Integrating the lncRNA and mRNA expression profiles of TLR4-primed mesenchymal stem cells in ankylosing spondylitis

Abstract Background: Ankylosing spondylitis (AS) is a chronic autoimmune disease, and the precise pathogenesis is largely unknown at present. Our previous study found that the expression of toll-like receptor 4 (TLR4) of mesenchymal stem cells from AS patients (AS-MSCs) was reduced and activation of TLR4 by lipopolysaccharide (LPS) could enhance the immunoregulatory ability of AS-MSCs. However, the potential mechanism by which TLR4 affect the immunoregulatory function of AS-MSCs remains unclear. Objective: The goal of this study was to explore the expression profiles and functional networks of lncRNAs and mRNAs in TLR4-primed AS-MSCs and to clarify the potential mechanisms by which TLR4-primed AS-MSCs exert immunoregulatory effects.Methods: Immunoregulatory effects of MSCs were determined after TLR4 activation. Then, the differentially expressed (DE) long non-coding RNAs (lncRNA) and messenger RNAs (mRNA) between AS-MSCs and TLR4-primed AS-MSCs (stimulated by LPS) were identified through high-throughput sequencing followed by qRT-PCR confirmation. Finally, bioinformatic analyses were performed to identify the critical biological functions, signalling pathways and associated functional networks involved in the TLR4-primed immunoregulatory function of AS-MSCs.Results: A total of 147 DE lncRNAs and 698 DE mRNAs were identified between TLR4-primed AS-MSCs and unstimulated AS-MSCs. Of total, 107 lncRNAs were upregulated and 40 were downregulated (fold change ≥2, P <0.05), while 504 mRNAs upregulated and 194 downregulated (fold change ≥2, P <0.05). 5 lncRNAs and 5 mRNAs with largest fold changes were respectively verified by qRT-PCR. GO and KEGG analysis demonstrated that the DE mRNAs and lncRNAs were highly associated with the inflammatory response, such as NOD-like receptor (NLR) signalling pathway, the TNF signalling pathway and the NF-kappa B signalling pathway. Cis-regulation prediction revealed 8 novel lncRNAs while trans-regulation prediction revealed 15 lncRNAs, respectively. 8 core pairs of LncRNA and target mRNA in the lncRNA-TF-mRNA network were: PACERR-PTGS2, LOC105378085-SOD2, LOC107986655-HIVEP2, MICB-DT-MICB, LOC105373925-SP140L, LOC107984251-IFIT5, LOC112268267-GBP2 and LOC101926887-IFIT3, respectively.Conclusion: In AS, TLR4 activation can enhance the immunoregulation ability of MSCs. Eight core pairs of lncRNA and target mRNA have been found in TLR4-primed AS-MSCs, which could contribute to elucidate the potential mechanism of immunoregulatory dysfunction of AS-MSCs.

Download Full-text

Integrative analysis of long non-coding RNA (lncRNA) and mRNA expression in TLR4-primed MSCs of ankylosing spondylitis

10.21203/rs.3.rs-45893/v2 ◽

2020 ◽

Author(s):

Yuxi Li ◽

Ming Li ◽

Ting Liu ◽

Jiajun Huang ◽

Yuwei Liang ◽

...

Keyword(s):

Ankylosing Spondylitis ◽

High Throughput Sequencing ◽

Expression Profiles ◽

Fold Change ◽

Signalling Pathway ◽

Functional Networks ◽

Potential Mechanism ◽

Target Mrna ◽

Qrt Pcr ◽

Tlr4 Activation

Abstract Background: Ankylosing spondylitis (AS) is a chronic autoimmune disease, and the precise pathogenesis is largely unknown at present. Our previous study found that the expression of toll-like receptor 4 (TLR4) of mesenchymal stem cells from AS patients (AS-MSCs) was reduced and activation of TLR4 by lipopolysaccharide (LPS) could enhance the immunoregulatory ability of AS-MSCs. However, the potential mechanism by which TLR4 affect the immunoregulatory function of AS-MSCs remains unclear. Objective: The goal of this study was to explore the expression profiles and functional networks of lncRNAs and mRNAs in TLR4-primed AS-MSCs and to clarify the potential mechanisms by which TLR4-primed AS-MSCs exert immunoregulatory effects.Methods: Immunoregulatory effects of MSCs were determined after TLR4 activation. Then, the differentially expressed (DE) long non-coding RNAs (lncRNA) and messenger RNAs (mRNA) between AS-MSCs and TLR4-primed AS-MSCs (stimulated by LPS) were identified through high-throughput sequencing followed by qRT-PCR confirmation. Finally, bioinformatic analyses were performed to identify the critical biological functions, signalling pathways and associated functional networks involved in the TLR4-primed immunoregulatory function of AS-MSCs.Results: A total of 147 DE lncRNAs and 698 DE mRNAs were identified between TLR4-primed AS-MSCs and unstimulated AS-MSCs. Of total, 107 lncRNAs were upregulated and 40 were downregulated (fold change ≥2, P <0.05), while 504 mRNAs upregulated and 194 downregulated (fold change ≥2, P <0.05). 5 lncRNAs and 5 mRNAs with largest fold changes were respectively verified by qRT-PCR. GO and KEGG analysis demonstrated that the DE mRNAs and lncRNAs were highly associated with the inflammatory response, such as NOD-like receptor (NLR) signalling pathway, the TNF signalling pathway and the NF-kappa B signalling pathway. Cis-regulation prediction revealed 8 novel lncRNAs while trans-regulation prediction revealed 15 lncRNAs, respectively. 8 core pairs of LncRNA and target mRNA in the lncRNA-TF-mRNA network were: PACERR-PTGS2, LOC105378085-SOD2, LOC107986655-HIVEP2, MICB-DT-MICB, LOC105373925-SP140L, LOC107984251-IFIT5, LOC112268267-GBP2 and LOC101926887-IFIT3, respectively.Conclusion: In AS, TLR4 activation can enhance the immunoregulation ability of MSCs. Eight core pairs of lncRNA and target mRNA have been found in TLR4-primed AS-MSCs, which could contribute to elucidate the potential mechanism of immunoregulatory dysfunction of AS-MSCs.

Download Full-text

Gene Expression of Adhesion Molecules in Endothelial Cells from Patients with Peripheral Arterial Disease Is Reduced after Surgical Revascularization and Pharmacological Treatment

International Journal of Vascular Medicine ◽

10.1155/2013/412761 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 4

Author(s):

Franca Marino ◽

Luigina Guasti ◽

Matteo Tozzi ◽

Laura Schembri ◽

Luana Castiglioni ◽

...

Keyword(s):

Gene Expression ◽

Endothelial Cells ◽

Mrna Expression ◽

Adhesion Molecules ◽

Mononuclear Cells ◽

Venous Blood ◽

Statin Treatment ◽

Early Event ◽

Mrna Levels ◽

Peripheral Blood Mononuclear

Atherosclerosis is an inflammatory disease characterized by immunological activity, in which endothelial dysfunction represents an early event leading to subsequent inflammatory vascular damage. We investigated gene expression of the adhesion molecules (AMs) ICAM-1, VCAM-1, andβ1-integrin in endothelial cells (ECs) isolated from venous blood (circulating EC, cEC) and purified from femoral plaques (pEC) obtained from 9 patients with peripheral artery disease (PAD) submitted to femoral artery thrombendarterectomy (FEA). In addition, in peripheral blood mononuclear cells (PBMCs) of the same subjects, we investigated gene expression of IFN-γ, IL-4, TGF-β, and IL-10. Patients were longitudinally evaluated 1 month before surgery, when statin treatment was established, at the time of surgery, and after 2 and 5 months. All AM mRNA levels, measured by means of real-time PCR, in cEC diminished during the study, up to 41–50% of initial levels at followup. AM mRNA expression was significantly higher in pEC than in cEC. During the study, in PBMCs, TGF-βand IL-10 mRNA levels remained unchanged while IFN-γand IL-4 levels increased; however, the ratio IFN-γ/IL-4 showed no significant modification. In PAD patients, FEA and statin treatment induce a profound reduction of AM expression in cEC and affect cytokine mRNA expression in PBMCs.

Download Full-text

Circular RNA expression profiles and bioinformatic analysis in coronary heart disease

Epigenomics ◽

10.2217/epi-2019-0369 ◽

2020 ◽

Vol 12 (5) ◽

pp. 439-454 ◽

Cited By ~ 2

Author(s):

Fangpu Yu ◽

Yuanyuan Tie ◽

Ya Zhang ◽

Zunzhe Wang ◽

Liwen Yu ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Coronary Arteries ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Expression Profiles ◽

Peripheral Blood Mononuclear ◽

Qrt Pcr ◽

Blood Mononuclear Cells

Aim: We aimed to identify the expression profile and role of circular RNAs (circRNAs) in coronary heart disease (CHD). Materials & methods: We performed sequence analysis of circRNAs in peripheral blood mononuclear cells of 70 CHD patients and 30 controls. Eight selected circRNAs were validated using quantitative real-time polymerase chain reaction (qRT-PCR) in human atherosclerotic coronary arteries. Results: In total, 2283 downregulated and 85 upregulated circRNAs were identified in CHD. Parental genes of top 100 dysregulated-circRNAs are related to metabolism and protein modification, and 12 circRNAs might upregulate their CHD-related parental genes through miRNA sponges. Of the eight circRNAs validated in atherosclerotic coronary arteries by qRT-PCR, six were consistent with sequencing results of peripheral blood mononuclear cells. Conclusion: As potential ceRNAs, dysregulated circRNAs may be involved in CHD pathophysiology.

Download Full-text

DDAH2 mRNA Expression Is Inversely Associated with Some Cardiovascular Risk-Related Features in Healthy Young Adults

Disease Markers ◽

10.1155/2009/978157 ◽

2009 ◽

Vol 27 (1) ◽

pp. 37-44 ◽

Cited By ~ 6

Author(s):

Blanca Puchau ◽

Helen Hermana M. Hermsdorff ◽

M. Ángeles Zulet ◽

J. Alfredo Martínez

Keyword(s):

Young Adults ◽

Cardiovascular Risk ◽

Mrna Expression ◽

Inflammatory Markers ◽

Mononuclear Cells ◽

Expression Profiles ◽

Peripheral Blood Mononuclear ◽

Disease Marker ◽

Reduced Risk ◽

Apparently Healthy

The purpose of this study was to evaluate whether the mRNA expression profiles of three genes (PRMT1, DDAH2 and NOS3) are related to ADMA metabolism and signalling, and the potential relationships with anthropometrical, biochemical, lifestyle and inflammatory indicators in healthy young adults. An emphasis on the putative effect of different mRNA expression on cardiovascular risk-related features was paid. Anthropometrical measurements as well as lifestyle features were analyzed in 120 healthy young adults. Fasting blood samples were collected for the measurement of glucose and lipid profiles as well as the concentrations of selected inflammatory markers. Profiles of mRNA expression were assessed for PRMT1, DDAH2 and NOS3 genes from peripheral blood mononuclear cells. Regarding inflammatory biomarkers, DDAH2 was inversely associated with IL-6 and TNF-α. Moreover, subjects in the highest quintile of DDAH2 mRNA expression showed a reduced risk to have higher values of waist circumference, and to be more prone to show higher values of HDL-c. Interestingly, DDAH2 gene expression seemed to be related with some anthropometrical, biochemical, lifestyle and inflammatory indicators linked to cardiovascular risk in apparently healthy young adults, emerging as a potential disease marker.

Download Full-text

Tonsils of the Soft Palate Do Not Mediate the Response of Pigs to Oral Vaccination with Heat-Inactivated Mycobacterium bovis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00221-14 ◽

2014 ◽

Vol 21 (8) ◽

pp. 1128-1136 ◽

Cited By ~ 8

Author(s):

Beatriz Beltrán-Beck ◽

Beatriz Romero ◽

Mariana Boadella ◽

Carmen Casal ◽

Javier Bezos ◽

...

Keyword(s):

Wild Boar ◽

Mycobacterium Bovis ◽

Mononuclear Cells ◽

Mammalian Species ◽

Serum Antibody ◽

Control Group ◽

Mrna Levels ◽

Oral Vaccination ◽

Peripheral Blood Mononuclear ◽

Content Type

ABSTRACTMycobacterium boviscauses animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivatedM. bovisand challenge with a virulentM. bovisfield strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests forin vivoTB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivatedM. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. MassiveM. bovisgrowth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased afterM. bovischallenge. This information is relevant for pig production in regions that are endemic forM. bovisand for TB vaccine research.

Download Full-text

Serum Interferon (IFN)-Neutralizing Antibodies and Bioactivities of IFNs in Patients with Severe Type II Essential Mixed Cryoglobulinemia

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.1.70-77.2003 ◽

2003 ◽

Vol 10 (1) ◽

pp. 70-77 ◽

Cited By ~ 6

Author(s):

Carolina Scagnolari ◽

Milvia Casato ◽

Francesca Bellomi ◽

Francesca De Pisa ◽

Ombretta Turriziani ◽

...

Keyword(s):

Mrna Expression ◽

Clinical Response ◽

Neutralizing Antibodies ◽

Mononuclear Cells ◽

Mixed Cryoglobulinemia ◽

Mrna Levels ◽

Type Ii ◽

Essential Mixed Cryoglobulinemia ◽

Peripheral Blood Mononuclear ◽

Severe Type

ABSTRACT The efficacy of alpha interferon (IFN-α) in the treatment of severe type II essential mixed cryoglobulinemia (EMC) has been reported previously. In some patients, the development of neutralizing antibodies to recombinant IFN-α (rIFN-α) can affect the clinical response achieved with rIFN-α; a second treatment with natural IFN-α preparations may reinduce the clinical response. In the present study the ability of leukocyte IFN (LeIFN) to restore the response was investigated from a pharmacodynamic viewpoint. Specifically, the pharmacodynamic profiles of different IFN-α preparations were studied by measuring the serum neopterin levels and the levels of expression of protein MxA mRNA in in vivo peripheral blood mononuclear cells in two patients with EMC whose resistance to rIFN-α2a treatment increased concomitantly with the development of neutralizing antibodies. These markers were measured before injection and at 24 and 48 h after a single injection of rIFN-α2a, consensus IFN [(C)IFN], or LeIFN. No increase or only a slight increase in MxA mRNA levels was detectable after administration of rIFN-α2a or (C)IFN, whereas a significant increase (≥10-fold) in MxA mRNA expression was recorded following administration of LeIFN. The neutralizing antibodies to rIFN-α2a cross-react with (C)IFN. Sera from these patients neutralized most but not all of the subtypes present in the natural IFN-α (LeIFN) mixture, and no significant increase in neopterin levels was observed after these patients were switched to LeIFN treatment. In summary, the data demonstrate that the problem of neutralizing antibodies still exists and that LeIFN may induce an increase in the level of MxA mRNA expression but not an increase in neopterin levels in patients who are resistant to treatment with rIFN-α2a or (C)IFN.

Download Full-text

Elevated Expression of the NLRP3 Inflammasome and Its Correlation with Disease Activity in Adult-onset Still Disease

The Journal of Rheumatology ◽

10.3899/jrheum.161354 ◽

2017 ◽

Vol 44 (8) ◽

pp. 1142-1150 ◽

Cited By ~ 17

Author(s):

Chia-Wei Hsieh ◽

Yi-Ming Chen ◽

Chi-Chen Lin ◽

Kuo-Tung Tang ◽

Hsin-Hua Chen ◽

...

Keyword(s):

Disease Activity ◽

Mrna Expression ◽

Nlrp3 Inflammasome ◽

Mononuclear Cells ◽

Mrna Levels ◽

Adult Onset ◽

Peripheral Blood Mononuclear ◽

Protein Expressions ◽

Still Disease ◽

Stimulation Of

Objective.The dysregulation of the NLRP3 (NLR containing a pyrin domain) inflammasome is involved in autoinflammatory diseases. Adult-onset Still disease (AOSD) is regarded as an autoinflammatory disease. However, the pathogenic involvement of NLRP3 inflammasome in AOSD remains unclear and NLRP3 activators in AOSD are currently unknown.Methods.The mRNA expression of NLRP3 inflammasome signaling in peripheral blood mononuclear cells (PBMC) from 34 patients with AOSD and 14 healthy subjects was determined using quantitative-PCR (qPCR). The changes in mRNA and protein levels of NLRP3 inflammasome signaling in PBMC treated with the potential activator [imiquimod (IMQ)] or inhibitor of NLRP3 were evaluated using qPCR and immunoblotting, respectively. The supernatant levels of interleukin (IL)-1β and IL-18 were determined by ELISA.Results.Significantly higher mRNA levels of NLRP3 inflammasome signaling were observed in patients with AOSD compared with healthy controls. NLRP3 expressions were positively correlated with disease activity in patients with AOSD. IMQ (an effective Toll-like receptor 7 ligand; 10 µg/ml and 25 µg/ml) stimulation of PBMC from patients with AOSD induced dose-dependent increases of mRNA expression of NLRP3 (mean ± standard error of the mean, 2.06 ± 0.46 and 6.05 ± 1.84, respectively), caspase-1 (1.81 ± 0.23 and 4.25 ± 0.48), IL-1β (5.68 ± 1.51 and 12.13 ± 3.71), and IL-18 (2.32 ± 0.37 and 4.81 ± 0.51) compared with controls (all p < 0.005). IMQ stimulation of PBMC from patients similarly induced greater increases in protein expressions of NLRP3 inflammasome compared with controls. The protein expressions of NLRP3, IL-1β, and IL-18 on PBMC significantly decreased after treatment with NLRP3 inhibitor in patients with AOSD.Conclusion.Increased expression of NLRP3 inflammasome and its positive correlation with disease activity in AOSD suggest its involvement in disease pathogenesis. IMQ upregulated expressions of NLRP3 inflammasome signaling, and IMQ might be an activator of NLRP3 inflammasome in AOSD.

Download Full-text

Celiac Disease Causes Epithelial Disruption and Regulatory T Cell Recruitment in the Oral Mucosa

Frontiers in Immunology ◽

10.3389/fimmu.2021.623805 ◽

2021 ◽

Vol 12 ◽

Author(s):

Javier Sanchez-Solares ◽

Luis Sanchez ◽

Carmela Pablo-Torres ◽

Celso Diaz-Fernandez ◽

Poul Sørensen ◽

...

Keyword(s):

Celiac Disease ◽

Oral Mucosa ◽

Mononuclear Cells ◽

De Novo ◽

Current Treatment ◽

Oral Immunotherapy ◽

Control Group ◽

Mrna Levels ◽

Peripheral Blood Mononuclear ◽

Diagnostic Potential

Celiac disease (CD) is a chronic autoimmune disease characterized by an immune-triggered enteropathy upon gluten intake. The only current treatment available is lifelong Gluten Free Diet (GFD). Several extraintestinal manifestations have been described in CD, some affecting the oral mucosa. Thus, we hypothesized that oral mucosa could potentially be a target for novel biomarkers and an administration route for CD treatment. Six de novo diagnosed and seven CD patients under GFD for at least 1 year were recruited. Non-celiac subjects (n = 8) were recruited as control group. Two biopsies of the cheek lining were taken from each subject for mRNA analysis and immunohistochemical characterization. We observed a significant decrease in the expression of epithelial junction proteins in all CD patients, indicating that oral mucosa barrier integrity is compromised. FoxP3+ population was greatly increased in CD patients, suggesting that Tregs are recruited to the damaged mucosa, even after avoidance of gluten. Amphiregulin mRNA levels from Peripheral Blood Mononuclear Cells (PBMCs) and epithelial damage in the oral mucosa correlated with Treg infiltration in all the experimental groups, suggesting that recruited Tregs might display a “repair” phenotype. Based on these results, we propose that oral mucosa is altered in CD and, as such, might have diagnostic potential. Furthermore, due to its tolerogenic nature, it could be an important target for oral immunotherapy.

Download Full-text

Serum urate is related to subclinical inflammation in asymptomatic hyperuricaemia

Rheumatology ◽

10.1093/rheumatology/keaa425 ◽

2020 ◽

Vol 60 (1) ◽

pp. 371-379 ◽

Cited By ~ 1

Author(s):

Desirée Luis-Rodríguez ◽

Javier Donate-Correa ◽

Ernesto Martín-Núñez ◽

Carla Ferri ◽

Víctor G Tagua ◽

...

Keyword(s):

Mrna Expression ◽

Mononuclear Cells ◽

High Sensitivity ◽

Serum Levels ◽

Normal Serum ◽

Control Group ◽

Subclinical Inflammation ◽

Peripheral Blood Mononuclear ◽

Tnf Α ◽

Inflammatory Profile

Abstract Objective Asymptomatic hyperuricaemia (AHU) is associated with inflammatory disorders, including cardiovascular disease. Uric acid (UA) lowering therapies may reduce the risk of appearance or the progression of these comorbidities. In this work, we investigated the relationship between serum UA levels and inflammation in subjects with AHU. Methods Serum levels of high-sensitivity CRP (hsCRP), TNF-α and IL-6, and mRNA expression of TNFa and IL6 in peripheral blood mononuclear cells were measured in individuals with AHU and without comorbid conditions and in a control group with similar characteristics and normal serum UA levels. Additionally, we determined the variations in the inflammatory profile in a subgroup of subjects after 6 months of treatment with allopurinol. Results Subjects at higher tertiles of serum UA presented higher levels of hsCRP and increased serum and mRNA expression levels of both cytokines (P < 0.001). UA levels constituted an independent predictor of increased levels of inflammatory parameters in multiple regression models (P < 0.001) and a risk factor for the presence of a subclinical inflammation in multivariate logistic regression (P < 0.001). Allopurinol reduced UA and serum and mRNA expression of inflammatory cytokines (P < 0.001). There was a significant correlation between the variations in serum UA and the variations in serum TNF-α (P < 0.01) and IL-6 (P < 0.05), and mRNA expression of these cytokines (P < 0.05). This association remained significant and independent (P < 0.01). Conclusion In subjects with AHU, serum UA may be an inductor of subclinical inflammation. Therapeutic reduction of serum UA was associated with a modulation of the inflammatory profile.

Download Full-text