scholarly journals Stromal fibroblast activation and inflammation in frozen shoulder

2018 ◽  
Author(s):  
Moeed Akbar ◽  
Michael McLean ◽  
Emma Garcia-Melchor ◽  
Lindsay AN Crowe ◽  
Paul McMillan ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Immune Cell ◽  
Frozen Shoulder ◽  
Stromal Fibroblast ◽  
Stromal Fibroblasts ◽  
Activation Markers ◽  
Stromal Compartment ◽  
Activation Marker ◽  
Stromal Activation

AbstractIntroductionFrozen shoulder is a common, fibro-proliferative disease characterised by the insidious onset of pain and progressively restricted range of shoulder movement. Despite the prevalence of this disease, there is limited understanding of the molecular mechanisms underpinning the pathogenesis of this debilitating disease. Previous studies have identified increased myofibroblast differentiation and proliferation, immune cell influx and dysregulated cytokine production. We hypothesised that subpopulations within the stromal compartment may take on an activated phenotype, thus initiating the inflammatory processes observed in frozen shoulder. Therefore, we sought to evaluate the presence and possible pathogenic role of known stromal activation proteins in Frozen shoulder,MethodsShoulder capsule samples were collected from 10 patients with idiopathic frozen shoulder and 10 patients undergoing shoulder stabilisation surgery. Stromal activation marker expression (CD248, CD146, VCAM and PDPN, FAP) was quantified using immunohistochemistry. Control and diseased fibroblasts were cultured for in vitro studies from capsule biopsies from instability and frozen shoulder surgeries, respectively. The inflammatory profile and effects of IL-1β upon diseased and control fibroblasts was assessed using ELISA, immunohistochemistry and qPCR.ResultsImmunohistochemistry demonstrated increased expression of stromal activation markers CD248, CD146, VCAM and PDPN in the frozen shoulder group compared with control (p < 0.05). Fibroblasts cultured from diseased capsule produced elevated levels of inflammatory protein (IL-6, IL-8 & CCL-20) in comparison to control fibroblasts. Exposing control fibroblasts to an inflammatory stimuli, (IL-1ß) significantly increased stromal activation marker transcript and protein expression (CD248, PDPN and VCAM).ConclusionsThese results show that stromal fibroblasts have an activated phenotype in frozen shoulder and this is associated with inflammatory cytokine dysregulation. Furthermore, it supports the hypothesis that activated stromal fibroblasts may be involved in regulating the inflammatory and fibrotic processes involved in this disease.

scholarly journals P03.25 Neutralizing extracellular CHP-1 impairs tumor growth and metastasis formation

2020 ◽  
Vol 8 (Suppl 2) ◽  
pp. A32.2-A33
Author(s):  
L Seclì ◽  
F Fusella ◽  
M Brancaccio
Keyword(s):  
Cancer Progression ◽  
Immune Modulation ◽  
Molecular Mechanisms ◽  
Immune Cell ◽  
Tumor Grade ◽  
Metastasis Formation ◽  
Cancer Subtypes ◽  
Cancer Hallmarks ◽  
Clinical Models

BackgroundFound in the extracellular compartment, Heat Shock Proteins (HSPs) are actively secreted proteins that modulate the tumor behavior. Extracellular HSPs play a unique role as extracellular chaperones and receptors-binding molecules, favoring the establishment and maintenance of different cancer hallmarks, including immune modulation and evasion. CHP-1, is a ubiquitously expressed protein with chaperone activity and its high expression correlates with high tumor grade and lymph node positivity in different breast and lung cancer subtypes. In addition, CHP-1 is actively and uncanonically secreted by cancer cells in the tumor microenvironment (TME).Materials and MethodsSera cancer patients were analyzed for the presence of CHP-1. To assess the role of extracellular CHP-1 (eCHP-1) in the TME, in vitro experiments on different cell populations have been performed. To dissect the molecular mechanisms, through which eCHP-1 induces cancer progression, have been analyzed specific signaling pathways in cancer and immune cells. Immune cell composition in presence of eCHP-1 in tumors has been identified using flow-cytometry. The characterization of eCHP-1 inhibition as therapeutic approach has been conducted in breast and colon cancer pre-clinical models.ResultseCHP-1 activates an autocrine signaling through TLR2, TLR4 and LRP1, promoting tumor progression and metastasis formation in different pre-clinical models. Moreover, eCHP-1 can modulate the immune composition of the TME, making interesting the analysis of its inhibition in cancer immunotherapy.ConclusionseCHP-1 represents a easy accessible protein for diagnosis and targeting in very aggressive canncers.Disclosure InformationL. Seclì: None. F. Fusella: None. M. Brancaccio: None.

5952The involvement and interplay of HMGB1 with soluble MD-2 in dilated cardiomyopathy and its impact in immune cell recruitment

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
A Kuemmel ◽  
R Feldtmann ◽  
A Stohbach ◽  
A Riad ◽  
B Chamling ◽  
...  
Keyword(s):  
Hospital Admission ◽  
Dilated Cardiomyopathy ◽  
Molecular Mechanisms ◽  
Mononuclear Cells ◽  
Immune Cell ◽  
Early Death ◽  
Monocyte Adhesion ◽  
The Impact ◽  
Death Group

Abstract Objective Dilated cardiomyopathy (DCM) is characterized by systolic dysfunction and simultaneous dilatation of the left or both ventricles. Besides other causes, the innate immune system plays a major role in the development and progression of the disease. To uncover links between molecular mechanisms and disease progression our group has focused on the toll like receptor 4 / myeloid differentiation factor-2 (MD-2) system. Purpose We already reported that soluble MD-2 (sMD-2) is a risk factor for survival in patients with DCM. High mobility group box protein 1 (HMGB1) is a potent intrinsic interaction partner of MD-2. In the current study, we quantified HMGB-1 in plasma from patients with DCM at baseline, upon first hospital admission. Furthermore, we studied the impact of different HMGB-1 isoforms on monocyte adhesion in vitro. Methods We included 77 DCM patients divided by median time point of death after first hospital admission into “early death”, “late death” and “alive” group. MD-2 was quantified by means of ELISA. MD-2 and HMGB1 was quantified by means of ELISA. Statistical analysis was performed using a linear regression model. Human umbilical vein endothelial cells (HUVEC; n=6) were treated for 48h with two isoforms of HMGB1 (disulfide (ds) and fully reduced (fr)) alone and in combination with MD-2. Subsequently, those activated HUVEC were incubated with fresh isolated peripheral blood mononuclear cells (PBMCs) for 20 min. Finally, monocyte adhesion was quantified using multicolour FACS. Results At baseline, we found significantly increased sMD-2 level in the “early death” group (591.3±75.5 ng/ml) compared to the “later death” group (369.2±46.5 ng/ml; p=0.015) and the “alive” group (303.2±18.1 ng/ml; p<0.001). Likewise, we could demonstrate significantly increased levels of HMGB1 in the “early death” group (0.93±0.14 ng/ml) compared to the “later death” (0.57±0.17 ng/ml; p=0.04) and the “alive” group (0.49±0.06 ng/ml; p<0.001). In all patients who died during the observation period, sMD-2 and HMGB1 plasma levels showed a positive correlation. In vitro, we could demonstrate a significantly increased monocyte adhesion on HUVECs in the dsHMGB1 and the frHMGB1 group compared to controls (p=0.001; p=0.004). In contrast, the dsHMGB1 MD-2 group showed a significantly decreased monocyte adhesion on HUVECs compared to dsHMGB1 treatment alone (p=0.049). In the frHMGB1 MD-2 group, however, the reduction of the monocyte adhesion was less pronounced and did not reach significance (Fig. 1). Conclusion Our findings give a first hint that the interplay between HMGB1 and MD-2 is particularly involved in the development and progression of DCM. Furthermore, the data suggest that soluble MD-2 is capable of reducing the pro-inflammatory effects of dsHMGB1 but not of frHMGB1

scholarly journals Human immunodeficiency virus infection of human bone marrow stromal fibroblasts

Blood ◽  
1990 ◽  
Vol 76 (2) ◽  
pp. 317-322 ◽  
Author(s):  
DT Scadden ◽  
M Zeira ◽  
A Woon ◽  
Z Wang ◽  
L Schieve ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Human Immunodeficiency Virus ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Stromal Fibroblast ◽  
Stromal Fibroblasts ◽  
Fibroblast Line ◽  
Immunodeficiency Virus

Abstract The human immunodeficiency virus (HIV) preferentially infects CD4 positive T cells and monocytes. Other human cell types have been reported to be infectable with HIV, including cells of mesenchymal origin. In this report, we show that both primary human bone marrow stromal fibroblasts and an immortalized human stromal fibroblast line are susceptible to HIV infection. These cells are capable of passing HIV to cells of lymphoid or myeloid lineage, and may thereby act as a reservoir of virus. This in vitro system may be a useful model for assessing the pathophysiology of hematopoietic dysfunction in AIDS patients.

scholarly journals Anti-Inflammatory and Immunosuppressive Effects of the A2AAdenosine Receptor

2011 ◽  
Vol 11 ◽  
pp. 320-339 ◽  
Author(s):  
Gillian R. Milne ◽  
Timothy M. Palmer
Keyword(s):  
Molecular Mechanisms ◽  
Immune Cell ◽  
Inflammatory Responses ◽  
Current Evidence ◽  
Receptor Subtypes ◽  
Protective Effects ◽  
Stress Injury ◽  
Anti Inflammatory

The production of adenosine represents a critical endogenous mechanism for regulating immune and inflammatory responses during conditions of stress, injury, or infection. Adenosine exerts predominantly protective effects through activation of four 7-transmembrane receptor subtypes termed A1, A2A, A2B, and A3, of which the A2Aadenosine receptor (A2AAR) is recognised as a major mediator of anti-inflammatory responses. The A2AAR is widely expressed on cells of the immune system and numerousin vitrostudies have identified its role in suppressing key stages of the inflammatory process, including leukocyte recruitment, phagocytosis, cytokine production, and immune cell proliferation. The majority of actions produced by A2AAR activation appear to be mediated by cAMP, but downstream events have not yet been well characterised. In this article, we review the current evidence for the anti-inflammatory effects of the A2AAR in different cell types and discuss possible molecular mechanisms mediating these effects, including the potential for generalised suppression of inflammatory gene expression through inhibition of the NF-κB and JAK/STAT proinflammatory signalling pathways. We also evaluate findings fromin vivostudies investigating the role of the A2AAR in different tissues in animal models of inflammatory disease and briefly discuss the potential for development of selective A2AAR agonists for use in the clinic to treat specific inflammatory conditions.

scholarly journals Network models of primary melanoma microenvironments identify key melanoma regulators underlying prognosis

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Won-Min Song ◽  
Praveen Agrawal ◽  
Richard Von Itter ◽  
Barbara Fontanals-Cirera ◽  
Minghui Wang ◽  
...  
Keyword(s):  
Large Scale ◽  
Molecular Mechanisms ◽  
Immune Cell ◽  
Primary Melanoma ◽  
Network Models ◽  
Transcriptional Networks ◽  
Melanoma Progression ◽  
Skin Malignancy

AbstractMelanoma is the most lethal skin malignancy, driven by genetic and epigenetic alterations in the complex tumour microenvironment. While large-scale molecular profiling of melanoma has identified molecular signatures associated with melanoma progression, comprehensive systems-level modeling remains elusive. This study builds up predictive gene network models of molecular alterations in primary melanoma by integrating large-scale bulk-based multi-omic and single-cell transcriptomic data. Incorporating clinical, epigenetic, and proteomic data into these networks reveals key subnetworks, cell types, and regulators underlying melanoma progression. Tumors with high immune infiltrates are found to be associated with good prognosis, presumably due to induced CD8+ T-cell cytotoxicity, via MYO1F-mediated M1-polarization of macrophages. Seventeen key drivers of the gene subnetworks associated with poor prognosis, including the transcription factor ZNF180, are tested for their pro-tumorigenic effects in vitro. The anti-tumor effect of silencing ZNF180 is further validated using in vivo xenografts. Experimentally validated targets of ZNF180 are enriched in the ZNF180 centered network and the known pathways such as melanoma cell maintenance and immune cell infiltration. The transcriptional networks and their critical regulators provide insights into the molecular mechanisms of melanomagenesis and pave the way for developing therapeutic strategies for melanoma.

Albumin internalizes and inhibits endosomal TLR signaling in leukocytes from patients with decompensated cirrhosis

2020 ◽  
Vol 12 (566) ◽  
pp. eaax5135 ◽  
Author(s):  
Mireia Casulleras ◽  
Roger Flores-Costa ◽  
Marta Duran-Güell ◽  
José Alcaraz-Quiles ◽  
Silvia Sanz ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Immune Cell ◽  
Human Albumin ◽  
Decompensated Cirrhosis ◽  
Type I ◽  
Tlr Signaling ◽  
Cpg Dna ◽  
Cell Transcriptome

Human serum albumin (HSA) is an emerging treatment for preventing excessive systemic inflammation and organ failure(s) in patients with acutely decompensated (AD) cirrhosis. Here, we investigated the molecular mechanisms underlying the immunomodulatory properties of HSA. Administration of HSA to patients with AD cirrhosis with elevated circulating bacterial DNA rich in unmethylated cytosine-phosphate-guanine dideoxynucleotide motifs (CpG-DNA) was associated with reduced plasma cytokine concentrations. In isolated leukocytes, HSA abolished CpG-DNA–induced cytokine expression and release independently of its oncotic and scavenging properties. Similar anti-inflammatory effects were observed with recombinant human albumin. HSA exerted widespread changes on the immune cell transcriptome, specifically in genes related to cytokines and type I interferon responses. Our data revealed that HSA was taken up by leukocytes and internalized in vesicles positively stained with early endosome antigen 1 and colocalized with CpG-DNA in endosomes, where the latter binds to Toll-like receptor 9 (TLR9), its cognate receptor. Furthermore, HSA also inhibited polyinosinic:polycytidylic acid– and lipopolysaccharide-induced interferon regulatory factor 3 phosphorylation and TIR domain–containing adapter-inducing interferon-β–mediated responses, which are exclusive of endosomal TLR3 and TLR4 signaling, respectively. The immunomodulatory actions of HSA did not compromise leukocyte defensive mechanisms such as phagocytosis, efferocytosis, and intracellular reactive oxygen species production. The in vitro immunomodulatory effects of HSA were confirmed in vivo in analbuminemic humanized neonatal Fc receptor transgenic mice. These findings indicate that HSA internalizes in immune cells and modulates their responses through interaction with endosomal TLR signaling, thus providing a mechanism for the benefits of HSA infusions in patients with cirrhosis.

Elucidating Molecular Mechanisms of Cross-Presentation

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2217-2217
Author(s):  
Shin-Rong Julia Wu
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Immune System ◽  
Molecular Mechanisms ◽  
Conflicts Of Interest ◽  
Activation Markers ◽  
Mhc I ◽  
Cross Presentation ◽  
Intact Immune System

The immune system regulates many processes vital to homeostasis. Antigen cross-presentation, an immune process specific to dendritic cells (DCs), critically contributes to maintaining homeostasis by regulating immune tolerance, antiviral activity, and antitumor responses. Through cross-presentation, extracellular antigen is processed and presented by MHC I on DCs to CD8 T cells. Despite cross-presentation's crucial role in mediating immune responses, its molecular regulation remains poorly defined. A potential breakthrough came when, using an in vitro shRNA-mediated knockdown (KD) approach, a group identified Sec22b as a central regulatory molecule of the pathway1. They demonstrated that Sec22b mediates recruitment of proteins necessary for MHC I-antigen loading from the endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC) to the antigen-containing phagosome, promoting cross-presentation. Thus, we tested the hypothesis that Sec22b mediates cross-presentation in vivo, in the context of an intact immune system under physiological conditions. We generated DC-specific Sec22b knockout (KO) mice (CD11cCre Sec22bfl/fl) from Sec22b-conditional gene trapped founder mice. These mice develop normally and have an intact immune system. KO DCs from spleen and bone marrow (BM) have Cre-mediated excision at the Sec22b locus, verified by PCR, and reduction of Sec22b production, verified by Western Blot. KO DCs from spleen and BM express activation markers in response to TLR and NLR stimulation at comparable levels to Cre- Sec22bfl/fl (FL) mice. By adoptively transferring ovalbumin (OVA)-specific (OT-I) T cells into these mice, then injecting with soluble OVA i.p., we measured OT-I T cell proliferation as a readout of cross-presentation. To our surprise, we saw no difference in the ability of KO versus FL mice to cross-present OVA (p >0.9). This observation was verified with in vitro assays with KO DCs from BM and spleen cross-presenting soluble OVA (0-3 mg/mL). We obtained similar findings using bead-bound, insoluble OVA. From this, we concluded Sec22b is not necessary for cross-presentation, invalidating our hypothesis. We were, however, able to reduce cross-presentation by shRNA-mediated KD of Sec22b (p <0.05), reproducing published observations. This discrepancy in observations was not due to functional compensation in KO BMDCs by Sec22b homologs, Sec22a or Sec22c, which we determined using qPCR. Intriguingly, when we treated KO BMDCs with the Sec22b-targeting shRNA, we again observed a reduction in cross-presentation (p <0.05). The reduction was comparable to that found in Sec22b-targeting shRNA-treated FL and WT BMDCs. Taken together, our data (a) demonstrate that Sec22b is not necessary for cross-presentation, (b) suggest the existence of a novel critical mediator of cross-presentation that is also targeted by the shRNA sequence used and (c) caution against extrapolating mechanisms or phenotypes based on KD studies alone. Cebrian, I. et al. Sec22b regulates phagosomal maturation and antigen crosspresentation by dendritic cells. Cell147, 1355-1368 (2011). Disclosures No relevant conflicts of interest to declare.

scholarly journals IL-21 Signaling in Immunity

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 224 ◽  
Author(s):  
Warren J. Leonard ◽  
Chi-Keung Wan
Keyword(s):  
T Cells ◽  
Molecular Mechanisms ◽  
Immune Cell ◽  
Cell Types ◽  
Clinical Settings ◽  
Type I ◽  
Next Generation Sequencing Technology ◽  
Wide Range

IL-21 is a type I cytokine produced by T cells and natural killer T cells that has pleiotropic actions on a wide range of immune and non-immune cell types. Since its discovery in 2000, extensive studies on the biological actions of IL-21 have been performed in vitro and in vivo. Recent reports describing patients with primary immunodeficiency caused by mutations of IL21 or IL21R have further deepened our knowledge of the role of this cytokine in host defense. Elucidation of the molecular mechanisms that mediate IL-21’s actions has provided the rationale for targeting IL-21 and IL-21 downstream mediators for therapeutic purposes. The use of next-generation sequencing technology has provided further insights into the complexity of IL-21 signaling and has identified transcription factors and co-factors involved in mediating the actions of this cytokine. In this review, we discuss recent advances in the biology and signaling of IL-21 and how this knowledge can be potentially translated into clinical settings.

scholarly journals Altered Differentiation of Endometrial Mesenchymal Stromal Fibroblasts Is Associated With Endometriosis Susceptibility

2021 ◽  
Author(s):  
Brett McKinnon ◽  
Samuel Lukowski ◽  
Sally Mortlock ◽  
Joanna Crawford ◽  
Rebecca Johnston ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cellular Differentiation ◽  
Disease Susceptibility ◽  
Mesenchymal Cell ◽  
Stromal Fibroblast ◽  
Stromal Fibroblasts ◽  
Endometrial Cells ◽  
Growth Profiles

Abstract Cellular development is tightly regulated as mature cells with aberrant functions may initiate pathogenic processes. The endometrium is a highly regenerative tissue, shedding and regenerating each month. Endometrial stromal fibroblasts are regenerated each cycle from mesenchymal stem cells and play a pivotal role in endometriosis, a disease characterised by endometrial cells that grow outside the uterus. Why the cells of some women are more capable of developing into endometriosis lesions is not clear. Using isolated, purified and cultured endometrial cells of mesenchymal origin from 19 women with (n = 10) and without (n = 9) endometriosis we analysed the transcriptome of 33,758 individual cells and compared these to clinical characteristics and in vitro growth profiles. We show purified mesenchymal cell cultures include a mix of mesenchymal stem cells and two endometrial stromal fibroblast subtypes with distinct transcriptomic signatures indicative of varied progression through the differentiation processes. The fibroblast subgroup characterised by incomplete differentiation was predominantly (81%) derived from women with endometriosis and exhibited an altered in vitro growth profile. These results uncover an inherent difference in endometrial cells of women with endometriosis and highlight the relevance of cellular differentiation and its potential to contribute to disease susceptibility.

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