bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interleukin-4-mediated protection of primary B lymphocytes from apoptosis through Stat6-dependent up-regulation of glycolytic metabolism (94.16)
Authors: Gumina Maria , Bleiman Blair , Dufort Fay , Blair Derek , Chiles Thomas 
Published: January 2023

Abstract
Lymphocytes require extrinsic signals to maintain viability and this occurs, in part, through the up-regulation of anti-apoptotic proteins. Several recent studies have implicated cellular energy metabolism as an important component in preventing cell death following growth factor withdrawal. Interleukin 4 (IL-4) prevents death by neglect of naïve B lymphocytes; however, it is not known whether cellular glucose metabolism is crucial for survival by IL-4. We show herein that IL-4 triggers increased glucose uptake, glucose transporter (Glut1) expression, and glycolysis in splenic B lymphocytes. Inhibition of glycolysis results in apoptosis, despite the presence of IL-4. We demonstrate that insulin receptor substrate-2 and phosphatidylinositol 3-kinase activity are dispensable for IL-4-induced glucose utilization. By contrast, Stat6-deficient B cells exhibit impaired induction of Glut1 and glycolysis by IL-4. These findings establish that a key feature of IL-4 prevention of apoptosis in B cells is to promote glucose utilization via a Stat6-dependent pathway.
This work was supported by USPHS grant AI-49994.
Cholinergic agonists regulate monocyte chemoattractant protein-1 (MCP-1) production by activated endothelial cells: Involvement of the JAK/STAT3 pathway (94.29)
Authors: Chatterjee Prodyot , Sherry Barbara A , Metz Christine N 
Published: January 2023

Abstract
Numerous studies demonstrate the anti-inflammatory effects of cholinergic agonists, including nicotine. Investigations by our laboratory reveal that cholinergic stimulation inhibits endothelial cell activation, in part, through the NF-κB pathway. Monocyte chemoattractant protein-1 (MCP-1) is produced by endothelial cells and is a target of the JAK/STAT3 pathway. Human umbilical vein endothelial cells (HuVECs) were isolated and used to investigate the potential role of the JAK/STAT3 pathway in regulating MCP-1 production by activated endothelial cells following cholinergic stimulation. LPS induced MCP-1 expression in a dose-dependent manner. Soluble IL-6 receptor (IL-6sR), but not IL-6 itself induced significant MCP-1 expression. MCP-1 production following LPS or IL-6sR stimulation was mediated, in part, through the JAK/STAT3 pathway. Cholinergic agonists, nicotine and GTS-21, significantly reduced both LPS and IL-6sR-induced MCP-1 production. Cholinergic agonists also attenuated STAT3 phosphorylation following LPS and IL-6 stimulation. Finally, IL-6 blockade partially inhibits LPS-stimulated MCP-1 production and STAT3 phosphorylation. Together, our observations show that both LPS and IL-6sR induce MCP-1 production in endothelial cells, in part, through the JAK/STAT3 pathway that is inhibited by cholinergic mediators.
This work was supported by NIH NIGMS R01GM070727-02.
IL-2 inhibits its own production through a Blimp-1-dependent autoregulatory loop (94.4)
Authors: Gong Dapeng , Malek Thomas R. 
Published: January 2023

Abstract
IL-2 is essential for T cell expansion and effector cell development in vitro and for the development and maintenance of Treg cells in vivo. In our attempts to identify differentially expressed molecules dependent on IL-2 in activated T cells, DNA microarray analysis identified Blimp-1 as one such target. Blimp-1 is minimally expressed by naïve T cells, but is highly expressed by activated T cells. This expression mainly depends on IL-2, with lower Blimp-1 levels induced by IL-4 and IL-12. For activated T cells that expressed lower IL-2, Blimp-1 expression was higher, correlating Blimp-1 with the inability to properly produce lL-2. Activated T cells that were transduced with Blimp-1 produced lower IL-2 and inhibited an IL-2 reporter construct consisting of 8.4-kb upstream DNA sequence of the IL-2 gene, while granzyme B and CD25 expression were somewhat enhanced. These findings indicate that IL-2 inhibits its own production through a Blimp-1-dependent negative feedback loop while promoting a T effector cell phenotype. As Blimp-1-deficient T cells in vivo exhibit dysregulated T cell homeostasis and autoimmunity, our findings raise the possibility that improper regulation of IL-2 production may be a contributing factor to these pathological abnormalities.
Human Interferon Lambda-1 (IFN-λ1 / IL-29) modulates the Th1/Th2 response. (95.20)
Authors: Gallagher Grant , Jordan William , Eskdale Joyce , Srinivas Shekar , Pekarek Vera 
Published: January 2023

Abstract
Interferon lambda one ((IFN-λ1, IL-29) is a member of the Type-III interferon family, which contains three ligands: IFN-λ1, 2 and 3. These ligands use the same hererodimeric receptor, comprised of the CRF2-12 (IFN-λ1/IL-28Rα) chain and the CRF2-4 (IL-10Rβ) chains. Like their close relatives the Type-I interferons, IFN-λ1, 2 and 3 promote the phosphorylation of STATs 1&2, induce the ISRE3 complex, elevate OAS and MxA expression and exhibit anti-viral activity in vitro. Their use of the IL-10Rβ chain and their ability to phospohrylate STATs 3, 4 & 5 suggested that they might also exhibit immunoregulatory activity; their antiviral activity led us to hypothesize that this might be directed towards the Th1/Th2 system. Here, we demonstrate that IFN-λ1 altered the activity of human Th cells in three experimental systems: mitogen stimulation, MLR and the stimulation of naive T-cells by monocyte-derived dendritic cells(mDC). In general the inclusion of IFN-λ1 consistently led to a marked diminution of secreted IL-13 (15/16 donors) with occasional (5/16 donors) coincident, modest elevation of secreted IFN-γ. IL-13 secretion was 100-fold more sensitive to IFN-λ1 than was IFN-γ secretion. IFN-λ1 was able to alter Th-biasing and when naive T-cells were exposed to allogeneic mDC that had been developed in the presence of IFN-λ1, secreted IL-13 was again markedly and consistently reduced. These functions were independent of IL-10. Our data support a hitherto unsuspected role for IFN-λ1 in modulating the development of Th1 and Th2 cells, with an apparent emphasis on the diminution of IL-13 secretion in human in vitro cell culture.
Augmenting the biological activity of IL-15 with soluble IL-15Ralpha (95.21)
Authors: Rubinstein Mark P , Surh Charlie D , Sprent Jonathan , Goldrath Ananda W 
Published: January 2023

Abstract
IL-15 is thought to be normally presented in vivo as a cell-associated cytokine bound to IL-15Rα. We show here that the biological activity of soluble IL-15 is much improved after interaction with recombinant soluble IL-15Rα; after injection, soluble IL-15/IL-15Rα complexes rapidly induce strong and selective expansion of memory CD8+ cells and natural killer cells. Surprisingly, we also find that sIL-15/IL-15Rα complexes can drive conversion of naive T cells to memory T cells in vivo, without administration of antigen. This conversion is associated with an acquisition of effector function. The enhancing effect of IL-15Rα binding may explain why IL-15 normally functions as a cell-associated cytokine. Our results suggest the biological activity of IL-15 as a therapeutic reagent could be considerably enhanced by administration of preformed soluble IL-15/IL-15Rα complexes.
Interleukin-15 (IL-15) enhances, as well as increases the sensitivity of human CD8+ memory T cell subsets to stimulation by the pro-inflammatory cytokines IL-12 and IL-18 (95.3)
Authors: Smeltz Ronald B 
Published: January 2023

Abstract
Human memory CD8+ T cell subsets, termed central memory and effector memory, can be identified by expression of CD45RA, CD62L, and CCR7. Accordingly, functional differences have been described for each subset, reflecting unique roles in immunological memory. The common gamma chain cytokines IL-15 and IL-7 can induce proliferation and differentiation of human CD8+ T cell subsets, as well as increased effector functions. Here, we observed that addition of IL-15 or IL-7 to cultures of human CD8+ T cells profoundly enhanced the IL-12+IL-18 pathway of IFN-γ production. Importantly, IL-15 and IL-7 lowered the threshold concentrations of IL-12 and IL-18 required for induction of IFN-γ by 100-fold. Comparison of IL-15 and IL-7 demonstrated that IL-15 was superior for its ability to enhance IL-12+IL-18-induced IFN-γ, without evidence of synergy between IL-15 and IL-7. We also observed that IL-15 and IL-7-mediated enhancement of IL-12+IL-18-induced IFN-γ production was a functional property of effector-memory CD8+ T cells. Despite a lack of association between cell division and IFN-γ, loss of CD62L expression correlated well with acquisition of IL-12+IL-18-induced IFN-γ. Indeed, purified central memory T cells stimulated with IL-15 and IL-7 developed into effector memory T cells with potent IL-12+IL-18-induced IFN-γ. Thus, in addition to its known role in development of T cell memory, IL-15 may amplify CD8+ T cell effector functions by increasing sensitivity to pro-inflammatory cytokine stimulation.
This research was supported by a grant from the Jeffress Memorial Trust Fund and the Department of Microbiology at VCU.
IL-15 acts as a potent inducer of CD4+CD25high cells expressing forkhead box protein P3 (95.6)
Authors: Imamichi Hiromi , Sereti Irini , Lane H. Clifford 
Published: January 2023

Abstract
IL-15 is a member of the cytokine family that utilizes a common γ chain. While initially characterized as a T cell growth factor, predominantly involved in CD8+ T cell proliferation and survival, IL-15 is a pleiotropic cytokine that can affect both innate and adaptive immune responses. In the present study, we demonstrate that IL-15 increases expression of the alpha chain of the IL-2 receptor (CD25) and foxP3, a master control gene for the development and function of regulatory T cells, in human peripheral CD4+CD25− T cells without a need for additional antigenic stimulation signals. Side-by-side comparisons involving two other γc-cytokines: IL-2 and IL-7 revealed that the induction of CD25high and foxP3 expression was most prominent with IL-15 (median-fold increase: 46 and 53, respectively for CD25high and foxP3, p=0.0005) and IL-2 (32 and 42, p=0.0004). More modest effects were seen with IL-7 (18 and 12, p=0.0105). Despite levels of foxP3 expression comparable to that of conventional Tregs, these cytokine-induced CD4+CD25+foxP3+ cells exerted only weak suppressor activity. The current study has uncovered an unexpected function of IL-15 in the induction of CD4+CD25+foxP3+ cells from human peripheral T cells, thus helped to delineate and provided further insights into the pleiotropic nature of IL-15 beyond the regulation of CD8+ T cells.
Funded in part by NCI Contract N01-CO-12400.
TNF-α and oligodendrocyte apoptosis after cervical spinal cord injury (SCI) (96.11)
Authors: Schaal Sandra Marie , Marcillo Alexander Eduardo , Stagg Monica , Pearse Damien D. 
Published: January 2023

Abstract
The mechanism of SCI induced oligodendrocyte apoptosis is poorly understood. Tumor necrosis factor alpha (TNF-α) is a pleiotropic cytokine that is involved in inflammatory mediated cell death and thought to play a pivotal role in secondary pathology after SCI, including oligodendrocyte cell death. Utilizing a contusive SCI model in rat, we characterized the spatial, temporal, and cellular profiles of tnf-α expression and production acutely post-injury. Temporally, tnf-α mRNA, as assessed by RT-PCR, increased within 1 h and remained elevated through 24 h, albeit to a much lesser extent. In situ hybridization performed at 1 h revealed tnf-α mRNA throughout a cross-sectional area of the spinal cord. Though the tnf-α mRNA was diffusely distributed in the white matter, it was predominantly expressed in the gray matter. On the other hand, TNF-α protein increased by 1 h post injury and peaked at 4 h as measured by ELISA. At 1 h, 4 h, or 24 h, TNF-α immunoreactivity was present both within and adjacent to the injury epicenter; in the ventral gray matter and throughout the majority of the white matter. Using a cell specific marker (CC1) we found that oligodendrocytes were the principle TNF-α immunoreactive cell type after SCI as analyzed by stereological methods. These oligodendrocytes were also TUNEL positive, suggesting that TNF-α might be cytotoxic to oligodendrocytes acutely following cervical SCI.
CNS-Derived IL-4 is Essential for the Regulation of Autoimmunity and Induces an Alternative Activated Macrophage Phenotype in the CNS (96.20)
Authors: Ponomarev Eugene , Maresz Katarzyna , Dittel Bonnie N 
Published: January 2023

Abstract
Regulation of inflammation in the CNS is essential to prevent irreversible cellular damage that can occur in neurodegenerative diseases such as multiple sclerosis (MS). We investigated the role of IL-4 in regulating CNS inflammation using the animal model of MS, experimental autoimmune encephalomyelitis (EAE). We found that CNS-derived IL-4 was a critical regulator because mice with a deficiency in IL-4 production in the CNS, but not the periphery, had exacerbated EAE associated with a significant increase in the absolute number of infiltrating inflammatory cells. We also found that CNS-resident microglial cells in both the resting and activated state produced the protein Ym1/2, which is a marker of alternatively activated macrophages (aaMΦ), in an IL-4-dependent manner. This aaMΦ phenotype extended to the lack of nitric oxide (NO) production by activated microglial cells, which is a marker of classically activated macrophages (caMΦ). We also show that IL-4 induced the expression of Ym1/2 in peripheral macrophages, which also produce NO. Thus macrophages that migrate into the CNS exhibit a dual phenotype. These data indicate that IL-4 production in the CNS is essential for controlling autoimmune inflammation by inducing a microglial cell aaMΦ; phenotype. Macrophages that have undergone alternative activation have been shown to be important in tissue repair, thus our results suggest a new role for microglial cells in the regulation of inflammation in the CNS.
Differences in the expression of chemokine receptors by intrathecal B cells in B cell-dependent vs. B cell-independent EAE (96.25)
Authors: Gerber Kimberly , Lyons Jeri-Anne 
Published: January 2023

Abstract
B cells are implicated in the pathogenesis of multiple sclerosis. Data from the Experimental Autoimmune Encephalomyelitis (EAE) model are contradictory. We previously showed that myelin oligodendrocyte glycoprotein (rMOG)-induced EAE was B cell-dependent, while MOG35-55-induced disease was B cell-independent. Differences in the intrathecal B cell population may be relevant to the role of B cells in these two models. Higher numbers of B cells were detected in the CNS of rMOG-primed mice compared to MOG35-55-primed mice. Analysis of chemokine receptor expression by B cells isolated from the CNS of rMOG- and MOG35-55-primed mice was performed to further characterize the recruitment of B cells to the CNS during disease. Differences in the expression of CXCR1 and CCR4 was initially suggested by RT-PCR and confirmed by Q-PCR. Preliminary analysis of the cognate chemokines for these receptors indicated differences in the absolute expression as well as the expression profile of these ligands. Thus, differences in expression of factors important in B cell migration in rMOG- and MOG35-55-induced EAE may be relevant to the recruitment of pathogenic B cells in B cell-dependent EAE.
Latent Membrane Protein 1 of Epstein Barr Viruses Activates the PI3K Pathway to Induce Production of IL-10 (96.5)
Authors: Lambert Stacie L , Martinez Olivia M 
Published: January 2023

Abstract
Epstein Barr virus is a B lymphotrophic gammaherpes virus that is associated with multiple human malignancies. The EBV-encoded protein, latent membrane protein 1 (LMP1), is required for transformation of human B cells by EBV in vitro. LMP1 usurps multiple cellular signal transduction pathways to promote cell survival and to induce expression of cellular IL-10, an autocrine growth factor for EBV+ B cell lymphomas. However, the mechanisms by which LMP1 induces IL-10 are not understood. We previously showed that rapamycin, an mTOR inhibitor, could suppress IL-10 production by EBV-infected B cell lines. To test whether LMP1 activates rapamycin-sensitive signaling pathways to induce IL-10, we stably transfected EBV− BL41 cells to express a chimeric NGFR.LMP1 protein that mimics LMP1 signaling upon crosslinking. Treatment with either the p38 inhibitor SB203580 or with inhibitors that affect PI3K/AKT/mTOR signaling diminished (25–75% decrease) IL-10 production in EBV+ B cell lines. While induction of LMP1 signaling through NGFR.LMP1 resulted in phosphorylation of MAPK and PI3K pathway intermediates and production of IL-10, inhibition of either the p38 or PI3K/AKT/mTOR signaling axis markedly suppressed LMP1-mediated IL-10 production. These data indicate that p38 and PI3K/AKT/mTOR signaling contributes to LMP1-induced IL-10 production. Moreover, targeting these pathways may be an effective strategy to prevent IL-10 production and inhibit the growth of EBV+ B cell lymphomas.
Genetic variations in ITGA4 and susceptibility to MS (97.13)
Authors: O’Doherty Catherine , Ray Soma , Sun Chao , Carulli John , Vandenbroeck Koen , Antiguedad Alfredo 
Published: January 2023

Abstract
In recent years adhesion molecules have been studied in terms of their role in the pathophysiology and treatment of multiple sclerosis. Migration of leukocytes into tissues is facilitated by interactions between surface antigens such as Very Late Antigen (VLA4) (ITGA4/ITGA2 complex) and LFA1 and their endothelial ligands, VCAMs and ICAMs, respectively. Yednock et al (1) clearly pointed to a specific role for VLA4 in the pathogenesis of the animal model of MS, experimental autoimmune encephalomyelitis (EAE). VLA4 has been shown to be upregulated in patients and its expression has been related to both disease subtype and lesion development on MRI. Furthermore, Natalizumab, the monoclonal antibody to VLA4, has recently been licensed for use in MS. It reduced relapses and development of new lesions in clinical trials. The effective treatment of MS patients with Natalizimab points to the critical involvement of VLA4 in disease pathology and, lends further credence to the possibility that the benefits of IFNB may also be mediated to a large extent by suppression of VLA4 expression. We have examined 13 single nucleotide polymorphisms in ITGA4 (the gene encoding the á subunit of VLA4) in a cohort of 352 Basque MS patients and 235 healthy controls. Single marker and haplotype analysis were performed. Two SNPs, one in the promoter region and the other in intron 2, showed modest associations with susceptibility to MS.
This work was funded by Biogen Idec.
Microfluidic platform simulation of leucocyte adhesion to the endothelium (97.8)
Authors: Williams Vivienne , Kashanin Dmitry , O’Dowd Frank , Robinson Alexander John 
Published: January 2023

Abstract
Leukocyte adhesion to endothelial cell bound proteins, such as ICAM-1 and VCAM-1, is an initial step of the inflammatory response. We have developed an in vitro microfluidic system which mimics conditions found in blood vessels in vivo during an immune response. Using this system, we can record leukocyte adhesion levels under physiologically relevant flow conditions (e.g. 0–5 dynes/cm2).
The adhesion profiles of resting and PMA-stimulated peripheral blood lymphocytes (PBLs) were recorded, with respect to VCAM-1, ICAM-1, and BSA. Images at each shear stress level were captured using a digital camera, and analysed using our in-house Ducocell software package. Distinct morphological changes in PMA-stimulated PBLs, compared to non-stimulated cells, can be observed. These include a less rounded appearance of the PMA-stimulated PBLs, and evidence of “uropod” formation, which anchor the T cell to the endothelium as part of the migration process.
Levels of adhesion to VCAM-1 are high (80–90%, compared to control), but there appears to be little difference between the adhesion profiles of non-stimulated and PMA-stimulated PBLs. However, there is a distinct difference between the adhesion levels of non-stimulated and PMA-stimulated PBLs to ICAM-1, with PMA-stimulated cells showing a higher affinity for ICAM-1 than non-stimulated cells (approx. 70% and 40%, respectively). PBL adhesion to BSA is negligible.
A microfluidic platform providing improved throughput for neutrophil rolling and cell adhesion assays (97.9)
Authors: Ianescu-Zanetti Cristian , Blatz Andrew 
Published: January 2023

Abstract
A large number of processes underlying immune system biology are dependent on leukocyte adhesion and neutrophil rolling phenomena. In vitro, the control and quantification of the shear stress applied during such experiments is performed using laminar flow chambers. Despite their wide use, flow chambers suffer from low throughput and significant sterilization and setup times. Here we report on the development of a microfluidic platform that offers significant advantages over existing systems. Microfluidic designs enable the simultaneous assaying of a number of different shear regimes, while reducing reagent consumption. We present experimental data for cell adhesion and rolling neutrophil assays comparing traditional flow chamber experiments to microfluidic experiments at multiple shear force points, including time-lapse images. Biomimetic flow patterns and a wide range of shear protocols are also presented.
Lung eosinophils homing in homeostasis (98.2)
Authors: Sung Sun-sang Joseph , Lannigan Joanne A. , Rose C. Edward 
Published: January 2023

Abstract
Severe lung eosinophilia is a significant feature of the inflammatory response in asthma. Besides producing various secretory products that mediate tissue injury and leukocyte activation, eosinophils from inflamed tissues have been shown to present antigens to T cells. However, the phenotype and function of eosinophils in normal tissues is poorly characterized and their antigen presentation capacity controversial. With magnetic microbead selection against the major eosinophil markers, eosinophils from normal lung tissues have been enriched to 50% of the selected cells. Surface marker staining showed that eosinophils in homeostasis expressed low to undetectable levels of MHC class II and CD11c, two markers expressed by antigen presenting cells. However, they expressed the costimulation molecules CD80 and CD86. Unactivated eosinophils expressed integrins α4, α5, and α6 chains of the β1 family, high levels of the αL (CD11a) and αM (CD11b) chains of the β2 family, and αvβ3 and α4β7. They also expressed high levels of other adhesion molecules including SiglacF, ICAM-1, ICAM-2, PSGL-1, CD24, and CD44. Besides CCR3, eosinophils expressed CCR2 and CCR2-deficient mice showed marked reduction in lung eosinophils. These data indicate that in homeostasis, eosinophil antigen presentation requires the upregulation of MHC class II, whereas costimulation molecules are already on the surface. During antigen-mediated allergic airway inflammation, MHC class II expression on eosinophils is induced, rendering the eosinophils presentation-competent. Besides CCR3, eosinophil homing to lungs is mediated by CCR2 and a wide array of adhesion molecules.