bluuebunny/crossref_metadata_2025_split huggingface dataset:
PD20, a novel regulator of lymphocyte responses and immune homeostasis. (130.5)
Authors: Sun Honghong , Gong Shunyou , Hilliard Anja , Chen Youhai H. 
Published: January 2023

Abstract
Experimental autoimmune encephalomyelitis (EAE) is an animal model for human multiple sclerosis (MS). Using a high-throughput gene microarray technology, we identified a novel gene, designated programmed cell death 20 kD (PD20) from inflamed spinal cord of mice with EAE. PD20 encodes a polypeptide of 184 amino acids with an estimated molecular weight of 20 kD. It is preferentially expressed in lymphoid tissues and inflamed but not normal nervous tissue. Germ line mutation of the PD20 gene induced a systemic inflammatory disease characterized by multi-organ inflammation, splenomegaly, heightened inflammatory cytokine production and eventual death of the animals. The disease can be significantly accelerated by a single injection of endotoxin. In vitro, PD20-deficient T cells are hypersensitive to TCR stimulation but are resistant to activation-induced cell death. Importantly, PD20 is able to interact with caspase-8 and is capable of enhancing Fas ligand-induced cell death. These results establish that PD20 is a novel caspase-8-interacting protein that regulates apoptosis, lymphocyte responses and immune homeostasis.
Inhibitory oligonucleotide (IN-ODN) structure-function relationships: species and backbone differences (131.19)
Authors: Ashman Robert F. , Goeken Adam , Lenert Petar 
Published: January 2023

Abstract
The recognition of CpG motifs in bacterial DNA by Toll-like Receptor (TLR9) is a key function of the innate immune system, announcing the presence of an infection. We have previously developed synthetic oligonucleotide (ODN) inhibitory of TLR9 function with phosphorothioate (S) backbones conforming to the sequence n CC n not C not C nn GGG nnnn where n is any base. Here we show that these sequence requirements generate IN-ODN when the backbone is phosphodiester (O) also. In general, whereas stimulatory O-ODN are 2 logs less potent than their S-backbone congeners IN-ODN are 3 logs less potent. Changes in the critical bases diminish activity with either backbone; however, when the backbone is S, such changes diminish activity of stimulators much less, and inhibitors much more, than if the backbone is O. Certain end truncations enhance activity and even the minimal sequence CCTGGAGGG retains substantial inhibitory activity. Human-mouse relative activity comparisons tend to be similar between critical base variants, but may differ substantially between end truncation variants. HEK cells transfected with mouse or human TLR9 and an NFkB binding luciferase promoter respond to much lower concentrations of IN-ODN than do primary cells of either species. The neutral sequences with S backbones developed for mouse cells are also neutral with O backbones and with human cells.
Supported by AI04737405 from NIH
MDX-1100, a fully human anti-CXCL10 (IP-10) antibody, is a high affinity, neutralizing antibody that has entered Phase I clinical trials for the treatment of Ulcerative Colitis (UC). (131.20)
Authors: Kuhne Michelle , Preston Benjamin , Wallace Steven , Chen Sharline , Vasudevan Geetha , Witte Alison , Cardarelli Pina 
Published: January 2023

Abstract
The chemokine, CXCL10 or interferon γ inducible protein-10 (IP-10) is a chemotactic cytokine for activated T cells and monocytes and plays an important role in migration of cells into sites of inflammation. The receptor for CXCL10, CXCR3, is expressed by activated T cells, eosinophils, NK, and endothelial cells. CXCL10 levels are elevated in ulcerative colitis (UC) amongst other inflammatory diseases. In preclinical animal models of UC, antibodies against CXCL10 have been shown to modify disease progression. Medarex, Inc. has developed a fully human monoclonal antibody (MDX-1100) that binds selectively to CXCL10. This antibody binds to the ligand with high affinity and effectively competes for ligand binding to CXCR3 expressing cells. MDX-1100 blocks CXCL10 induced calcium flux and cell migration with an estimated IC50 in the low nM range. In order to identify potential pharmacodynamic markers for CXCL10 activity, gene chip analysis was performed with RNA purified from CXCL10-stimulated human PBMCs. We identified cell surface receptors, intracellular and soluble markers that are CXCL10 responsive and confirmed the induction of a subset of these genes using quantitative RT-PCR. In summary, MDX-1100, an antibody that binds and neutralizes the activity of CXCL10 is predicted to reduce disease severity in patients with UC and thus a Phase I clinical trial has been initiated.
Histone acetyltransferase p300 acts as a potent transcriptional co-activator for Pax5 (35.29)
Authors: He Ti , Yu Zhihong , Kwon Hyoung , Toleman Clifford A , Kudlow Jeffrey , Zhang Zhixin 
Published: January 2023

Abstract
Pax5/BSAP is an essential regulator for B lineage cell development and function. Pax5 activates the transcription of many B lineage specific genes, such as Cd19, Mb-1 and Blnk; controls the B lineage specific IgH VH→DJH recombination and meantime, represses the expression of other lineage inappropriate genes, such as Notch1, M-csfr, G-csfr, and Ccl3, to restrict B lineage cell development. It becomes important to understand how the Pax5 activity is regulated to exert these different functions. Here, we show that co-expression of histone acetyltransferase p300 dramatically enhances Pax5-mediated activation of a Pax5 responsive luciferase reporter construct. The p300-mediated enhancement is dependent on the intrinsic histone acetyltransferase activity of p300 and the activation domain of Pax5. Pax5 interacts with p300 in vitro and forms a complex with p300 in B lineage cells. Recombinant full length p300 purified from baculovirus or 293 cells directly acetylates purified Pax5 in vitro and overexpression of p300 induces Pax5 acetylation in 293 cells. Mass spectroscopy-based analysis identified that multiple lysine residues within the Pax5 DNA binding domain were acetylated upon coexpression of p300. Mutations of lysine 67, 87, 89, 103, and 142 residues on Pax5 attenuate p300-mediated enhancement. These results suggest that p300 acts as a potent co-activator for Pax5-mediated transcription function through acetylation of the Pax5 DNA binding domain.
Epigenetic Control of Killer Immunoglobulin-like Receptor (KIR) Expression in T cells (35.40)
Authors: Li Guangjin , Weyand Cornelia M. , Goronzy Jorg J. 
Published: January 2023

Abstract
Killer immunoglobulin-like receptors (KIR), usually restricted to NK cells, are expressed on T-cells with increasing age and contribute to age-related diseases. We examined the epigenetic changes associated with KIR expression. In T cells with absent KIR2DL4 transcripts such as the Jurkat cell line or CD4+CD28+ T cells, CpG sites of the proximal KIR2DL4 promoter were hypermethylated. DNMT inhibition in such cells induced low but appreciable KIR2DL4 transcription. DNMT inhibition was strikingly more effective in T cells that had some constitutive KIR2DL4 transcription, such as CD4 and CD8 T cells from elderly adults or the CD4+CD28−T cell line HUT78. Such cells had evidence of partial promoter demethylation; however, although the proximal nucleosome was partially relaxed, the only histone modification was an increase in dimethylated H3-Lys 4. In contrast, NK cells had a fully demethylated KIR2DL4 promoter and the full spectrum of histone modifications indicative of active transcription with H3 and H4 acetylation, di- and trimethylated H3-Lys 4, and reduced dimethylated H3-Lys 9. These results suggest that early steps of promoter activation in T cells include selective increase in H3-Lys 4 dimethylation and limited DNA demethylation. This partially accessible promoter is sensitive to DNMT inhibition, which is sufficient to induce full transcription without further histone acetylation and methylation.
Supported by NIH grant AG 15043
Phosphorylated mRNA binding proteins associated with NE/cAMP/PKA-mediated mRNA decay in S49 T lymphocytes (35.43)
Authors: LaJevic Melissa Dawn , Koduvayur Sujatha , Graf Lloyd H , Cohen Rhonna L , Chambers Donald A 
Published: January 2023

Abstract
We have shown that norepinephrine (NE) mediates the β-adrenergic/cAMP/PKA-dependent mRNA destabilization of mRNAs in T lymphocytes. Our goal is to elucidate the mechanism(s) involved in NE mediated Thy-1 mRNA decay in S49 T lymphoma cells. NE signal transduction in S49 cells activates PKA which could lead to the phosphorylation of effector proteins involved in ARE mediated message decay. Previous results have identified AUF1, HuR, KSRP, TIAR, and Hsc70 as candidate effector proteins. This study analyzes phosphoproteins from S49 cells and the effect of NE treatment (150μM, 30 min) on phosphorylation. NE induced phosphorylation of PKA substrate in S49 cytoplasmic extract was identified at 47, 55, 60, and 105 kD via immunoblotting. Of the candidate proteins that correlate in size, only AUF1 and Hsc70 were detected in phosphoprotein isolated from S49 whole cell extract via affinity chromatography; however, their level of phosphorylation appears unchanged with NE treatment. To determine if other binding proteins may be PKA targets, a pull down assay using a biotinylated 170 nucleotide ARE rich Thy-1 probe and 32P metabolically labeled cytoplasmic protein (100μCi/ml, 30 min) isolated at least five phospho-binding protein, two of which, 60 kD and 105 kD, increased with NE treatment. These results characterize phosphoproteins that function in the NE-mediated decay of Thy-1 and other mRNAs.
Supported by NIH
Replicating dendritic cells transmit internalized antigen to progeny for presentation (36.19)
Authors: Diao Jun , Winter Erin , Chen Wenhao , Cattral Mark S 
Published: January 2023

Abstract
During steady state conditions spleen dendritic cells (DC) turnover every 2–3 days. Recent evidence indicates that local proliferation of spleen DC contributes substantially to their homeostasis. We and others have shown that the majority of these dividing DC arise from a distinct population of replication-competent immediate conventional DC precursors (cDCp). cDCp generate exclusive conventional DC that continue to replicate for several generations. The objective of this study was to determine whether DC can replicate after capturing antigen, and whether replication provides a mechanism for disseminating antigen to their progeny. Real-time confocal microscopy revealed that replicating DC internalize and directly transfer model antigens to successive generations of progeny. Soluble protein antigen (ovalbumin) inherited by DC progeny in vitro or in vivo was presented effectively to OT-1 T cells. Other mechanisms of inter-DC antigen transfer (e.g. exosomes, cell membrane exchange) did not contribute to antigen acquisition by DC progeny. Our results suggest that replication of antigen-bearing DC is a fundamental behavior and may provide a novel mechanism for increasing the frequency of antigen-bearing DC, and duration of antigen presentation.
New STAT6 inhibitors with potential therapeutic application in asthma (37.1)
Authors: Zamorano Jose , Cortes Jose R , Perez-G Moises , Fernandez Carlos , Gonzalez-Nunez Maria A 
Published: January 2023

Abstract
Asthma is an inflammatory disease that represents an important health problem. The origin of this disease seems to be due to an immunological anomalous response to apparently innocuous antigens. In the last years, an important number of molecular intermediaries have been found to be implicated in the progression of this disease. The importance of these mediators is determined by the fact that they are actively being investigated as potential therapeutic targets. In this regard, numerous evidences indicate that Interleuquina-4 (IL-4) and IL-13 have a central role in the development of asthma through the activation of the factor of transcription STAT6. In a previous study, our group demonstrated that antiinflammatory doses of salicylates inhibited the activation of this factor of transcription. The objective of the present study was to obtain new inhibitors of STAT6 derived from salicylates by performing sequential chemical modifications. With this approach, we obtained several compounds with a planar and rigid structure, and a specific pattern of hydroxyl groups that were able to inhibit the activation of STAT6 with great efficiency. Among the obtained compounds, the named JRC688 and JRC706 inhibited the activation of STAT6 at concentrations of 20 μM. To investigate the effect of these compounds in asthma, we chose JRC706 since it had little effect on cell viability, suggesting it could have less undesirable effects. The found results indicate that the administration of JRC706 could prevent the inflammatory response in asthma. Thus, infiltrated inflammatory cells in lung and bronchoalveolar fluids, mucus production, hyperplasia of globet cells, and levels of IgE were greatly diminished in mice treated with JRC706 with regard to untreated asthmatic controls. These findings indicate that JRC706 is a good compound to investigate in asthma.
Blocking OX40L Function Inhibits TSLP-Induced Atopic Disease In Lung And Skin (37.11)
Authors: Seshasayee Dhaya , Zhou Meijuan , Shu Jean , Suto Eric , Zhang Juan , Lee Wyne Pun , Seeber Stefan , Fuentes Maria , Hymowitz Sarah , Martin Flavius 
Published: January 2023

Abstract
OX40L-OX40 interactions have been shown to be important for development of Th2-mediated diseases such as asthma through the use of mouse models in vivo. Thymic Stromal LymphoPoietin (TSLP), an IL7-like cytokine, has been shown to potently induce atopic immune responses and is highly expressed at sites of allergic inflammation. While the ability of TSLP to effectively induce an atopic immune cascade has been demonstrated through the use of transgenic mice, direct downstream in vivo mediators have not been identified. In our current study, we show that OX40L is a critical mediator of TSLP-mediated Th2 responses not only in vitro but also in vivo. TSLP strongly induces OX40L expression on dendritic cells and blocking a-OX40L antibodies efficiently inhibit Th2 responses induced by TSLP in vitro, and in the lung and skin in vivo. Inhibition of OX40L function was also very efficacious in decreasing Th2 cytokines and antigen-specific IgE in a mouse model of OVA-induced asthma. The use of blocking a-OX40L antibodies thus presents an effective strategy for the treatment of allergic disease associated with dysregulated Th2 immune responses.
The source of research support is Genentech, Inc.
Global patterns of gene expression following inhalation of toluene diisocyanate suggest development of allergic rhinitis (37.4)
Authors: Johnson Victor J , Kashon Michael L , Yucesoy Berran , Luster Michael I 
Published: January 2023

Abstract
Rhinitis is often a comorbid and preceding disease in workers afflicted with toluene diisocyanate (TDI) asthma. We hypothesized that TDI rhinitis is driven by an allergic immune response rather than innate inflammation. Global patterns of gene expression were examined via microarray in a murine model of TDI rhinitis and compared to acute nasal exposure to lipopolysaccharide (LPS). Histological analysis showed extensive inflammation of the nasal mucosa with eosinophils and neutrophils dominating in the TDI and LPS models, respectively. Effector cell recruitment correlated with expression of chemokines directed towards eosinophils in TDI rhinitis and neutrophils in LPS rhinitis. Genes involved in antigen processing/presentation, lymphocyte activation/regulation, cytokine-receptor signaling and humoral immunity were differentially regulated in TDI rhinitis. In contrast, inflammatory, innate immune and defense responses as well as response to pathogens/biotics were over represented in LPS rhinitis. These results indicate that exposure to TDI results in an antigen-driven immune response in the nasal mucosa suggestive of allergic rhinitis.
This work was supported in part by an NIEHS IAG (Y1-ES-0001 - Immunotoxicity of Workplace Xenobiotics)
The findings and conclusions have not been formally disseminated by NIOSH and should not be construed to represent any agency determination or policy.
Novel therapy for allergic disease through gene silencing of CD40 (37.7)
Authors: Suzuki Motohiko , Zhang Xusheng , Zheng Xiufen , Li Mu , Vladau Costin , Chen Dong , Garcia Bertha , Min Wei-Ping 
Published: January 2023

Abstract
Gene silencing is a potent, selective, and easily-inducible method for specifically blocking expression of desired genes. Gene silencing strategies have been successfully tested in animal models of various diseases. However, the therapeutic potential of gene silencing allergic diseases has not yet been reported. CD40 is an important costimulatory molecule that plays a critical role in immune responses. We attempted to develop a new therapy for allergic diseases through gene silencing using a short hairpin siRNA-expressing vector (shRNA) specific to CD40. The allergic mouse model was made by intraperitoneal immunization with ovalbumin (OVA), followed by intranasal challenges with the same antigen. CD40 shRNA was administered before or after OVA immunization. CD40 shRNA treatment reduced CD40 expression in splenic DC, remarkably reduced nasal allergic symptoms, and decreased nasal eosinophilia. The OVA-specific T cell response was inhibited after CD40 shRNA treatment. Additionally, anti-OVA specific IgE was significantly decreased in CD40-shRNA treated mice, as detected by ELISA. The production of IL-4 and IL-5 was suppressed in the CD40-shRNA treated mice. Finally, CD40 shRNA facilitated the generation of regulatory T cells, in particular, the CD4+CD25+Foxp3+ subset of cells. This study, for the first time, has demonstrated a novel therapy for allergic disease through gene silencing.
Effects of P-glycoprotein Deficiency on Epithelial Cell Responses to Toll-like Receptor Ligands (40.2)
Authors: Staley Elizabeth M , Dimmitt Reed , Lorenz Robin G 
Published: January 2023

Abstract
P-glycoprotein (P-gp), the product of the multi-drug resistance gene (MDR), is an ATP-dependent transmembrane pump. In intestinal epithelial cells, loss of P-gp affects tight junction proteins, and increases membrane permeability. Deficiency of this protein results in colitis in conventionally housed mice. It is believed P-gp functions to extrude xenobiotic stimuli from the intracellular environment, preventing contact with intracellular pathogen receptors. Therefore, we hypothesized that P-gp deficiency may cause colitis due to stimulation of intracellular toll-like receptors (TLRs) by excessive accumulation of bacterial antigens. HEK cells stably transfected with TLR expression plasmids were electroporated and treated with siRNA targeted for the MDR1a gene. Cells were then stimulated with TLR ligands. Inflammation was measured as IL8 secretion, an inflammatory cytokine produced by epithelial cells, and quantified by cytokine ELISA.
P-gp deficient HEK cells expressing TLR9 produced 4.9 +/− 1.3 ng/ml IL8 when treated with stimulatory CpG-OGNs, cells wild type for P-gp expression only produced 1.6 +/−.44 ng/ml IL8. No significant difference in IL8 production was detected in cells expressing TLRs 4 or 5 when treated with LPS or flagellin respectively.
P-gp may function in intestinal epithelial cells to extrude stimulatory bacterial CpG and reduce unwarranted inflammatory response.
Supported by NIH grants T32AI07051; NIHP01DK071176; & R24 DK064400
The mast cell activator Compound 48/80 is a potent adjuvant for intradermally-administered anthrax protective antigen. (41.12)
Authors: McGowen Afton , Paraggio Nicole A. , Sobel Ashley E. , Kirwan Shaun M. , Shelburne Chris P. , Pizzo Salvatore V. , Abraham Soman N. , Staats Herman F 
Published: January 2023

Abstract
Recent studies have reported that mast cells play a role in the trafficking of dendritic cells and lymphocytes to lymph nodes draining sites of infection. To test the hypothesis that mast cell activators could be used as vaccine adjuvants, C3H/HeN mice were intradermally vaccinated in the ear on days 0 & 21 with 0.5 μg recombinant anthrax protective antigen (rPA) alone or combined with 30 μg compound 48/80 (C 48/80), a mast cell activator. CpG oligodeoxynucleotide (CpG ODN, 1 μg) was used as a control. Ear swelling was monitored 24 hours after immunization to measure injection site inflammation. Mice immunized with rPA alone had an average ear thickness (mm) of 0.0059 while mice immunized with rPA + C 48/80 had an ear thickness of 0.0426 (p &lt; 0.001 vs rPA alone). Mice immunized with rPA + CpG ODN had an ear thickness of 0.1519 (p &lt; 0.001 vs rPA), significantly greater than swelling induced by C 48/80 (p &lt; 0.001). Day 42 serum from mice immunized with PA alone had serum anti-PA IgG titers of 1:27,238 while mice immunized with PA + C 48/80 had anti-PA titers of 1:10,568,984 (p &lt; .001 vs PA alone). Similar titers were detected in mice vaccinated with rPA plus CpG ODN. Antibodies induced with rPA + C 48/80 neutralized anthrax lethal toxin. Our findings suggest that mast cell activators provide effective vaccine adjuvant activity when delivered intradermally while inducing less injection site inflammation than CpG ODN.
CD28 costimulation is required by effector and memory CD8+ T cells (43.2)
Authors: Boesteanu Alina C. , Dolfi Douglas V , Oliai Caspian H , Borowski Annie , Katsikis Peter D 
Published: January 2023

Abstract
CD28 costimulation is crucial for priming of CD8+ T cell responses to pathogens, however its role in the later phase of the primary and in the secondary immune responses is not clear. To dissect the role of CD28, C57Bl/6 mice were primed intranasally with influenza virus and then treated with a non-depleting blocking anti CD28 antibody at days 6 and 8 post infection. The frequency and absolute number of virus-specific CD8+ T cells were reduced in anti CD28 treated (3.9±0.4%; 1.9±0.5x105) vs. untreated mice (11.5±1.2%; 7.4±1.7 x105). We found that anti-CD28 treatment induces apoptosis of virus-specific CD8+ cells while it did not affect the number of actively dividing cells. To study the role of CD28 during secondary CD8+ T cell responses, C57Bl/6 mice were primed with flu virus strain PR8 and day 60 memory cells were transferred into C57Bl/6 or CD80−/−CD86−/− deficient mice. Mice were then rechallenged with flu virus strain X31 and 7 days post rechallenge CD80−/−CD86−/− deficient mice exhibited a reduction in virus-specific CD8+ T cells in the lung, compared to C57Bl/6 mice (1.2±0.7x106 vs. 5.8±1.1x106, respectively). Failure of memory CD8+ T cells to expand in the absence of CD28 costimulation is due to a failure to downregulate Bcl-2 and to cell cycle arrest. Thus, effector and memory CD8+ T cells require CD28 costimulation to generate optimal immune responses against pathogens.