bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inflammasomes/IL-1 pathways in myeloid cells modulate PD-1/PD-L1 checkpoint molecules
Authors: Guo Beichu , Zhang Jinyu , Fu Shunjun 
Published: January 2023

Abstract
The inflammatory microenvironment is not only critical for tumor growth and progression, but also associated with immunosuppression. Expression of immune checkpoint molecules, such as PD-L1, in tumor cells and innate immune cells has been linked to diminished anti-tumor response. However, the connection between inflammation and expression of checkpoint molecules in myeloid cells and tumor cells is poorly characterized. Our recent studies have demonstrated that the inflammasome and IL-1β pathway plays a critical role in promoting tumor growth and metastasis. Our data show that inflammasome activation promoted the infiltration of myeloid cells, such as myeloid-derived suppressor cells and tumor-associated macrophages, into tumor microenvironments. Interestingly, we found that inflammasome/IL-1 pathways induced expression of immune checkpoint molecule PD-L1 on infiltrated myeloid cells and tumor cells. Furthermore, blocking IL-1R with IL-1R antagonist (IL-Ra) or anti-IL-1R antibody inhibits tumor growth and metastasis accompanied by decreased accumulation of myeloid cells and expression of the PD-L1 molecule. Together, our results demonstrate that inflammasomes/IL-1 pathways promote infiltration of myeloid cells and expression of PD-L1, contributing to tumor immune evasion and tumor progression.
Defective STING signaling in ovarian cancer cells favor oncolytic virus action
Authors: de Queiroz Nina Marí Gual Pimenta , Xia Tianli , Barber Glen N. 
Published: January 2023

Abstract
Ovarian cancer is the sixth most prevalent cancer in women and the most lethal of the gynecologic malignancies. Our group has recently shown that STING signaling is often suppressed in a wide variety of cancers, including colorectal carcinoma and melanoma. In this work, we examined the expression and activation of STING pathway in 11 different human ovarian cancer cell lines (CA), generated from the diagnosis of ovarian cancer in varied stages and cultured in vitro. We evaluated STING expression by Western blot in these cell lines and showed that STING is present in 8 of the 11 cell lines. Signaling via STING requires the activation of cGAS synthase enzyme, which was absent in 7 cell lines. In the absence of both STING and cGAS all STING pathway was affected with no IRF-3 and NFκB-p65 translocation and activation. In order to correlate expression levels with STING function in the production of IFN-β, we transfected cells with dsDNA90 to activate STING or with Poly(I:C) for activation via RIG-I. ELISA and Real Time-PCR analyses showed that all 11 CA responded poorly to dsDNA90 stimulation. In contrast, majority of CA were able to produce IFN-β and CXCL10 in response to Poly(I:C), indicating that RIG-I pathway was preserved. Defective production of these cell death control cytokines after DNA stimulus may favor CA proliferation, but also facilitates viral replication and make this CA highly susceptible to HSV1γ34.5 (Herpes simplex virus). Analyzes in vivo by injection of human ovarian cancer cells subcutaneously in Nu/Nu mice followed by HSV1γ34.5 treatments correlates with in vitro findings. The results suggest that STING pathway can be suppressed during ovarian cancer development and these defects let CA more susceptible to oncolytic viral therapy.
Characterize the role of cancer associated fibroblast (CAFs) derived IL-32 in esophageal squamous cell carcinoma (ESCC)
Authors: CHEN Miao , GUAN Xin-yuan 
Published: January 2023

Abstract
Cancer-associated fibroblasts (CAFs) are localized adjacently to cancer cells at all stages of esophageal squamous cell carcinoma (ESCC) progression, and their functional contributions to this process are emerging. Our previous studies have identified FGFR2 as CAFs specific marker in ESCC. In this study, using microarray analysis, we identified IL-32 was the most differentially expressed cytokine in CAFs in patients with ESCC. Flow cytometric analysis and western blot have revealed markedly increased IL-32 in FGFR2+ CAFs from ESCC tumor specimens, compared with non-tumor epithelial tissues. IL-32 treatment could promote prosurvival effect in tumor cell from upon cisplatin or 5-Fu mediated apoptosis in culture and human ESCC tumor-bearing nude mice. In contrast, knockdown of IL-32 expression in CAFs by shRNA reduce tumor cell viability in reconstituted mouse model upon cisplatin or 5-Fu treatment. Mechanistic study shows that IL-32 treatment significantly downregulated caspase 3/9 and upregulated Bcl-2 expression by ESCC tumor upon Cisplatin or 5-Fu stimulation. In contrast, blockage of type II IFN signaling in CAFs resulted in significantly reduced IL-32 secretion and increased apoptotic cell death of ESCC. Together, these findings have suggested a critical role of CAFs derived IL-32 in prosurvival effect ESCC and thereby exacerbating cancer development, which suggested that IL-32 blockade may serve as potential therapeutic target for ESCC.
Rise in Interleukin-17-producing invariant Natural Killer T Cells during Bordetella Pertussis Infection in Mice
Authors: Santos Daniel Rebelo , Nguyen Alana , Bui Khoa , Abramson Tzvia 
Published: January 2023

Abstract
Bordetella pertussis is a Gram-negative bacterium known for causing the respiratory disease whooping cough. Whooping cough has recently seen resurgence, especially among children, with the disease’s resolution being delayed. Here, we observed the contribution of invariant Natural Killer T (iNKT) cells in the response to B. pertussis infections. Invariant NKT cells are so called for their invariant chain (Vα14Jα18 in mice) within the T cell receptor, which allows these cells to respond to lipid antigens presented by CD1d molecules. iNKT cells are known to express a variety of transcription factors and release different cytokines, as well as interact with numerous cells of the immune system. Here we hypothesize that IL-17 producing iNKT (iNKT17) cells rise in number during B. pertussis infections to aid in the resolution via an IL-17 driven inflammation, as well as to activate antigen presenting cells. The population of iNKT17 cells was assessed via comparison of uninfected to B. pertussis-infected C57BL/6 mice, using a multicolor flow cytometry analysis. The mice infected with pertussis displayed a significant rise in iNKT cells when compared to uninfected mice, while being positive for RORγt and IL-17 secretion. Immunohistochemical analysis displays the interaction of iNKT cells with resident dendritic cells during infection. These findings indicate a role for iNKT17 cells in the pathogenesis and potential resolution of B. pertussis infection, with the potential for future analysis into iNKT cell knockout studies to determine further understanding of the cell’s function during the infection.
Early-life respiratory bacterial infection-induced chronic lung disease is driven by a novel TLR2/IL-13/miR-21/PI3K-dependent, but MyD88-independent signalling pathway
Authors: Starkey Malcolm Ronald , Nguyen Duc H , Kim Richard Y , Nair Prema M , Haw TattJhong , Horvat Jay C , Godfrey Dale I , McKenzie Andrew N , Hansbro Philip M 
Published: January 2023

Abstract
There is a critical window in early-life where the lung is still maturing and susceptible to infection. Indeed, severe respiratory infections in early-life are a risk factor for the development of chronic lung diseases. The aim of this study was to identify the mechanisms involved.
Neonatal wild-type (WT), TLR2 deficient (−/−), IL-13−/−, MyD88−/− and STAT6−/− mice were infected with the natural mouse bacterial pathogen Chlamydia muridarum, as a model of severe respiratory tract infection in early-life. In some experiments WT mice were treated with miR-21 antagomirs, PI3K inhibitors, or relevant controls during early-life infection. The impact of targeting these specific immune molecules during early-life on infection-induced impairment of lung function and structure were assessed.
Neonatal Chlamydia respiratory infection increased TLR2, IL-13-receptor, miR-21 and PI3K expression and/or activity in the lung. TLR2 signalling induced IL-13-receptor expression, IL-13 signalling induced miR-21 expression and miR-21 increased PI3K activity. TLR2 signalling also increased IL-13+ ILC2s in the lung. TLR2−/− and IL-13−/− mice were protected against infection-induced airway hyperresponsiveness (AHR), but not emphysema-like alveolar enlargement. This TLR2/IL-13 mediated phenotype was independent of MyD88, but dependent on STAT6. Specific targeting of miR-21 prevented AHR but not emphysema. Pan-PI3K inhibition did not affect AHR, but protected against emphysema. Interestingly, early-life infection-induced AHR was steroid insensitive.
This study identifies a novel signalling network that may be targeted for the prevention of the long-term deleterious effects of early-life infection on lung function and structure.
Differential regulation of development of experimental autoimmune encephalomyelitis by Akt-1 and Akt-2
Authors: Zeng Qiuming , Ouyang Song , Guo Hui , Tang Rong , Liu Jishi , Zhang Jian 
Published: January 2023

Abstract
Akt-1 and -2 are major members of serine/threonine Akt family kinases that play a key role in multiple cellular processes. Previously, we have shown that Akt-2 but not Akt-1 is involved in the inhibition of inducible regulatory T cell development. However, the role of both isoforms of Akt in the development of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, is completely unknown. In this study, we showed that while Akt1−/− mice develop ameliorated EAE, Akt2−/− mice develop severe disease. At the cellular levels, Akt-1 appears to inhibit proliferation of thymic-derived CD4+CD25+Foxp3+ regulatory T cells (tTregs), but facilitates antigen-specific TH1/TH17 responses during EAE induction. In a sharp contrast to Akt-1, Akt-2 potentiates tTreg proliferation, and suppresses antigen-specific TH1/TH17 responses. Our data demonstrate Akt-1 and Akt-2 differentially regulates the susceptibility of mice to EAE possibly by controlling tTreg proliferation in the periphery, thus controlling TH1/TH17 responses during EAE development.
Plasma cells and not memory cells, constitute the majority of human IgE Cells
Authors: Saunders Sean P , Narayanan Sriram , Narang Viping N. , Wong Glenn , Aw Derrick C.W. , Chandran Nisha , Navin Ishwar , Reibman Joan , Poidinger Michael , Zolezzi Francesca , Curotto De Lafaille Maria A. 
Published: January 2023

Abstract
Immunoglobulin E (IgE) is central in allergy pathology as it binds high-affinity receptors on mast cells, inducing degranulation upon allergen crosslinking. However, IgE has the lowest serum concentration and shortest half-life of all Ig’s, and IgE+ B cells are extremely rare. This work aims to uncover characteristics of IgE+B cells that could be used in the design of new therapeutic strategies for allergic diseases.
IgE+ cells were identified in Atopic Dermatitis (AD) patients, and were also generated in vitro by stimulating with aCD40 and IL-4. Naïve, IgG+, IgA+ and IgE+ cells from AD donors were phenotyped and sorted by flow cytometry, and their transcriptomes were determined by RNAseq. Inter-sample similarity was determined using multidimensional scaling (MDS). Gene ontology (GO) of differentially expressed genes (DEG) identified molecular pathways enriched in IgE+ cells.
IgE+ cells were CD38hiCD27hi, indicating a high content of plasma cells in IgE+ B cells. IgE+ cells expressed high levels of proliferation marker Ki67. MDS comparison of datasets of naïve, IgG+, IgA+ and IgE+ cells showed that IgE cells were distant from both IgG and IgA cells. GO analysis of IgE DEG demonstrated that IgE cells expressed genes associated with N-glycosylation, the IRE1-XBP1 endoplasmic reticulum unfolded protein response, and mitochondrial oxidative phosphorylation, synonymous with plasma cells.
Our data indicates that IgE+ cells in AD patients are predominantly plasmablasts and plasma cells, and not memory B cells. Ongoing studies aim to characterize human IgE cell diversity using single cell analysis, and to identify non-IgE precursor memory cells that give rise to IgE+ cells upon allergen activation.
Impact of Nur77 on the initiation and development of E. coli induced acute lung injury
Authors: Hou Jinchao 
Published: January 2023

Abstract

Objectives
Orphan nuclear receptor Nur77 (Nur77) has been emerged as a regulator of gene expression in aseptic inflammation, but its role in regulating macrophage response to bacterial infection remains unclear. So, this study was designed to investigate the impact of Nur77 in E. coli induced acute lung injury.

Methods
Mice with different genotypes (WT, Nur77−/−) were challenged with E. coli intratracheal inoculation. Mortality was assessed hourly. Bronchoalveolar lavage fluid (BALF) and blood were obtained and bacterial burdens were assayed using agar plating method. Lung morphology changes and pulmonary vascular permeability were assessed by HE staining or BALF protein accumulation, separately. The TNF-α level in the BALF were determined by ELISA. Ex vivo, the Nur77 mRNA level expressed on the alveolar macrophages (AMs) under rest or E. coli stimulation were detected by qPCR. The phagocytosis ability of AMs from WT and Nur77−/− mice were further assessed.

Results
The survival rate of the Nur77−/− mice was 93.3%, while only 31.3% of WT mice survived more than 48 hours. Bacterial counts in the BALF collected from the WT mice were higher than those from the Nur77−/− mice by 10 folds. Along with reduced pulmonary bacterial load, diminished lung injury and permeability were observed in the Nur77−/− group. In addition, we found largely decreased TNF-α level in Nur77−/− mice compared with WT mice in BALF. The mRNA level of Nur77 was largely up-regulated in AMs under E. coli stimulation ex vivo. Functionally, Nur77−/− AMs exhibited increased rates of bacterial engulfment.

Conclusions
The absence of Nur77 enhances lung immune defense to eliminate invading E. coli and prevents the pathogen from spreading, ultimately improving the outcome.
Human neutrophils alter IL-20 receptor expression and IL-20 modulates human neutrophil function during in vitro infection with Staphylococcus aureus
Authors: Datta Sandip K , Gough Portia 
Published: January 2023

Abstract
Neutrophils are critical to a protective immune response to infection with Staphylococcus aureus, a leading cause of skin and soft tissue infections. The responses of neutrophils are shaped by environmental cues in the form of the type of pathogen encountered and host signals in the form of cytokines and chemokines. Yet, there is an incomplete understanding of signaling at the site of infection and how signals work in concert to influence neutrophil function and the biological tug-of-war that occurs between host and pathogen. Previous work showed that IL-20 is produced by human keratinocytes infected with S. aureus in vitro, and during a subcutaneous model of infection in mice in vivo. Signaling through an IL-20 receptor reduced neutrophil recruitment via reduced levels of IL-1β and IL-17A, thereby contributing to the pathogenesis of infection. Since IL-20 is produced by infected epithelial cells, we investigated if this cytokine could affect neutrophils directly. We found that neutrophils alter IL-20 receptor expression in response to in vitro infection with S. aureus and exposure to pro-inflammatory host factors, TNFα and IL-8. In these conditions, IL-20 altered actin dynamics in neutrophils and affected functions that rely on this activity: migration, phagocytic uptake and tertiary granule exocytosis.
Control of regulatory T (Treg) cell function by protein kinase C-eta (PKCη): A novel target for cancer immunotherapy
Authors: Pedros Christophe , Kong Kok-Fai , Altman Amnon 
Published: January 2023

Abstract
Modulating regulatory T cell (Treg) function to promote anti-tumor immunity is a desirable strategy for cancer immunotherapy. Blockade of checkpoint molecules CTLA-4 and PD-1 has shown clinical efficacy in metastatic melanoma and other malignancies. We recently reported that Treg stimulation induced the recruitment of protein kinase C-eta (PKCη), together with the GIT-PAK-PIX signaling complex that mediates focal adhesion disassembly, to CTLA-4, an inhibitory receptor highly expressed on Foxp3+ Tregs and important for Treg suppression. As a result, PKCη was recruited to the immunological synapse of stimulated Treg. These events promoted Treg-mediated contact-dependent suppression, including suppression of tumor immunity, via a mechanism likely involving depletion of costimulatory CD80/86 ligands from APCs (Nat Immunol. 2014;15:465–72).
PKCη-deficient (PKCη−/−) Tregs were defective in the suppression of anti-tumor immunity against B16 melanoma and the prostate adenocarcinoma TRAMP-C1 but retained the ability to inhibit experimental colitis, a disease that often occurs in patients treated with anti-CTLA-4 antibodies. PKCη loss in mouse Treg cells increased both the number of intratumoral CD4+ and CD8+ effector T cells and their ability to produce cytokines upon restimulation. Moreover, shRNA-mediated knockdown of PKCη reduced the suppressive activity of human Tregs. Impaired activation of the GIT-PAK-PIX complex and the resulting defect in CD80/86 depletion are likely responsible for the defective suppressive activity of PKCη−/− Tregs. Altogether, our findings establish the CTLA-4-PKCη signaling pathway as critical for suppressing tumor immunity, and implicate it as an attractive cancer immunotherapy target.
Synergistic effect of IFN-γ on breast cancer targeted therapy
Authors: Nagai Yasuhiro , Tsuchiya Hiromichi , Ji Mei Q , Zhang Hongtao , Greene Mark I 
Published: January 2023

Abstract
ErbB2 is amplified in ~30% of breast cancer patients, and its amplification is associated with poor prognosis and worse survival outcome. We examined the synergistic effect of interferon-γ (IFN-γ) with anti-erbB2/neu mAb on erbB2-positive cancer model. IFN-γ showed marginal effects on its own, indicating that immune therapies mediated by this cytokine alone are unlikely to be beneficial. However, we discovered that treatment of the tumors with anti-erbB2/neu mAb concomitant with IFN-γ led to dramatic inhibition of in vivo tumor growth in the syngeneic tumor model and the preventional MMTV-neu transgenic mouse tumor model. We noted an increase of M1 macrophage accumulation in the tumor tissues as well as diminished myeloid derived suppressor cells, and increased CD8+ T cell cytotoxicity against tumor cells in the IFN-γ plus anti-erbB2/neu mAb treated mice. The tumor cells treated with both mAb and IFN-γ underwent changes in tumor stem cell marker ALDH-1, indicating a loss of stem cell-like properties, while mAb treatment alone did not accomplish this phenotypic change. We found that IFN-γ treatment significantly increase tumor PD-L1 expression, so we included anti-PD-1 or PD-L1 antibody with this combination therapy. Interestingly, anti-PD-L1 antibody further reduced tumor growth, while anti-PD-1 did not affect very much. These results indicate that the treatment of IFN-γ can improve targeted therapy, and further induce PD-L1 expression on tumor cells, creating a new target on the tumor cells for anti-PD-L1 therapy.
Identification of potential immune targets in controlling Endometrioid Endometrial Carcinoma metastatic progression.
Authors: Billaud Jean-Noel , Tugendreich Stuart , Toburen Debra 
Published: January 2023

Abstract
Endometrial adenocarcinoma is a common cause of gynecological cancer death in North America. The most dominant subtype, Endometrioid Endometrial Cancer (EEC) accounts for &gt;80% of this cancer and is estrogen-dependent. At diagnosis, 75% of women have the disease confined to the uterus, which is considered Stage One. Five-year survival for Stage One patients is 80%, however, about 15–20% develop metastasis.
Total RNA extracted from tissues obtained after surgical resection from three women at stage one EEC was subjected to RNA-sequencing. The publicly available dataset (SRP045645) was downloaded directly from the Sequence Read Archive and the FASTQ files were processed with Biomedical Genomics Workbench for secondary analysis including mapping, quantification and differential expression analysis. Through streamlined integration the data was uploaded to Ingenuity Pathway Analysis (IPA) for biological interpretation.
We show how in silico solutions developed by QIAGEN Bioinformatics enable us to analyze the biological wparameters involved in EEC metastatic progression from an early stage. By comparing these patients’ RNA-seq results, we determined that key Canonical Pathways and other biological processes differentiated the three patients from one other. The predicted transcriptional program allowed us to visualize key upstream drivers, three of these drivers are GDF15, FGF3 and EDN1. They have been predicted to be potential master regulators of a Causal Network that increase metastasis, epithelial-to-mesenchymal-transition, and cellular invasion in one of the three patients. Based on these results we propose these three immune molecules as possible therapeutic targets to counteract the metastatic processes in EEC
Extensional stress modulates leukocyte adhesion probability
Authors: Benson Bryan , Ransohoff Richard Milton , Huang Alex Yee-Chen 
Published: January 2023

Abstract
Post capillary venules (PCVs) contribute a tiny fraction of the total vasculature, but are the predominant site of leukocyte adhesion and infiltration. Given the astronomical number of leukocytes and length of the vasculature, this restriction of adhesion to PCV requires tight and redundant regulation: adhesion molecule expression, chemokine presentation, and wall shear stress (WSS) from blood flow converge to promote this phenomenon in PCVs.
To mimic WSS, parallel plate fluidic channels have become the standard of in vitro models to study leukocyte adhesion. These models match the WSS of in vivo PCVs, but vary in their height (h), the smallest dimension that most strongly dictates fluid forces. Surprisingly, in our own experiments and in literature, leukocyte adhesion decreases as h decreases towards the 25 micron diameter of in vivo PCVs, whereas computational studies predict more adhesion with smaller h. This discrepancy suggests that a biological response within leukocytes opposes passive physical forces, or that other mediating factors are at play in vivo.
In the transition from capillary to PCV, leukocytes experience another fluid stress besides WSS: extensional stress, due to the cross-sectional area of the vessels increasing at bifurcations.
We created biomimetic models of the capillary to PCV transition in which we independently vary extensional stress and WSS and assay leukocyte adhesion to adsorbed CXCL12 and ICAM-1. Here, extensional stress has a powerful influence on human peripheral blood mononuclear cell (PBMC) and Jurkat adhesion probability, with higher extensional stress strongly promoting leukocyte adhesion. We are currently investigating the roles of mechanosensitive channels PIEZO1 and TRPV2 in this effect.