scholarly journals Activation of the Interferon Pathway in Peripheral Blood of Patients with Sjögren’s Syndrome

2010 ◽  
Vol 38 (2) ◽  
pp. 310-316 ◽  
Author(s):  
OSAMU KIMOTO ◽  
JIN SAWADA ◽  
KUMIKO SHIMOYAMA ◽  
DAISUKE SUZUKI ◽  
SATOKI NAKAMURA ◽  
...  
Keyword(s):  
Real Time ◽  
Microarray Analysis ◽  
Dna Microarray ◽  
Real Time Pcr ◽  
Peripheral Blood ◽  
Sjogren’S Syndrome ◽  
Sjogren's Syndrome ◽  
Expression Level ◽  
Healthy Controls ◽  
Dna Microarray Analysis

Objective.DNA microarray analysis and quantitative real-time polymerase chain reaction (PCR) were performed to identify key target genes in peripheral blood from patients with Sjögren’s syndrome (SS).Methods.DNA microarray analysis was performed in 19 patients with SS (all women) and 10 healthy controls (5 men and 5 women) using a low-density DNA microarray system with 778 genes. For confirmation, the expression of upregulated genes was analyzed by quantitative real-time PCR in another 37 SS patients (35 women and 2 men) and 9 healthy controls (8 women and 1 man). Relationships between gene signatures and various clinical measures, such as disease duration, symptoms and signs, complications, immunological findings, and salivary and lacrimal functions, were analyzed.Results.Interferon-α (IFN-α)-inducible protein 27 (IFI27) showed the most significant difference between SS patients and controls in the microarray screening. We performed quantitative RT-PCR for IFI27. IFI27 gene expression level was increased in patients with SS compared with controls (p < 0.01) by real-time PCR, supporting our observations from the microarray data. The level of IFI27 was significantly correlated with serum IgG levels (r = 0.462, p < 0.01) and ß2-microglobulin (r = 0.385, p < 0.05), soluble interleukin 2 receptor (r = 0.473, p < 0.01), erythrocyte sedimentation rate (r = 0.333, p < 0.05), and antinuclear antibody titer (speckled pattern; r = 0.445, p < 0.01).Conclusion.Our results suggest that upregulation of IFN-inducible genes in SS patients is a systemic phenomenon, and IFN may play an important role in the pathogenesis of SS. The expression level of IFI27 could be an effective and specific biomarker associated with SS.

Levels of plasmacytoid dendritic cells and type-2 myeloid dendritic cells are reduced in peripheral blood of patients with primary Sjögren's syndrome

2009 ◽  
Vol 69 (6) ◽  
pp. 1235-1238 ◽  
Author(s):  
Petra Vogelsang ◽  
Johan G Brun ◽  
Gunnvor Øijordsbakken ◽  
Kathrine Skarstein ◽  
Roland Jonsson ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Sjögren’S Syndrome ◽  
Peripheral Blood ◽  
Sjögren's Syndrome ◽  
Sjogren’S Syndrome ◽  
Sjogren's Syndrome ◽  
Healthy Controls ◽  
Myeloid Dendritic Cells ◽  
Antigen Presenting ◽  
Sjögren‘S Syndrome

ObjectiveSjögren's syndrome (SS) is a lymphoproliferative autoimmune disease, characterised by dryness of the mouth and eyes. Dendritic cells (DC) are potent antigen-presenting cells crucial for initiating and maintaining primary immune responses. This study quantified interferon-producing plasmacytoid DC (pDC) and two myeloid DC subsets (mDC1 and mDC2) in peripheral blood (PB) from primary SS (pSS) patients and healthy controls.MethodsBlood samples from 31 pSS patients and 28 gender and age-matched healthy controls were analysed by flow cytometry using the Miltenyi Blood DC enumeration kit. The presence of pDC in salivary glands (SG) from pSS patients was analysed by immunohistochemistry.ResultsPatients with pSS had significantly less pDC and mDC2 in PB compared with healthy controls. Moreover, pDC are present in SG from patients with pSS.ConclusionPatients with pSS have alterations among DC populations in PB, and pDC are present in the SG, suggesting a potential role of these cells in SS.

A Comparative Gene Expression Study of CD3+ T-Cells In Bone Marrow From Patients with Chronic ITP and Healthy Controls by DNA Microarray.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1445-1445
Author(s):  
Margareta Jernås ◽  
Bob Olsson ◽  
Hans Wadenvik
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Cell Adhesion ◽  
T Cell ◽  
Microarray Analysis ◽  
Dna Microarray ◽  
Adhesion Molecule ◽  
Healthy Controls ◽  
Dna Microarray Analysis ◽  
Chronic Itp

Abstract Abstract 1445 Background: The pathophysiology of ITP (immune thrombocytopenia) is more complex than initially believed. The importance of regulatory T-cells, platelet specific cytotoxic T-cells, suppression of megakaryocyte proliferation and maturation, functional thrombopoietin deficiency and inappropriately low bone marrow production of platelets has been unraveled over the last decade. T-cells are important in all forms of autoimmunity including ITP. However, very little is known about the T-cell activity in organs where platelets and their precursors are targeted by effectors of the immune system. To provide new insights into the pathophysiology of ITP, we performed a DNA microarray analysis on isolated bone marrow derived T-cells from chronic ITP patients and healthy controls. Patients and Method: Bone marrow was collected from 6 chronic ITP-patients (3 males and 3 females, mean age 39.5 years) and 6 healthy controls (3 males and3 females, mean age 33.8 years) through iliac crest bone marrow aspiration. T-cells were isolated by immunomagnetic cell sorting (Miltenyi Biotec, Surrey, UK). RNA was prepared using the Chomczynski method (Chomczynski and Sacchi, 1987), followed by RNeasy minielute clean-up (Qiagen, Hilden, Germany). RNA was amplified from 20 ng of starting total RNA with the Ovation RNA Amplification System V2 (NuGEN Technologies, Inc.), and cDNA was synthesized using the Encore Biotin Module kit (NuGEN Technologies, Inc.). After standard labeling, each sample was hybridized to an Affymetrix U133Plus 2.0 Human Genome array (Santa Clara, CA). Results: We identified 397 differently regulated genes in the bone marrow derived T-cells between ITP patients and controls by the DNA microarray analysis (P-value <0.05 and fold change ± 1.3). Some of the regulated genes have previously been implicated in ITP, such as integrin alpha 4 (ITGA4 or CD49D or VLA-4), Fas ligand (FASLG) and cytotoxic lymphocyte-associated 4 (CTLA-4). Furthermore, several apoptotic genes were lower expressed in ITP patients, such as Fas apoptotic inhibitory molecule 2 (FAIM2), BCL2-like 11 (BCL2L11), BCL2-like 13 (BCL2L13), peptidyl-tRNA hydrolase 2 (PTRH2) and the enzyme inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta (IKBKB). Integrin alpha 6, (ITGA6) and the adhesion molecule endothelial cell adhesion molecule (ESAM) were higher expressed in ITP patients compared to controls and are important for T-cell adhesion and migration or “homing” of lymphocytes to different lymphoid organs and inflammatory sites. Conclusion: We have identified a number of regulated genes in T-cells from bone marrow that differed between patients with ITP and healthy controls. The importance of several of the identified genes has already been implicated in ITP, proving the usefulness of this strategy. The identification of novel regulated genes may provide new insights into the physiology and pathophysiology of ITP and for autoimmunity in general. Disclosures: No relevant conflicts of interest to declare.

MS risk genes are transcriptionally regulated in CSF leukocytes at relapse

2012 ◽  
Vol 19 (4) ◽  
pp. 403-410 ◽  
Author(s):  
Margareta Jernås ◽  
Clas Malmeström ◽  
Markus Axelsson ◽  
Caroline Olsson ◽  
Intawat Nookaew ◽  
...  
Keyword(s):  
Gene Expression ◽  
B Cells ◽  
Real Time ◽  
Microarray Analysis ◽  
Dna Microarray ◽  
Nk Cell ◽  
Expression Profiles ◽  
Enzyme Linked Immunosorbent Assay ◽  
Risk Genes ◽  
Dna Microarray Analysis

Background: Infiltrating T-helper cells, cytotoxic T-cells, B-cells and monocytes are thought to mediate the damage to myelin, oligodendrocytes and axons in multiple sclerosis (MS), which results in progressive disability. Objective: The objective of this paper is to explore gene expression profiles of leukocytes in the cerebrospinal fluid (CSF) compartment of MS patients during relapse. Methods: Global gene expression was analyzed by DNA microarray analysis of cells in CSF from MS patients and controls, and verifications were performed with real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Results: Fifty percent of the recently described risk genes for MS and 28% of non-risk genes were differently expressed in MS patients compared to controls (χ2-test, p=7.7 × 10−5). Genes involved in T- and NK-cell processes were up-regulated, and genes involved in processes targeting innate immunity or B-cells were down-regulated in MS. Increased expression of EDN1 and CXCL11 and decreased expression of HMOX1 was verified with real-time PCR and increased expression of CXCL13 was verified with ELISA in CSF. Conclusion: DNA microarray analysis is useful in identifying differently expressed genes in CSF leukocytes, which may be important in MS in vivo. Our findings suggest that many of the risk genes for MS are differently expressed in the disease-mediating leukocytes that penetrate the blood-brain barrier.

scholarly journals POS0785 CHANGING EXPRESSION PROFILES OF LONG NONCODING RNAS, MIRNAS, MRNAS AND CIRCULAR RNAS IN LABIAL SALIVARY GLANDS OF PRIMARY SJÖGREN’S SYNDROME (PSS)

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 645.3-646
Author(s):  
G. Feng ◽  
L. Huang ◽  
J. Ji ◽  
C. Dong ◽  
Y. Xia ◽  
...  
Keyword(s):  
Salivary Glands ◽  
Real Time ◽  
Real Time Pcr ◽  
Expression Profiles ◽  
Sjogren’S Syndrome ◽  
Sjogren's Syndrome ◽  
Primary Sjögren’S Syndrome ◽  
Differentially Expressed ◽  
Primary Sjogren’S Syndrome ◽  
Primary Sjögren's Syndrome

Background:Primary Sjögren’s syndrome (pSS) is a relatively common autoimmune disease characterized by oral and ocular dryness. An increasing number of studies have revealed that long non-coding RNA (lncRNA), miRNA, mRNA and circular RNA (circRNA) contributes to the pathogenesis of autoimmune diseases.Objectives:To explore lncRNA, miRNA, mRNA and circRNA expression profiles in labial salivary glands (LSGs) in pSS patients and their biological functions in the regulation of pSS.Methods:The expression of 75,550 lncRNAs, 2,318 miRNA, 20,292 mRNAs and 6,877 circRNAs were determined in the LSG of six pSS patients and six healthy controls using microarray experiments. Validation was performed in pSS patients and controls using real-time PCR. LncRNA-mRNA co-expression and gene-pathway networks were constructed using bioinformatics software.Results:A total of 599 lncRNAs (upregulated: 279, downregulated: 320), 78 miRNAs (upregulated: 26, downregulated: 52), 615 mRNAs (upregulated: 590, downregulated: 25) and 160 mRNAs (upregulated: 110, downregulated: 50) were differentially expressed in the LSGs of pSS patients. Five of these lncRNAs were validated using real-time PCR. lncRNA HCP5, lncRNA SNHG5, lncRNA IFI44L, lncRNA CMPK2 were significantly upregulated and lncRNA TTYH1 were downregulated in pSS. GO and KEGG biological pathway analysis were performed to predict the functions of differentially expressed lncRNAs and co-expressed potential targeting genes. Subsequently, a ceRNA (lncRNA-miRNA-mRNA) network including 2320 ceRNA pairs was constructed based on predicted miRNAs shared by lncRNAs and mRNAs.Conclusion:The expression profile provided a systematic perspective on the potential functions of lncRNAs miRNAs, mRNAs and circRNAs in the pathogenesis of pSS. Therefore, this study will aid in the development of new diagnostic biomarkers and drug therapies.References:[1]Le Dantec C, Varin MM, Brooks WH, Pers JO, Youinou P, Renaudineau Y. Epigenetics and Sjogren’s syndrome.Curr Pharm Biotechnol. 2012 Aug;13(10):2046-53.Disclosure of Interests:None declared

MicroRNAs Profiling in HIV, HCV and HIV/HCV Coinfected Patients

2020 ◽  
Vol 18 ◽  
Author(s):  
Mohsen Moghoofei ◽  
Sohrab Najafipour ◽  
Shayan Mostafaei ◽  
Ahmad Tavakoli ◽  
Farah Bokharaei-Salim ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Expression Profiles ◽  
Expression Level ◽  
Healthy Controls ◽  
Health Issues ◽  
Viral Loads ◽  
Mirna Expression Pattern ◽  
Immunodeficiency Virus ◽  
Mirna Species

Background: Human immunodeficiency virus (HIV) an hepatitis C virus (HCV) infections are important public health issues. Objective: This study aimed to assess the association between microRNAs expression level and immunological and viral markers in HIV, HCV, and HIV/HCV coinfected patients. Method: The expression level of miR-29, miR-149, miR-199, miR-let7, miR-223, miR-155, miR-122, and miR-150 was evaluated in 20 HIV, 20 HCV, 20 coinfected patients and 20 healthy controls using real-time PCR assay. HIV and HCV viral loads were measured by real-time PCR and also, CD4+ T-lymphocyte count was measured by the PIMA CD4 analyzer. Result: The miRNA expression pattern in each mentioned group showed significantly different expression profiles, but some miRNA species were shared between the groups. MiR-122 and miR-155 were up-regulated while miR-29 and miR-223 were down-regulated in three patients groups compared to healthy controls. A significant positive correlation was observed between the expression of miR-122 and HIV/HCV loads. But, miR-29 and let-7 were negatively correlated with HIV load, and miR149 and let-7 were negatively correlated with HCV load. Also, miR-155 was positively correlated with HCV load. MiR-122 and miR-199 were negatively while others were positively correlated with CD4+ T cell count. Conclusion: These miRNAs are probably involved in the clinical progression and pathogenesis of HIV and HCV infections. Therefore, determining and manipulating of these miRNAs can lead to opening a new gate to control of these important infections.

scholarly journals Real-time DNA microarray analysis

2009 ◽  
Vol 37 (20) ◽  
pp. e132-e132 ◽  
Author(s):  
Arjang Hassibi ◽  
Haris Vikalo ◽  
José Luis Riechmann ◽  
Babak Hassibi
Keyword(s):  
Real Time ◽  
Microarray Analysis ◽  
Dna Microarray ◽  
Dna Microarray Analysis

scholarly journals +3179G/A Insulin-Like Growth Factor-1 Receptor Polymorphism: A Novel Susceptibility Contributor in Anti-Ro/SSA Positive Patients with Sjögren’s Syndrome: Potential Clinical and Pathogenetic Implications

2021 ◽  
Vol 10 (17) ◽  
pp. 3960
Author(s):  
Charalampos Skarlis ◽  
Nikolaos Marketos ◽  
Adrianos Nezos ◽  
Asimina Papanikolaou ◽  
Michael Voulgarelis ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Growth Factor ◽  
Peripheral Blood ◽  
Sjögren's Syndrome ◽  
Sjogren’S Syndrome ◽  
Sjogren's Syndrome ◽  
Genetic Variations ◽  
Healthy Controls ◽  
Insulin Like Growth Factor ◽  
Sjögren‘S Syndrome

Background: Alterations of the insulin-like growth factor (IGF) pathway along with genetic variations of the IGF1 receptor (IGF1R) gene have been linked to the development of systemic autoimmunity, possibly through apoptosis induction. This study aims to investigate whether genetic variations of the IGF1R contribute to Sjögren’s syndrome (SS) pathogenesis and explores potential functional implications. Methods: DNA extracted from whole peripheral blood derived from 277 primary SS patients, complicated or not by lymphoma, and 337 Healthy controls (HC) was genotyped for the rs2229765 IGF1R polymorphism using the RFLP-PCR assay. Gene expression of IGF1R and IGF1 isoforms, caspases 1, 4, and 5, and inflammasome components NLRP3, ASC, IL1β, IL18, IL33, IGFBP3, and IGFBP6 were quantitated by RT-PCR in total RNA extracted from minor salivary gland biopsies (MSGs) of 50 SS patients and 13 sicca controls (SCs). In addition, IGF1R immunohistochemical (IHC) expression was assessed in formalin-fixed, paraffin-embedded MSG tissue sections derived from 10 SS patients and 5 SCs. Results: The prevalence of the A/A genotype of the rs2229765 IGF1R polymorphism was significantly higher in the anti-Ro/SSA positive SS population compared to healthy controls (24.8% vs. 10.7%, p = 0.001). Moreover, IGF1Rs at both mRNA and protein levels were reduced in SS-derived MSGs compared to SCs and were negatively associated with caspase 1 transcripts. The latter were positively correlated with NLRP3, ASC, and IL1β at the salivary gland tissue level. IGF1R expression in peripheral blood was negatively correlated with ESR and IgG serum levels and positively correlated with urine-specific gravity values. Conclusions: The rs2229765 IGF1R variant confers increased susceptibility for seropositive primary SS. Dampened IGF1R mRNA and protein expression in salivary gland tissues could be related to increased apoptosis and subsequently to the activation of inflammasome pathways.

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