bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pre-clinical efficacy of dual PI3K δ/γ inhibitors in airway inflammation (64.2)
Authors: Nagarathnam Dhanapalan , Govindarajulu Babu , Routhu Kasiviswanath , Rajagopalan Prasanna , Veeraraghavan Sridhar , Viswanadha Srikant , Vakkalanka Swaroop 
Published: January 2023

Abstract
Dual δ/γ inhibition is strongly implicated as an intervention treatment in allergic and non-allergic inflammation of the airways and autoimmune diseases. The objective of this study was to explore the therapeutic potential of RP6047, a novel, small molecule PI3Kδ/γ inhibitor in airway disorders. Specificity of lead molecule, RP6047, towards PI3Kδ and γ isoforms and subsequent downstream activity was determined in enzyme and several cell based assays. Pre-clinical efficacy of RP6047 was confirmed in animal models of airway inflammation. RP6047 demonstrated significant potency against PI3Kδ (19.7 nM) and γ (31.2 nM) with several fold selectivity over α and β isoforms and subsequent pAKT inhibition. Additionally, the compound inhibited neutrophil functionality and mast-cell degranulation at nanomolar concentrations besides reversing CSE induced dexamethasone insensitivity by &gt;90%. RP6047 displayed excellent efficacy in inhibiting LPS-induced TNFα (36% at 10 mg/kg/po) and neutrophilia (ED50 = 1.35 mg/kg/po) in a rat model. Administration of compound also resulted in a near complete reduction in inflammatory cell infiltration into the lungs in CSE treated mice over a 4-day period. Results demonstrate the therapeutic potential of RP6047 in asthma and related airway disorders acting via the PI3Kδ/γ pathway. Further in vivo studies are in progress to evaluate the efficacy of the compound in airway hyper-responsiveness prior to the initiation of clinical trials.
Neutrophils: key players in mast cell-mediated thermal pain in mice (67.4)
Authors: Chatterjea Devavani , Willenbring Robin , Balsells Evelyn , Paredes Luisa , Allen Juliann 
Published: January 2023

Abstract
Mast cells are involved in a range of physiologies and pathologies including allergic disease, defense against pathogens, and neutralization of venoms. Recently, these cells have been shown to play significant roles in inflammatory pain. Acute inflammation is a beneficial mechanism designed to prevent further tissue damage; however, it can also lead to a decreased quality of life and, as such, is a pervasive health concern. The involvement of mast cells in this process has not been fully characterized. We show, in a mast cell-dependent model of thermal pain that degranulation of peripheral mast cells in the plantar tissue of mice is accompanied by neutrophil influx as well as edema and upregulation of pro-inflammatory cytokines. When mast cells are stabilized with sodium cromoglycate, pain, edema and neutrophil influx are abrogated. Additionally, when neutrophil influx is blocked with pre-administration of the selectin blocker fucoidan, pain and edema are also abrogated. This suggests neutrophil influx is a key event in the development of mast cell-initiated inflammatory pain.
A novel autocrine-paracrine loop of TSLP in atopic dermatitis is modulated by p63. (71.4)
Authors: Kubo Terufumi , Ye Rui , Ara Shihoko , Matsumiya Hiroshi , Kojima Takashi , Kikuchi Tomoki , Sato Noriyuki , Ichimiya Shingo 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a common disease affecting over 10% of children and adults, which severely impairs quality of life. Accumulating evidence has revealed that thymic stromal lymphopoietin (TSLP) of a pleiotropic cytokine play a critical role in the maintenance of AD. Currently, epidermal cells of AD are postulated as a major producer of TSLP, eventually leading to unfavorable local inflammation. However a property of the epidermal cells of AD is largely unknown. In the present study we assessed a functional significance of an epitheliotropic transcription factor p63 of a p53 homologue in the skin lesions of AD. Interestingly immunohistochemical analysis indicated constitutive downregulation of p63 of the epidermal cells in AD-affected skin areas. Further in vitro experiments using primary epidermal cells demonstrated that TSLP stimulation efficiently allowed cells to secrete TSLP as well as several inflammatory cytokines. Moreover it was found that the levels of expression of TSLP receptor (TSLPR) were under the control of p63. Collectively these indicate that the downregulation of p63 may cause chronic inflammation through providing a harmful TSLP-TSLPR axis to epidermal cells. Together with the evidence that innate ligands to TLRs like TLR3 could steadily suppress p63, our data would provide new insight into the pathogenesis of AD, especially with respect to the operation of innate immune responses by epidermal p63.
Ligation of CTLA-4 to the TCR inhibits T cell activation and directs differentiation into Foxp3+ regulatory T cells (72.2)
Authors: Zhu Yunxiang , Karman Jozsef , Jiang Ji-Lei , Gumlaw Nathan , Zhao Hongmei , Campos-Rivera Juanita , Sancho Jose , Zhang Jinhua , Jiang Canwen , Cheng Seng H. 
Published: January 2023

Abstract
ABSTRACT Crosslinking of ligand-engaged cytotoxic T lymphocyte antigen-4 (CTLA-4) to the T cell receptor (TCR) during the early phase of T cell activation attenuates TCR signaling, leading to T cell inhibition. To promote this event, a bispecific fusion protein comprising a mutant mouse CD80 (CD80w88a) and lymphocyte activation antigen-3 (LAG-3) was engineered to concurrently engage CTLA-4 and crosslink it to the TCR. Crosslinking is expected to attain via ligation of CTLA-4 first to MHC II and then indirectly to the TCR, generating a CTLA-4-MHC II-TCR tri-molecular complex that forms between T cells and antigen-presenting cells (APCs) during T cell activation. Treating T cells with this bispecific fusion protein inhibited T cell activation. In addition, it induced the production of IL-10 and TGF- β and attenuated AKT and mTOR signaling. Intriguingly, treatment with the bispecific fusion protein also directed early T cell differentiation into Foxp3+ regulatory T cells (Tregs). This process was dependent on the endogenous production of TGF- β. Thus, bispecific fusion proteins that engage CTLA-4 and co-ligate it to the TCR during the early phase of T cell activation can negatively regulate the T cell response. Bispecific biologics with such dual functions may therefore represent a novel class of therapeutics for immune modulation. The findings presented here also reveal a potential new role for CTLA-4 in Treg differentiation.
Control of IL-2/ regulatory T cell axis by Calcium/calmodulin-dependent protein kinase type IV plays a key role in lupus-prone mice (72.4)
Authors: Koga Tomohiro , Ichinose Kunihiro , Mizui Masayuki , Crispín José , Tsokos George 
Published: January 2023

Abstract
Objective. Dysfunction of regulatory CD4+CD25+FoxP3+ T (Treg) cells has been detected in autoimmune diseases. IL-2 is critical for the development, survival, and function of Treg cells. Increased levels of Calcium/calmodulin-dependent protein kinase type IV (CaMKIV) cause defective IL-2 production in T cells from patients with SLE. Methods. We crossed CaMKIV deficient (CaMK4-/-) mice with MRL/MpJ-lpr/lpr (MRL/lpr) mice and examined survival rate, autoantibody levels, immune cells subpopulations, pro-inflammatory cytokine production and T cell function. Results. The survival rate was significantly higher in MRL/lpr.CaMK4-/- mice than in wild type MRL/lpr mice at 32 weeks (89% versus 25%, p&lt;0.05). MRL/lpr.CaMK4-/- mice displayed decreased titers of anti-DNA antibodies, reduced numbers of double negative T cell, CD4+ activated/memory cells in the spleen. The production of IL-2 in response to anti-CD3/28 antibody by CD4+ T cells was restored in MRL/lpr.CaMK4-/- mice, and this was associated with decreased levels of phosphorylated cAMP response element modulator (CREM) in the nucleus. Notably, the number of Treg cells was increased in MRL/lpr.CaMKiv-/- mice. We further confirmed silencing of CaMKIV resulted in overexpression of FoxP3 gene in human SLE T cells. Conclusion. Our results suggest that CaMKIV is important in the expression of the disease in lupus-prone mice, and we propose that CaMKIV represents a valuable target for treatment in autoimmune diseases.
The role of TRIM68 in Toll-like receptor and RIG-I-like receptor induced interferon production (72.5)
Authors: Wynne Claire , Higgs Rowan , Biron Christine , Jefferies Caroline 
Published: January 2023

Abstract
Innate immune receptors such as Toll-like receptors (TLR) and RIG-I-like receptors (RLR) play key roles in viral recognition and interferon (IFN) production. Systemic Lupus Erythematosus (SLE) patients commonly present with an over production of type 1 IFNs which contribute to disease pathogenesis so understanding how to dampen down this response is of huge importance. The focus of this project is on the E3 ubiquitin ligase TRIM68, a known autoantigen in SLE. Results show TRIM68 to be a novel negative regulator of type 1 IFN production in both TLR- and RLR-dependent pathways. Proteomic analysis of TRIM68 containing complexes has allowed us to identify TRK-fused gene (TFG) as a TRIM68 target. The physiological role of TFG is still unknown but it has been shown to interact with NEMO and TANK, two proteins which modulate pro-inflammatory cytokine production. Thus the effects of TRIM68 on IFN production is via this novel regulation of TFG. To confirm the critical role of TRIM68 in IFN production we have knocked down TRIM68 in mice via injection of PLGA microparticles encapsulating shRNA against TRIM68 and studied the effects of TRIM68 knock down on viral-induced IFN production. TRIM68 knockdown mice display an increase in serum and peritoneum IFN levels post LCMV infection, indicating that TRIM68 is a key negative regulator of IFN production. This work carves the way for the development of novel therapeutics targeting TRIM68 for the treatment of IFN-driven disease such as SLE.
Expression of the tumor-associated antigen Neu5Gc-Sialyl-Tn in human carcinomas (74.6)
Authors: Padler-Karavani Vered , Deng Liwen , Garcia Andrea , Karp Felix , Naidu Natasha , Choudhury Biswa , Varki Nissi , Varki Ajit 
Published: January 2023

Abstract
Altered glycosylation is prevalent on cancer cells, due to aberrant expression of glycosyltransferases, and frequently involving variations in sialylation patterns. In humans, this altered sialylation can also result from metabolic incorporation of dietary N-glycolylneuraminic acid (Neu5Gc) that differs from the human sialic acid N-acetylneuraminic acid (Neu5Ac) by one oxygen atom. Neu5Gc is immunogenic in humans and anti-Neu5Gc antibodies can either promote tumor growth at low doses or cause tumor regression when administered in excess. Antibodies against Neu5Gcα2−6GalNAcα1-O-Ser/Thr (anti-Neu5Gc-Sialyl-Tn IgG) appear to be novel carcinoma biomarkers, but direct evidence for expression of this epitope on human carcinomas is lacking. This neo-sialoglycan resembles the well-known carcinoma-associated biomarker Sialyl-Tn, except that Neu5Ac is replaced with dietary Neu5Gc. Here, we characterize unique antibodies against Neu5Gc and Sialyl-Tn by sialoglycan-microarrays and use them for immunohistochemistry of human breast, prostate, ovary, colon and lung carcinomas revealing co-staining in some mucinous carcinomas. We also generate MUC1-Fc that is heavily glycosylated with Neu5Gc-Sialyl-Tn for affinity-purification of human anti-Neu5Gc-Sialyl-Tn-MUC1 IgG for further immunohistochemistry analysis. Finally, we report a mass spectrometry approach to analyze O-linked glycans from tissues to ultimately prove biomarker expression in positively-stained tissues.
The mechanisms of photodynamic therapy enhancement of anti-tumor immunity (75.1)
Authors: Wang Minan , Gollnick Sandra 
Published: January 2023

Abstract
Photodynamic therapy (PDT) has long been shown to be capable of killing malignant cells and inducing host immune response. Recent investigations have also established that PDT augments anti-tumor immunity. Tumor cells treated by PDT release “danger signals” that have been shown to stimulate dendritic cells (DCs). In this study, we investigate the role of danger signal receptors, specifically the toll-like receptors (TLR) and the NOD-like receptor NLRP3, in PDT generated tumor cell lysates activation of DCs. Stimulation of bone marrow derived DCs (BMDCs) with PDT treated tumor cells leads to dramatic increase in pro-inflammatory cytokine production, including IL-1β, which plays a critical role in PDT efficacy. Production and activation of IL-1β require both TLR and NLR signaling. IL-1β production induced by PDT vaccine in BMDCs is MyD88 dependent. NF-κB is also activated upon treatment of BMDCs with PDT treated tumor cell lysates. Caspase-1, an important component of NLRP3 inflammasome, is known for its role in maturation of IL-1β. However, we found that IL-1β release from DCs is caspase-1 independent. This study aims to understand how PDT enhances tumor immunogenicity and provides insight to improve both in situ PDT and PDT vaccine efficacy in treating secondary disease.
Targeting the OX40 (CD134) co-stimulatory receptor enhances mucosal secondary memory CD8 T cells (P270) (75.7)
Authors: Goulding John , Croft Michael , Salek-Ardakani Shahram 
Published: January 2023

Abstract
The expansion and maintenance of poly-functional memory CD8 T cells (mCD8) that rapidly migrate to sites of viral replication is paramount for the development of effective mucosal vaccines. This concept is epitomized by the limited success of vaccines that utilize attenuated or replication defective virus strains incapable of promoting mCD8 T cells that persist at mucosal sites. Consequently, strategies that enhance both the number and functionality of mCD8 T cells have attracted increasing interest. Recent studies have shown that targeting of the OX40 co-stimulatory receptor can promote primary mucosal CD8 T cell memory following infection with virulent and attenuated pox-virus vaccine strains. However, the ability of OX40 co-stimulation to enhance the reactivation of secondary mCD8 T cells is unclear. To mimic a prime boost vaccination strategy we transferred virus specific mCD8 T cells into naïve mice that were later immunized via dermal scarification. We show that OX40 agonism 24 hours following vaccination enhances both the number of virus specific secondary effector mCD8 T cells, defined by their expression of KLRG, IL7-R, CD62L and CD27, located within the lung and their capacity to produce both TNFα and IFNγ. These data provide evidence that targeting OX40 at the same time as an antigen boost (secondary challenge) can expand the numbers of effector mCD8 T cells that are capable of residing within lung tissue and thus protect against mucosal viral infection.
Presentation of cryptic peptides by MHC I molecules is enhanced by inflammatory stimuli. (P5003)
Authors: Prasad Sharanya , Starck Shelley , Shastri Nilabh 
Published: January 2023

Abstract
Cytolytic T cells eliminate infected cells by recognizing intracellular peptides presented by MHC class I molecules. The antigenic peptides are derived primarily from newly synthesized proteins including those produced by cryptic translation. Previous studies have shown that in addition to the canonical AUG codon, translation can be initiated at non-AUG codons . Furthermore, translation initiation at non-AUG codons such as CUG is mechanistically distinct from canonical translation initiation as it is resistant to protein synthesis inhibitors that cause global translation shutdown. Here, we show that Toll-like receptor (TLR) signaling pathways involved in pathogen recognition enhance presentation of the cryptic peptides. Moreover, infection of bone-marrow derived macrophages with MCMV, influenza viruses or pro-inflammatory cytokines also enhances cryptic peptide presentation. Thus, translation and presentation of cryptic peptides may allow the immune system to detect intracellular pathogens that inhibit host translation and presentation of peptides from conventional sources.
IgE enhances B cell-derived exosomal induced T cell proliferation. (P5013)
Authors: Martin Rebecca , Brooks Keith , Henningson Frida , Heyman Brigitta , Conrad Daniel 
Published: January 2023

Abstract
The immunostimulatory potential of IgE immune complexes is known to be dependent on CD23, the low affinity IgE receptor. This was originally thought to be due to CD23 mediated uptake and subsequent stimulation of T cells by B cell processed antigen. Recent findings have indicated that dendritic cells (DC) are required in this process and B cells are primarily needed for antigen transportation. This has called into question the role of B cells in this interaction and leaves a gap in the knowledge of how the antigen is transferred to the DC. Given the resurgent interest in exosomes as having the capacity to mediate antigen presentation, we tested whether B cell derived exosomes might provide an explanation for this transfer. B cell derived exosomes express CD23 as well as the costimulatory molecules CD80 and CD86. Exosomes isolated from B cells that had been cultured with anti-TNP-IgE and TNP-OVA were shown to induce proliferation of ova specific DO11 T cells in a dose dependent manner. In the absence of either CD23 or anti-TNP-IgE, specific T cell stimulation was severely reduced. Thus, B cell derived exosomes alone have the capacity to induce antigen specific T cell activation and could represent the mechanism for antigen transfer to DCs. Ongoing testing involves whether injection of exosomes with processed antigen, either iv or intrasplenic will give an enhanced T cell and anti-OVA response in a manner similar to the classical IgE immune complex injection.
Regulation of gene expression by the immunoproteasome (P5049)
Authors: Rouette Alexandre , de Verteuil Danielle , Khanafer Hassan , Hardy Marie-Pierre , Perreault Claude 
Published: January 2023

Abstract
Background: Proteasomes play crucial roles in regulating fundamental cellular processes in eukaryotic cells. Catalytic subunits β1, β2 and β5 of the proteasome are replaced by LMP2, MECL-1 and LMP7 to form the immunoproteasome (IPr). We have shown that the IPr modulates mRNA levels of several clusters of genes in dendritic cells, but the precise mechanism by which it regulates gene expression is unknown. Results: By comparing transcriptome of Lmp7+/+/Mecl1+/+ (WT) and Lmp7-/-/Mecl1-/- (dKO) thymocytes by microarray, we found that the IPr affects the expression of 50 transcripts. The genes source of these transcripts are clustered in the genome, and are enriched in cell cycle-associated functions. Moreover, an overlap of 2% is seen by comparing with transcripts modulated by the IPr in dendritic cells. Chymotrypsin-like activity is higher in dKO than in WT cells, due to β5 overexpression. Furthermore, ubiquitinated proteins levels are similar between WT and dKO cells whereas free ubiquitin is lower in dKO cells. We next investigated levels of ubiquitin on histones, since it represent an important reservoir of ubiquitin and it can regulate transcriptional activity. Our results show that H2B, but not H2A, ubiquitination is increased in dKO cells. Conclusion: The effect of IPr on transcription is tissue-specific and could be due to increased ubiquitinated H2B levels. While proteasomes clearly regulate transcription, the impact of IPr on transcription has never been investigated.
Butyrophilin 3A1 plays an essential role in prenyl pyrophosphate recognition by human Vγ2Vδ2 T cells (P5051)
Authors: Morita Craig , Henry Olivier , Distefano Mark , Mönkkönen Jukka , Tanaka Yoshimasa , Wang Hong 
Published: January 2023

Abstract
The majority of human γδ T cells express Vγ2Vδ2 TCRs and play important roles in microbial and tumor immunity. Vγ2Vδ2 T cells use their TCRs to recognize self and foreign prenyl pyrophosphate intermediates (prenyl-PP) in isoprenoid synthesis. However, little is known about the molecular basis for this recognition. We find that a monoclonal antibody specific for butyrophillin 3 (BTN3) Ig superfamily proteins mimics prenyl-PPs. The 20.1 mAb stimulated Vγ2Vδ2 T cell clones regardless of their functional phenotype or developmental origin, and selectively expanded blood Vγ2Vδ2 T cells. The γδ TCR mediates 20.1 stimulation since IL-2 is released by β− Jurkat cells transfected with Vγ2Vδ2 TCRs. 20.1 stimulation was not due isopentenyl pyrophosphate (IPP) accumulation because 20.1 treatment of APC did not increase IPP levels. Nor was stimulation inhibited by statin treatment which blocks IPP production. Importantly, siRNA knockdown of BTN3A1 abolished stimulation by IPP that could be restored by re-expressing BTN3A1. Rhesus monkey and baboon APC presented IPP and 20.1 to human Vγ2Vδ2 T cells despite amino acid differences localized to the outer BTN3A1 face suggesting that the top and/or the inner face interact with its co-receptor. Full length BTN3A1 did not preferentially bind a photoaffinity prenyl-PP analog. Our results confirm and extend recent studies and demonstrate that BTN3A1 is required for the recognition of prenyl-PP but does not appear to bind the intermediates directly.
Heat shock protein 90 inhibitor radicicol attenuates matrix metalloproteinase expression in human monocytic cells (P4153)
Authors: Hsiao George , Lee Tzong-Huei , Jan Jing-Shium , Hsiao Che-Jen , Cheng Yu-Wen 
Published: January 2023

Abstract
The molecular chaperone heat shock protein 90 (HSP90) has emerged as an exciting target for anti-tumor and immunoregulatory therapy. In this study, we investigated the inhibitory effects and mechanism of radicicol, as a HSP90 inhibitor, on human monocytic matrix metalloproteinases (MMPs) and pro-inflammatory cytokine expression. Radicicol exhibited concentration-dependent inhibition of MMP-9 enzymatic activation induced by either tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS) in human monocytic THP-1 cells. Western blot and RT-PCR analyses showed that radicicol concentration-dependently suppressed LPS-induced monocytic MMP-9 protein and mRNA expression. It was also shown that radicicol enhanced HSP70 expression. Besides, we clearly found that radicicol restore the degradation of inhibitor-kappaB-alpha and abrogate NF-kappaB reporter gene expression in activated THP-1 cells. In addition, activation of Akt and its total protein expression were reduced by radicicol. Furthermore, radicicol presented an inhibitory effect on both TNF-alpha and interleukin-6 productions by ELISA. In conclusion, we demonstrate that radicicol might involve interference with the NF-kappaB or Akt signaling pathway to attenuate production of MMP-9 and pro-inflammatory cytokines. These results provide a new opportunity for the development of anti-leukemia and cachexia-relief strategies.