN thymocytes and CD8αα intestinal intraepithelial lymphocytes (HEM2P.243)
Authors: Li Kun-Po , Roy Eron , Cuda Carla , Grimes H , Perlman Harris , Hildeman David 
Published: January 2023

Abstract
Thymocyte negative selection is thought to cull T cells with strong self-reactivity and maintain T cells with non-self-specificity. While initial work implicated Bim as a critical mediator of thymocyte negative selection, its role in the process has been controversial. Thus, we re-examined thymocyte development in mice with global and T cell specific loss of Bim. We found that global Bim-/- mice had a striking accumulation of DN4 thymocytes expressing high levels of surface TCR, which failed to generate DP cells in OP9-DL1 culture. Interestingly, CD4Cre-driven deletion of Bim promoted DN4 cells, but later, dLckCre-driven deletion of Bim did not. Combined, these data show that the aberrant DN4 cells in Bim-/- mice were once DP cells, but had lost co-receptor expression. Investigating the fate of these cells, we found that splenic DN T and gut CD8αα T cells were increased in Bim-/- mice. While IL-15 is critical for both DN/CD8αα T cells, the additional loss of Bim partially restored DN/CD8αα T cells in IL-15 deficient mice, suggesting that IL-15 signals antagonize Bim to promote DN T and CD8αα T cells. Finally, TCR or IL-15 signals were sufficient to promote expression of both CD8αα and the gut homing receptor CCR9 on splenic DN T cells. In conclusion, these data further delineate the role of Bim on thymocyte fate and DN/CD8αα T cell development; and show that dLckCre-Bimf/f mice can isolate the effects of Bim on peripheral T cell survival without affecting thymic selection.
Developmental changes in Tcrb architecture are independent of thymocyte proliferation (HEM2P.248)
Authors: Majumder Kinjal , Koues Olivia , Rupp Levi , Kyle Katherine , Collins Patrick , Bassing Craig , Oltz Eugene 
Published: January 2023

Abstract
Assembly of the genetic components of the adaptive immune system by V(D)J recombination must be tightly controlled to prevent chromosomal translocations that cause genomic instability. This control is exerted at the level of tissue, stage and allele specificity by genetic and epigenetic mechanisms. Changes in chromosome conformation is one of the mechanisms of epigenetic control. In DN thymocytes, the Tcrb locus adopts a conformation which brings together distally located V, D and J gene segments by looping out intervening DNA to facilitate rearrangement. As the thymocyte proliferates and proceeds to the DP stage of development, the distal ends of Tcrb dissociate from one another. We applied high-throughput 4C-sequencing strategy to study the developmental changes in germline Tcrb looping at greater resolution. Characterization of the interactomes of multiple viewpoints revealed that: (i) the D-J region interacts with central Vβs in DP thymocytes despite losing distal Vβ contacts and (ii) the distal Vβs maintain a unique architecture in DP thymocytes. This conformation of Tcrb is maintained in DP thymocytes independent of proliferation, suggesting that lineage-specific factors can stabilize chromosome loops. Current studies are focused on deciphering the molecular mechanisms that regulate these developmental changes in Tcrb looping including structural protein binding and perturbation of chromatin barrier elements.
Cardiac myosin and the Th17 immunophenotype in human autoimmune myocarditis (HUM1P.252)
Authors: Cunningham Madeleine , Myers Jennifer , Cooper Leslie , Kem David , Stavrakis Stavros , Shevach Ethan , Fairweather DeLisa , Stoner Julie , Cox Carol 
Published: January 2023

Abstract
The immunopathogenesis of autoimmune myocarditis and dilated cardiomyopathy (DCM) in humans is unpredictable and pathogenic mechanisms undefined. The aim was to investigate potential autoimmune mechanisms in human autoimmune myocarditis and heart failure. Our evidence for a Th17 immunophenotype in human myocarditis/DCM(n=40) included reduced CD4+CD25+FoxP3+ Tregulatory cells(0.0006), elevated Th17 cells(0.0008) and increased Th17-promoting cytokines(IL-6 and TGF-beta). Th1(IFNγ+) cells were not significantly elevated(p=0.7184). IL-6(&lt;0.0001), TGF-beta(0.0001), IL-23(0.0003), and GM-CSF+ T cells(0.0006) were significantly elevated. IL-17A+ cells were prevalent in biopsies from human myocarditis, non-recovery associated with persistence of IL-17A cytokines and heart failure associated with elevated Th17 cells in peripheral blood. To identify a potential mechanism to promote a Th17 pro-inflammatory pathogenesis in the heart, novel TLR ligands of human cardiac myosin (HCM) (peptides S2-16 and S2-28) promoted myocarditic PBMCs/CD14+ monocytes to produce Th17 cytokines(IL-23 p=0.0289; IL-6,TGF-beta p&lt;0.0001). TLR ligand stimulation of PBMCs/ monocytes was abrogated by antibodies blocking TLR2, IL-6, IL-17A/23, and GM-CSF. Our translational study suggested human myocarditis/DCM may be controlled by a Th17 phenotype including reduced Treg, elevated Th17 cells, and pro-inflammatory and pro-fibrotic Th17 cytokines triggered by cardiac myosin TLR ligands potentially released from heart.
PARP inhibitor, olaparib ameliorates acute lung injury by modulating oxidative stress and NF-κB mediated inflammatory response in mice (HUM1P.265)
Authors: Naura Amarjit , Kapoor Kunal , Singla Esha , Sahu Bijayani 
Published: January 2023

Abstract
We have previously shown that poly (ADP-ribose) polymerase (PARP)-1 gene deletion in mice provide robust protection against lung inflammation in the context of asthma and acute lung injury. Olaparib is a potent new generation PARP inhibitor that has been approved for human testing in cancer patients. The present work was designed to evaluate its beneficial potential against LPS-induced acute lung injury upon intra-tracheal administration of the endotoxin in mice. Administration of olaparib at different doses, 30 min after LPS treatment showed that single injection of the drug effectively reduced the total number of inflammatory cells particularly neutrophils in the lungs. This was associated with reduced pulmonary edema as the total protein content in bronchoalveolar fluid was found to be decreased substantially. The drug restored the LPS mediated oxidative stress toward normal in lungs as assessed by measuring malondialdehyde and GSH levels. Finally, RT-PCR data revealed that olaparib downregulates the LPS induced expression of NF-κB dependent genes namely TNF-α, IL-1β and VCAM-1 in the lungs without altering the expression of total p65NF-κB. Overall, the data suggests that olaparib possesses a strong therapeutic potential against LPS induced lung injury in mice. Given the fact that olaparib is approved by FDA for human testing, our findings can pave the way for testing of the drug on humans inflicted with acute lung injury.
Th1, Th2, Th17 and Treg levels in umbilical cord blood of preeclamptic women. (HUM1P.300)
Authors: Soto-Vega Elena , Solleiro-Villavicencio Helena , Vargas-Rojas Maria 
Published: January 2023

Abstract
Preeclampsia is one of the major causes of maternal and neonatal mortality. The immune system during pregnancy has to maintain the tolerance to the fetus, thus changes in the cytokine balance may result in a disturbed pregnancy. T helper cells play an important role in the modulation of the immune system. Many studies have been performed to study the T cell composition in different compartments during pregnancy, although this is the first study in which the levels of Th1, Th2, Th17 and Treg cells are evaluated in umbilical cord blood from normal and preeclamptic women. We did not find any difference in Th1 and Th17 levels between groups, but Th2 and Treg levels were significantly diminished in preeclamptic women. Even if we found no differences between pro-inflammatory profiles, their counterparts Th2 and Treg are diminished, suggesting that the balance is broken encouraging the development of an exacerbated inflammatory response. Our results show that there is a shift, not only in the Th1/Th2, but also in the Th17/Treg balance, favoring skewness towards a proinflammatory status in preeclampsia, so the innate and adaptative immune system are involved in the development of an exaggerated maternal systemic inflammation.