The IL1-like cytokine IL33 and its receptor ST2 are abnormally expressed in the affected skin and visceral organs of patients with systemic sclerosis

2009 ◽  
Vol 69 (3) ◽  
pp. 598-605 ◽  
Author(s):  
Mirko Manetti ◽  
Lidia Ibba-Manneschi ◽  
Vasiliki Liakouli ◽  
Serena Guiducci ◽  
Anna Franca Milia ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Rt Pcr ◽  
Interleukin 33 ◽  
Pulmonary Fibroblasts ◽  
Confocal Immunofluorescence Microscopy ◽  
Confocal Immunofluorescence ◽  
Skin Biopsies ◽  
Activated Fibroblasts ◽  
St2 Receptor

BackgroundEarly endothelial cell (EC) activation/damage and profibrotic Th2-associated cytokines play a pivotal role in systemic sclerosis (SSc). Interleukin 33 (IL33) is a novel member of the IL1 family that promotes Th2 responses and inflammation through the ST2 receptor. IL33 is also a chromatin-associated transcriptional regulator in ECs.ObjectiveTo investigate the role of the IL33/ST2 axis in SSc.MethodsSkin biopsies were obtained from 30 patients with SSc (15 early/15 late stage) and 10 healthy subjects. Lung, kidney, heart, oesophagus, stomach, placenta biopsies and bronchoalveolar lavage cells from patients with SSc and controls were also analysed. IL33/ST2 expression was investigated by immunohistology, confocal immunofluorescence microscopy, western blotting and RT-PCR.ResultsIn skin biopsies from control subjects, constitutive nuclear IL33 protein expression was found in dermal ECs and keratinocytes, while ST2 was weakly expressed in ECs and fibroblasts. In skin biopsies from patients with early SSc, IL33 protein was downregulated or absent in ECs and epidermis while IL33 mRNA was normally expressed or even upregulated. Moreover, ECs, perivascular infiltrating mast cells, CD68-positive macrophages, CD3-positive T cells, CD20-positive B cells and activated fibroblasts/myofibroblasts exhibited strong ST2 expression. In skin biopsies from patients with late SSc, IL33 was constitutively found in most ECs while ST2 immunostaining was weaker. In early SSc, the loss of endothelial IL33 protein and the overexpression of ST2 involved all affected organs. Dermal and pulmonary fibroblasts showed IL33 expression in SSc.ConclusionIL33 and ST2 are abnormally expressed in SSc. In early SSc, upon EC activation/damage IL33 may be mobilised from ECs to signal through ST2 in key profibrotic players such as inflammatory/immune cells and fibroblasts/myofibroblasts.

HLA-G Expression in the Skin of Patients with Systemic Sclerosis

2009 ◽  
Vol 36 (6) ◽  
pp. 1230-1234 ◽  
Author(s):  
ISABELA J. WASTOWSKI ◽  
PERCIVAL D. SAMPAIO-BARROS ◽  
ELIANE M.I. AMSTALDEN ◽  
GUSTAVO MARTELLI PALOMINO ◽  
JOÃO FRANCISCO MARQUES-NETO ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Control Sample ◽  
Healthy Controls ◽  
Skin Disorders ◽  
Disease Prognosis ◽  
Laboratory Variables ◽  
Skin Biopsies ◽  
Dermal Cells

Objective.To determine HLA-G expression in skin biopsies from patients with systemic sclerosis (SSc), and its association with epidemiological, clinical, and laboratory variables and survival.Methods.Paraffin-embedded skin biopsies obtained from 21 SSc patients (14 limited SSc, 7 diffuse SSc) and from 28 healthy controls were studied. HLA-G expression was evaluated by immunohistochemistry.Results.HLA-G molecules were detected in 57% of skin biopsies from patients with SSc (9 from limited SSc, 3 from diffuse SSc), whereas no control sample expressed HLA-G (p = 0.000004). In patients, HLA-G molecules were consistently observed within epidermal and some dermal cells. HLA-G expression was associated with a lower frequency of vascular cutaneous ulcers (p = 0.0004), telangiectasias (p = 0.008), and inflammatory polyarthralgia (p = 0.02). After a 15-year followup, SSc patients who exhibited HLA-G survived longer than patients who did not.Conclusion.HLA-G is expressed in skin biopsies from patients with SSc, and this is associated with a better disease prognosis. This suggests a modulatory role of HLA-G in SSc, as observed in other skin disorders.

Abstract 16191: Phospholamban Regulates Perinuclear/nuclear Calcium Dynamics in Cardiomyocytes

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Adonis Z Wu ◽  
Yi-Hsin Chan ◽  
Shien-Fong Lin ◽  
Peng-Sheng Chen ◽  
Zhenhui Chen
Keyword(s):  
Immunofluorescence Microscopy ◽  
Fab Fragment ◽  
Cardiac Sarcoplasmic Reticulum ◽  
Confocal Immunofluorescence Microscopy ◽  
Slow Decay ◽  
Confocal Immunofluorescence ◽  
Intracellular Ca ◽  
Nuclear Regions ◽  
Uptake And Release

Introduction: Phospholamban (PLB) regulates cardiac sarcoplasmic reticulum (SR) Ca 2+ -ATPase (SERCA2a), thus modulating SR Ca 2+ dynamics. Recent studies demonstrated that SERCA is involved in Ca 2+ uptake into the lumen of nuclear envelope (NE) of cardiomyocytes (CMs). However, the regulatory role of PLB on Ca 2+ uptake into NE remains unknown. Hypothesis: We hypothesize that PLB is also responsible for modulating nuclear Ca 2+ dynamics. Methods: Confocal immunofluorescence microscopy was used to determine subcellular expression of PLB. By using fluo-4 based confocal line-scan Ca 2+ imaging, we measured spontaneous Ca 2+ waves (SCWs) across both cytoplasmic and nuclear regions in isolated permeabilized mouse CMs. Results: Several anti-PLB antibodies strongly stained PLB at both SR and the perinuclear membranes in CMs. A PLB peptide (residues 1-31) eliminated all these anti-PLB antibody stains. To identify the functional role of PLB expressed in the perinuclear membranes, we took advantage of our recently established method that a Fab fragment of anti-PLB monoclonal antibody (Fab) reversed PLB inhibition specifically and increased SR Ca 2+ uptake and release. SCWs through the nuclear regions had typically relative low fluorescent amplitude (F/F 0 ) and slow decay time (t 1/2 ) compared to that in the cytoplasmic region. At the free intracellular Ca 2+ concentration ([Ca 2+ ] i ) of 400 nM, Fab (100 μg/mL) significantly enhanced F/F 0 and decreased t 1/2 of SCWs in both cytoplasmic and nuclear regions. After addition of Fab, F/F 0 of SCWs through the nuclear regions increased from 0.91±0.16 to 1.27±0.19 (n=9, p<.05) while t 1/2 decreased from 137.6±18.5 ms to 105.0±11.3 ms, (p<.05). Similar effects were also observed after phosphorylation of PLB by addition of 20 μM cAMP (F/F 0 =1.43±0.11 vs. 1.04±0.14 in control, p<.05; t 1/2 =107.82±10.9 ms vs. 139.21±20.1 ms in control, n=6, p<.05). At high [Ca 2+ ] i of 1000 nM where PLB does not inhibit SERCA2a, addition of cAMP or Fab had no significant effect on SCWs in both cytoplasmic and nuclear regions. Conclusions: We demonstrated that PLB is expressed in and around NE. Acute removal of PLB inhibition increased perinuclear/nuclear Ca 2+ uptake and release. PLB is critically involved in nuclear Ca 2+ signaling modulation.

scholarly journals HHV-6A Infection and Systemic Sclerosis: Clues of a Possible Association

2019 ◽  
Vol 8 (1) ◽  
pp. 39
Author(s):  
Elisabetta Caselli ◽  
Irene Soffritti ◽  
Maria D’Accolti ◽  
Daria Bortolotti ◽  
Roberta Rizzo ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Plasma Levels ◽  
Human Herpesvirus ◽  
Infectious Agents ◽  
Internal Organs ◽  
Matrix Deposition ◽  
Skin Biopsies ◽  
Extracellular Matrix Deposition

Systemic sclerosis (SSc) is an autoimmune disease characterized by vasculopathy, excessive extracellular matrix deposition, and fibrosis of the skin and internal organs. Several infectious agents, including human herpesvirus-6 (HHV-6), have been suggested as possible triggering factors, but a direct association is still missing. We characterized 26 SSc patients for the presence of HHV-6 in tissues and blood, the anti-HHV-6 response, HLA-G plasma levels, and KIR typing. Given the prominent role of endothelial cells (EC) in SSc pathogenesis, along with HHV-6 tropism for EC, we also investigated the expression of pro-fibrosis factors in HHV-6 infected EC. Results showed the presence of HHV-6A in skin biopsies, and an increased virus load was associated with disease severity and poor natural killer (NK) response against the virus, particularly in subjects exhibiting a KIR2 phenotype. HLA-G plasma levels were significantly higher in HHV-6A/B-KIR2 positive SSc patients and in vitro HHV-6A infection-induced pro-fibrosis factors expression in EC, supporting its role in the development of the fibrosing process. Our data suggest an association between virus infection/reactivation and disease, opening the way to future studies to understand the mechanisms by which HHV-6A might contribute to the multifactorial pathogenesis of SSc.

scholarly journals Expression of the adaptor protein m-Numb in mouse male germ cells

Reproduction ◽  
2006 ◽  
Vol 132 (6) ◽  
pp. 887-897 ◽  
Author(s):  
Serena Corallini ◽  
Stefania Fera ◽  
Laura Grisanti ◽  
Ilaria Falciatori ◽  
Barbara Muciaccia ◽  
...  
Keyword(s):  
Cell Fate ◽  
Germ Cells ◽  
Adaptor Protein ◽  
Notch Signaling Pathway ◽  
Seminiferous Tubules ◽  
Confocal Immunofluorescence Microscopy ◽  
Confocal Immunofluorescence ◽  
Evolutionarily Conserved ◽  
Asymmetric Partitioning

Numb is an adaptor protein that is asymmetrically inherited at mitosis and controls the fate of sibling cells in different species. The role of m-Numb (mammalian Numb) as an important cell fate-determining factor has extensively been described mostly in neural tissues, particularly in progenitor cells, in the mouse. Biochemical and genetic analyses have shown that Numb acts as an inhibitor of the Notch signaling pathway, an evolutionarily conserved pathway involved in the control of cell proliferation, differentiation, and apoptosis. In the present study, we sought to determine m-Numb distribution in germ cells in the postnatal mouse testis. We show that all four m-Numb isoforms are widely expressed during postnatal testis development. By reverse transcriptase-PCR and western blot analyses, we further identify p71 as the predominantly expressed isoform in germ cells. Moreover, we demonstrate through co-immunoprecipitation studies that m-Numb physically associates with Ap2a1, a component of the endocytotic clathrin-coated vesicles. Finally, we employed confocal immunofluorescence microscopy of whole mount seminiferous tubules and isolated germ cells to gain more insight into the subcellular localization of m-Numb. These morphological analyses confirmed m-Numb and Ap2a1 co-localization. However, we did not observe asymmetric localization of m-Numb neither in mitotic spermatogonial stem cells nor in more differentiated spermatogonial cells, suggesting that spermatogonial stem cell fate in the mouse does not rely on asymmetric partitioning of m-Numb.

What is the role of interleukin 33 and ST2 receptor in myasthenia gravis?

2018 ◽  
Vol 315 ◽  
pp. 50-57 ◽  
Author(s):  
Izabela Monika Rozmilowska ◽  
Monika Helena Adamczyk-Sowa
Keyword(s):  
Myasthenia Gravis ◽  
Interleukin 33 ◽  
St2 Receptor

scholarly journals EMILIN proteins are novel extracellular constituents of the dentin-pulp complex

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Thomas Imhof ◽  
Yüksel Korkmaz ◽  
Manuel Koch ◽  
Gerhard Sengle ◽  
Alvise Schiavinato
Keyword(s):  
Developmental Stages ◽  
Confocal Immunofluorescence Microscopy ◽  
Confocal Immunofluorescence ◽  
Associated Proteins ◽  
Supramolecular Networks ◽  
First Time ◽  
Early Developmental ◽  
Caries Lesions ◽  
Pulp Biology

Abstract Odontoblasts and pulp stroma cells are embedded within supramolecular networks of extracellular matrix (ECM). Fibrillin microfibrils and associated proteins are crucial constituents of these networks, serving as contextual scaffolds to regulate tissue development and homeostasis by providing both structural and mechanical properties and sequestering growth factors of the TGF-β superfamily. EMILIN-1, -2, and -3 are microfibril-associated glycoproteins known to modulate cell behaviour, growth factor activity, and ECM assembly. So far their expression in the various cells of the dentin-pulp complex during development, in the adult stage, and during inflammation has not been investigated. Confocal immunofluorescence microscopy and western blot analysis of developing and adult mouse molars and incisors revealed an abundant presence of EMILINs in the entire dental papilla, at early developmental stages. Later in development the signal intensity for EMILIN-3 decreases, while EMILIN-1 and -2 staining appears to increase in the pre-dentin and in the ECM surrounding odontoblasts. Our data also demonstrate new specific interactions of EMILINs with fibulins in the dentin enamel junction. Interestingly, in dentin caries lesions the signal for EMILIN-3 was significantly increased in inflamed odontoblasts. Overall our findings point for the first time to a role of EMILINs in dentinogenesis, pulp biology, and inflammation.

SAT0294 IL33 ACTIVATES FIBROBLASTS AND INDUCES SKIN FIBROSIS IN SYSTEMIC SCLEROSIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1091.1-1092
Author(s):  
X. Wu ◽  
B. Ming ◽  
L. Dong
Keyword(s):  
Systemic Sclerosis ◽  
Dermal Fibroblast ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Skin Fibrosis ◽  
Human Dermal Fibroblasts ◽  
Interleukin 33 ◽  
Dermal Fibrosis

Background:Systemic sclerosis (SSc) is a chronic immune-mediated autoimmune disease that is characterized by fibrotic changes of the skin and internal organs, which in turn leads to distortion of tissue structure and gradual loss of organ function. So far, there is still no treatment allows full recovery from this severe disorder. Therefore, it is of great social significance to study the pathogenesis of this disease and find new targets for treatment. Interleukin 33 (IL-33), which is a potent inducer of type 2 immune response, has been confirmed to be involved in the development and progression of multiple fibrotic diseases. However, the role and mechanism of IL-33 in SSc-related fibrosis remains unclear.Objectives:To clarify the role of interleukin 33 (IL-33) and its receptor Suppression of tumorigenicity 2 (ST2) in the skin fibrosis of SSc, so to provides a new target for the treatment of fibrosis in patients with SSc.Methods:The levels of IL-33 and ST2 was analysed in human samples, murine models of SSc and in cultured fibroblasts by immunohistochemistry and immunofluorescence. The functional role of IL-33 was evaluated by detecting changes in proliferation, migration, and activation of fibroblasts stimulated with recombinant IL-33 protein. MAPK and NF-κB signallings of fibroblasts were assessed by western blotting and analyses of target genes. The role of IL-33 in skin fibrosis was analysed in IL-33 deficient mice (il33−/−) and wild-type controls injected with bleomycin or NaCl.Results:The expression of IL-33 and its receptor ST2 were up-regulated in skin lesions of SSc patients (Fig 1 A-C) and bleomycin-treated mice(Fig1 D-F). Compared to the healthy skin, the skin from SSc patients expressed more ST2 on fibroblasts membrane(Fig 1 B-C). IL33 induces MAPK and IκBα activation in human dermal fibroblast(Fig 2 A), and promote proliferation, migration and production of collagen of human dermal fibroblasts, but not the release of inflammatory factors(IL-6, MCP-1)(Fig2 B-G). Mice deficient for IL33 are protected from bleomycin-induced dermal fibrosis (Fig3).Fig 1.Increased expression of IL33, ST2 in SSc patients and bleomycin-treated mice.Fig 2.IL33 induces MAPK and IκBα activation in human dermal fibroblast, and and promote proliferation, migration and production of collagen of human dermal fibroblasts.Fig 3.Mice deficient for IL33 are protected from bleomycin-induced dermal fibrosis.Conclusion:IL33 promotes skin fibrosis by activating fibroblasts, and IL33/ST2 may be an important target for the treatment of fibrosis in patients with SSc.References:[1]Ingegnoli F, Ughi N, Mihai C. Update on the epidemiology, risk factors, and disease outcomes of systemic sclerosis. Best practice & research. Clinical rheumatology. 2018;32(2):223-240.[2]Schmitz J, Owyang A, Oldham E, et al. IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines. Immunity. 2005;23(5):479-490.[3]Molofsky AB, Savage AK, Locksley RM. Interleukin-33 in Tissue Homeostasis, Injury, and Inflammation.Immunity.2015;42(6):1005-1019.Disclosure of Interests:None declared

The role of pulmonary fibroblasts in alveolar septation

Pneumologie ◽  
2012 ◽  
Vol 66 (S 01) ◽  
Author(s):  
F Klein ◽  
A Wilde ◽  
W Seeger ◽  
R Voswinckel
Keyword(s):  
Pulmonary Fibroblasts

Monitoring the marine environment for small round structured viruses (SRSVS): a new approach to combating the transmission of these viruses by molluscan shellfish

1998 ◽  
Vol 38 (12) ◽  
pp. 51-56 ◽  
Author(s):  
K. Henshilwood ◽  
J. Green ◽  
D. N. Lees
Keyword(s):  
Enteric Virus ◽  
Winter Period ◽  
Rt Pcr ◽  
Cloning And Sequencing ◽  
New Approach ◽  
Human Enteric Virus ◽  
Different Strains ◽  
The Uk ◽  
Public Health Protection

This study investigates human enteric virus contamination of a shellfish harvesting area. Samples were analysed over a 14-month period for Small Round Structured Viruses (SRSVs) using a previously developed nested RT-PCR. A clear seasonal difference was observed with the largest numbers of positive samples obtained during the winter period (October to March). This data concurs with the known winter association of gastroenteric illness due to oyster consumption in the UK and also with the majority of the outbreaks associated with shellfish harvested from this area during the study period. RT-PCR positive amplicons were further characterised by cloning and sequencing. Sequence analysis of the positive samples identified eleven SRSV strains, of both Genogroup I and Genogroup II, occurring throughout the study period. Many shellfish samples contained a mixture of strains with a few samples containing up to three different strains with both Genogroups represented. The observed common occurrence of strain mixtures may have implications for the role of shellfish as a vector for dissemination of SRSV strains. These results show that nested RT-PCR can identify SRSV contamination in shellfish harvesting areas. Virus monitoring of shellfish harvesting areas by specialist laboratories using RT-PCR is a possible approach to combating the transmission of SRSVs by molluscan shellfish and could potentially offer significantly enhanced levels of public health protection.

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