A Comparative Study on the Incidence, Aggravation, and Remission of Lupus Nephritis Based on iTRAQ Technology

Aim and Objective: Lupus nephritis (LN) is one of the major complications of systemic lupus erythematosus (SLE). The specific mechanisms of pathogenesis, aggravation, and remission processes in LN have not been clarified but is of great need in the clinic. Using isobaric tags for relative and absolute quantitation (iTRAQ) technology to screen the functional proteins of LN in mice. Especially under intervention factors of lipopolysaccharide (LPS) and dexamethasone. Methods: Mrl-lps mice were intervened with LPS, dexamethasone, and normal saline (NS) using intraperitoneal injection, and c57 mice intervened with NS as control. The anti-ANA antibody enzyme-linked immunosorbent assay (ELISA) was used to verify disease severity. Kidney tissue is collected and processed for iTRAQ to screen out functional proteins closely related to the onset and development of LN. Western blot method and rt-PCR (real-time Polymerase Chain Reaction) were used for verification. Results: We identified 136 proteins that marked quantitative information. Among them, Hp, Igkv8-27, Itgb2, Got2, and Pcx proteins showed significant abnormal manifestations. Conclusion: Using iTRAQ methods, the functional proteins Hp, Igkv8-27, Itgb2, Got2, and Pcx were screened out for a close relationship with the pathogenesis and development of LN, which is worth further study.

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Lipidomics Revealed Aberrant Metabolism of Lipids Including FAHFAs in Renal Tissue in the Progression of Lupus Nephritis in a Murine Model

Metabolites ◽

10.3390/metabo11030142 ◽

2021 ◽

Vol 11 (3) ◽

pp. 142

Author(s):

Changfeng Hu ◽

Yu Du ◽

Xiaofen Xu ◽

Haichang Li ◽

Qiao Duan ◽

...

Keyword(s):

Oxidative Stress ◽

Fatty Acid ◽

Lupus Nephritis ◽

Lupus Erythematosus ◽

Kidney Tissue ◽

Renal Tissue ◽

Autoimmune Response ◽

Enzyme Linked Immunosorbent Assay ◽

Shotgun Lipidomics ◽

Necrosis Factor Alpha

Lupus nephritis (LN) is an inflammatory renal disease of patients with systemic lupus erythematosus with lots of immune complexes deposited in kidneys. Accumulated studies have demonstrated the close relationships among dyslipidaemia, inflammation, and autoimmune response, and oxidative stress in the patients. Lipids play numerous important roles in biological process and cellular functions. Herein, shotgun lipidomics was employed to quantitatively analyze cellular lipidomes in the renal tissue of MRL/lpr mice in the progression of LN (including pre-LN and LN state) with/without treated with glucocorticoids (GCs). The levels of cytokines (i.e., TNF-α (Tumor necrosis factor alpha) and IL-6 (Interleukin 6)) in the serum were measured by ELISA (enzyme-linked immunosorbent assay) kits. Renal histopathological changes and C3 deposition in the glomeruli of the mice were also determined. Lipidomics analysis revealed that the ectopic fat deposition and the aberrant metabolism of lipids that were relevant to oxidative stress (e.g., 4-hydroxyalkenal, ceramide, lysophospholipid species, etc.) always existed in the development of LN. Moreover, the anti-inflammatory FAHFA (fatty acid ester of hydroxyl fatty acid) species in the kidney tissue could largely reflect the severity of LN. Thus, they were a potential early biomarker for LN. In addition, the study also revealed that treatment with GCs could prevent the progression of LN, but greatly aggravate the aberrant metabolism of the lipids, particularly when used for a long time.

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Correlation Between Serological Makers and Immunofluorescence Deposits i n Kidney Tissue of Patients with Lupus Nephritis

International Journal of Drug Delivery Technology ◽

10.25258/ijddt.9.2.22 ◽

2019 ◽

Vol 9 (02) ◽

Author(s):

Haider S Al-Hadad ◽

Aqeel Abbas Matrood ◽

Maha Abdalrasool Almukhtar ◽

Haider Jabur Kehiosh ◽

Riyadh Muhi Al-Saegh

Keyword(s):

Lupus Nephritis ◽

Lupus Erythematosus ◽

Kidney Biopsy ◽

Pearson Correlation ◽

Standard Procedure ◽

Kidney Tissue ◽

P Value ◽

American College Of Rheumatology ◽

Immune Deposits ◽

Number Of Patients

Background: Systemic lupus erythematosus (SLE) is an autoimmune disease. Few biomarkers for SLE have been validated and widely accepted for the laboratory follow-up of inflammatory activity. In SLE patients, with lupus nephritis (LN), complement activation leads to fluctuation of serum C3 and C4 that are frequently used as clinicalm biomarker of disease activity in SLE. Patients and Methods: In this study the number of patients were 37, seven patients were excluded for incomplete data collection, 28 were females ,2 were males. The duration of the study is two years from 2015 to 2017. Patients were considered to have SLE and LN according to American College of Rheumatology (ACR) criteria, and International Society of Nephrology/ Renal Pathology Society (ISN/RPS). All patients were evaluated withm clinical presentation, laboratory investigations. Our patients underwent kidney biopsy according to standard procedure by Kerstin Amann, and their tissue specimens were studied in the laboratory with light microscope (LM) and immunofluorescence microscope reagents. The relationship between the serological markers and immunofluorescence deposits in kidney biopsy of all patients were studied using the statistical analysis of Pearson correlation and single table student's T test. A P value 0.05 was considered statistically significant. Results: The granular pattern of IF deposits was present in all LN patients, and in more than two third of patients these IF deposits presented in glomerular, tubular, and mesangium sites. While less than one third of patients had IF deposits in the mesangium only. There was no statistically significant correlation between serum ANA, anti-dsDNA, and IF deposits of different types. There was significant correlation between serum C3 and C4 hypocomplementemia and IgG immune deposits in kidney biopsy, and there was significant relationship between serum C3 hypocomplementemia and full house immunofluorescence (FHIF) deposits inm kidney biopsy.Conclusions:Immunofluorescence deposits is mainly granular pattern in LN patients. There was no significant association between serum ANA, anti-dsDNA, and immune deposits in kidney tissue. Immunofluorescence deposits of IgG type correlates significantly with serum C3 and C4 hypocomplemetemia, and these immune deposits in association with low complement levels correlates with LN flare. There was significant correlation between C3 hypocomplementemia and FHIF.

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Validity test of anti-c1q serum as diagnostic marker for lupus nephritis

Indonesian Journal of Rheumatology ◽

10.37275/ijr.v8i1.8 ◽

2018 ◽

Vol 8 (1) ◽

Author(s):

M Enrica ◽

A Tjandrawati ◽

S Rachmayati ◽

Laniyati Hamijoyo

Keyword(s):

Systemic Lupus Erythematosus ◽

Lupus Nephritis ◽

Lupus Erythematosus ◽

Predictive Value ◽

Renal Involvement ◽

Enzyme Linked Immunosorbent Assay ◽

Specific Marker ◽

Systemic Lupus ◽

Urine Protein ◽

American College Of Rheumatology

Background: Lupus nephritis is defined as renal involvement in systemic lupus erythematosus (SLE) patients and the most important cause of morbidity and mortality. The diagnostic criteria that used to diagnose lupus nephritis are 1997 American Collegeof Rheumatology is 24 hours urine protein ≥500 mg and/or cellular cast, but significant renal damage can occur without proteinuria or cellular cast. Anti-C1q is an autoantibody that is produced by a chronic alteration of C1q collagen domain. Anti-C1q is a new specific marker for renal marker.Objective: To determine the validity of anti-C1q serum by using 1997 American College of Rheumatology criteria as a gold standard. Methods: This is a cross sectional study, conducted in October to December 2014 at Hasan Sadikin Hospital Bandung. The subjects had systemic lupus erythematosus with and without renal involvement, based on 1997 American College of Rheumatology criteria for SLE.Results: There were 65 subjects included in this study, 64 subjects were female and 1 subject was male. The age average was 32 (SD 11.7) years old. As many as 66.2% subjects had been diagnosed with lupus erythematosus systemic at least 3 years. Twenty four hours urine protein was measured using spectrophotometry, urine sediment was examinedfor cellular cast, and anti-C1q serum was measured using micro enzyme linked immunosorbent assay. Based on American College of Rheumatology criteria, 34 subjects were classified as lupus nephritis group while 31 subjects were classified as non-lupus nephritis group. The area under the curve of anti-C1q was 0.610. The cut-off value used in this study was 10.43 U/ml. The sensitivity, specificity, positive predictive value,negative predictive value and accuracy of anti-C1q assay were 41.18%, 77.42%, 66.67%, 54.55% and 58.46% respectively.Conclusion: Anti-C1q assay, based on this study, hasa low sensitivity and medium specificity to detect lupusnephritis

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Novel Effects of Combination Therapy Through Inhibition of Caspase-1/Gasdermin D Induced-Pyroptosis in Lupus Nephritis

Frontiers in Immunology ◽

10.3389/fimmu.2021.720877 ◽

2021 ◽

Vol 12 ◽

Author(s):

Heng Cao ◽

Junyu Liang ◽

Jing Liu ◽

Ye He ◽

Yini Ke ◽

...

Keyword(s):

Complete Remission ◽

Lupus Nephritis ◽

Combination Therapy ◽

Disease Progression ◽

Lupus Erythematosus ◽

Peripheral Blood ◽

Activity Index ◽

Kidney Tissue ◽

Cell Numbers ◽

Caspase 1

ObjectivesCombination therapy with mycophenolate mofetil, tacrolimus and steroids are effective in achieving complete remission in lupus nephritis (LN). Combination therapy uniquely downregulated caspase-1 compared with monotherapies, which can cleave gasdermin D (GSDMD) and was recently identified as the pyroptosis executioner. We therefore investigated whether combination therapy enabled the suppression of caspase-1/GSDMD-mediated pyroptosis in LN.MethodsExpression and activation of GSDMD were detected in kidney specimens of the human and mouse with LN using immunohistochemical staining and immunoblotting. Primary podocytes isolated from MRL/lpr mice were incubated with LPS+ATP, and pretreated with monotherapy or combination therapy. Inhibition of caspase-1/GSDMD-induced pyroptosis by combination therapy were assessed in MRL/lpr mice and human specimens. Pyroptosis was examined using a FAM caspase-1 kit and flow cytometry. The correlation between pyroptosis in peripheral blood and the systemic lupus erythematosus disease activity index (SLEDAI) was analyzed.ResultsKidney tissue specimens from LN patients and mice exhibited greatly increased expression levels and cleavage of GSDMD. In cultured podocytes, combination treatment significantly suppressed the activation of NLRP3 and caspase-1 and reduced GSDMD N-terminal levels. Combination therapy repressed disease progression through inhibition of caspase-1/GSDMD-mediated pyroptosis in both humans and MRL/lpr mice. Caspase-1/PI positive cell numbers in peripheral blood were positively correlated with SLE-DAI. LN patients with complete remission and partial remission had remarkably reduced caspase-1/PI positive cell numbers compared to baseline. Ac-FLTD-CMK, a GSDMD-derived inhibitor, prevented the development of LN.ConclusionCombination therapy suppressed caspase-1/GSDMD-mediated pyroptosis in vitro and in vivo and reduced disease progression.

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Quantitative Proteomics Analysis of Differentially Expressed Proteins in Serum of Former Uranium Miners by Isobaric Tags for the Relative and Absolute Quantitation

Dose-Response ◽

10.1177/15593258211056190 ◽

2021 ◽

Vol 19 (4) ◽

pp. 155932582110561

Author(s):

Xuhong Dang ◽

Haipeng Lin ◽

Yayi Yuan ◽

Biao Yang ◽

Juancong Dong ◽

...

Keyword(s):

Radiation Damage ◽

Process Analysis ◽

Density Lipoprotein ◽

Differentially Expressed ◽

Enzyme Linked Immunosorbent Assay ◽

Differentially Expressed Proteins ◽

Serum Samples ◽

Absolute Quantitation ◽

Uranium Miners ◽

Isobaric Tags

The carcinogenicity of radon has been convincingly documented through epidemiological studies of underground miners. However, there is a lack of early warning indicators for radon radiation damage. In this study, mixed serum samples of 3 groups were collected from 27 underground uranium miners and seven aboveground miners according to the radiation exposure dose. The differentially expressed proteins in the serum were identified using the isobaric tags for the relative and absolute quantitation (iTRAQ)-based method. Some differentially expressed proteins were validated by enzyme-linked immunosorbent assay (ELISA) in 84 underground and 32 aboveground miners. A total of 25 co-differentially expressed proteins in 2 underground miner groups were screened, of which 9 were downregulated and 13 were upregulated. Biological process analysis of these proteins using Metascape showed that 5 GO terms were enriched, such as negative regulation of very-low-density lipoprotein particle clearance, endocytosis, and regulated exocytosis. The results of the ELISA for the expression levels of GCN1, CIP2A, and IGHV1-24 in the serum of 116 miners’ serum showed that the levels of GCN1 and CIP2A were consistent with the iTRAQ results. In conclusion, APOC1, APOC2, APOC3, ORM1, ORM2, ANTXR1, GCN1, and CIP2A may be potential early markers of radon radiation damage.

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Kidney tissue hypoxia dictates T cell–mediated injury in murine lupus nephritis

Science Translational Medicine ◽

10.1126/scitranslmed.aay1620 ◽

2020 ◽

Vol 12 (538) ◽

pp. eaay1620 ◽

Cited By ~ 7

Author(s):

Ping-Min Chen ◽

Parker C. Wilson ◽

Justin A. Shyer ◽

Margaret Veselits ◽

Holly R. Steach ◽

...

Keyword(s):

T Cells ◽

Lupus Nephritis ◽

Lupus Erythematosus ◽

Hypoxia Inducible Factor ◽

Kidney Tissue ◽

Renal Tissue ◽

Tissue Hypoxia ◽

Mouse Tissue ◽

Low Oxygen ◽

Environmental Adaptations

The kidney is a frequent target of autoimmune injury, including in systemic lupus erythematosus; however, how immune cells adapt to kidney’s unique environment and contribute to tissue damage is unknown. We found that renal tissue, which normally has low oxygen tension, becomes more hypoxic in lupus nephritis. In the injured mouse tissue, renal-infiltrating CD4+ and CD8+ T cells express hypoxia-inducible factor–1 (HIF-1), which alters their cellular metabolism and prevents their apoptosis in hypoxia. HIF-1–dependent gene-regulated pathways were also up-regulated in renal-infiltrating T cells in human lupus nephritis. Perturbation of these environmental adaptations by selective HIF-1 blockade inhibited infiltrating T cells and reversed tissue hypoxia and injury in murine models of lupus. The results suggest that targeting HIF-1 might be effective for treating renal injury in autoimmune diseases.

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DEK Is a Potential Biomarker Associated with Malignant Phenotype in Gastric Cancer Tissues and Plasma

International Journal of Molecular Sciences ◽

10.3390/ijms20225689 ◽

2019 ◽

Vol 20 (22) ◽

pp. 5689 ◽

Cited By ~ 1

Author(s):

Kam-Fai Lee ◽

Ming-Ming Tsai ◽

Chung-Ying Tsai ◽

Chung-Guei Huang ◽

Yu-Hsiang Ou ◽

...

Keyword(s):

Gastric Cancer ◽

Carbohydrate Antigen ◽

Enzyme Linked Immunosorbent Assay ◽

Absolute Quantitation ◽

Roc Curve Analysis ◽

Diagnosis And Prognosis ◽

Potential Biomarker ◽

Ca 19.9 ◽

Novel Biomarkers ◽

Isobaric Tags

Gastric cancer (GC) is the second most widespread cause of cancer-related mortality worldwide. The discovery of novel biomarkers of oncoproteins can facilitate the development of therapeutic strategies for GC treatment. In this study, we identified novel biomarkers by integrating isobaric tags for relative and absolute quantitation (iTRAQ), a human plasma proteome database, and public Oncomine datasets to search for aberrantly expressed oncogene-associated proteins in GC tissues and plasma. One of the most significantly upregulated biomarkers, DEK, was selected and its expression validated. Our immunohistochemistry (IHC) (n = 92) and quantitative real-time polymerase chain reaction (qRT-PCR) (n = 72) analyses disclosed a marked increase in DEK expression in tumor tissue, compared with paired nontumor mucosa. Importantly, significantly higher preoperative plasma DEK levels were detected in GC patients than in healthy controls via enzyme-linked immunosorbent assay (ELISA). In clinicopathological analysis, higher expression of DEK in both tissue and plasma was significantly associated with advanced stage and poorer survival outcomes of GC patients. Data from receiver operating characteristic (ROC) curve analysis disclosed a better diagnostic accuracy of plasma DEK than carcinoembryonic antigen (CEA), carbohydrate antigen 19.9 (CA 19.9), and C-reactive protein (CRP), highlighting its potential as an effective plasma biomarker for GC. Plasma DEK is also more sensitive in tumor detection than the other three biomarkers. Knockdown of DEK resulted in inhibition of GC cell migration via a mechanism involving modulation of matrix metalloproteinase MMP-2/MMP-9 level and vice versa. Our results collectively support plasma DEK as a useful biomarker for making diagnosis and prognosis of GC patients.

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Anti-Double-Stranded DNA Isotypes and Anti-C1q Antibody Improve the Diagnostic Specificity of Systemic Lupus Erythematosus

Disease Markers ◽

10.1155/2018/4528547 ◽

2018 ◽

Vol 2018 ◽

pp. 1-7 ◽

Cited By ~ 4

Author(s):

Yu Jia ◽

Lingling Zhao ◽

Chunyan Wang ◽

Jin Shang ◽

Yi Miao ◽

...

Keyword(s):

Systemic Lupus Erythematosus ◽

Lupus Nephritis ◽

Disease Activity ◽

Lupus Erythematosus ◽

Activity Index ◽

Renal Involvement ◽

Enzyme Linked Immunosorbent Assay ◽

Diagnostic Specificity ◽

Systemic Lupus ◽

Double Stranded Dna

Objectives. We aimed to evaluate the value of immunoglobulin (Ig) G, IgM, and IgA isotypes of anti-double-stranded DNA (anti-dsDNA) and anti-C1q antibody in diagnosing systemic lupus erythematosus (SLE) patients and elucidate their association with disease activity and lupus nephritis. Methods. Blood samples were obtained from 96 SLE patients, 62 other autoimmune disease patients, and 60 healthy blood donors. Anti-dsDNA IgG, IgM, and IgA isotypes and anti-C1q antibody were measured by enzyme-linked immunosorbent assay. Disease activity of SLE patients was assessed according to the SLE Disease Activity Index score. Results. When specificity was greater than 90%, the sensitivity of anti-dsDNA IgG, IgM, and IgA isotypes and anti-C1q antibody in diagnosing SLE was 75%, 45%, 33%, and 49%, respectively. The prevalence of anti-dsDNA IgG (p=0.002), anti-dsDNA IgA (p=0.028), and anti-C1q antibody (p=0.000) in active cases was significantly higher than those in inactive ones. In addition, the presence of anti-C1q antibody was associated with renal involvement (p=0.032). Anti-dsDNA IgM showed no significant association with disease activity, but it was inversely linked with lupus nephritis (p=0.005). When anti-dsDNA IgG and IgA and anti-C1q were combined to evaluate SLE disease activity, the specificity reached the highest level (90%). When anti-C1q positive was accompanied by anti-dsDNA IgM negative, the specificity of diagnosing lupus nephritis was up to 96%. Conclusions. This study demonstrated the role of anti-dsDNA IgG, IgM, and IgA isotypes and anti-C1q antibody alone or combination in diagnosing SLE. Anti-dsDNA IgG and IgA and anti-C1q were shown to be associated with disease activity, while anti-dsDNA IgM and anti-C1q were associated with lupus nephritis. When the related antibodies were combined, the diagnostic specificity was significantly higher.

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P0492ICAM-1 AND VEGF AS BIOMARKERS FOR PREDICTION LUPUS NEPHRITIS FLARE

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfaa142.p0492 ◽

2020 ◽

Vol 35 (Supplement_3) ◽

Author(s):

Konstantin Chyzh ◽

Tatyana Ryabceva ◽

Nikolay Soroka ◽

Pavel Viarshynin

Keyword(s):

Lupus Nephritis ◽

Blood Serum ◽

Lupus Erythematosus ◽

Roc Analysis ◽

Systemic Disease ◽

Intercellular Adhesion Molecule ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Endothelial Growth Factor ◽

Serum And Urine

Abstract Background and Aims Systemic lupus erythematosus (SLE) is autoimmune systemic disease of the connective tissue and characterized by the development of severe complications associated with damage to vital organs, in particular the kidneys. One of the important issues is an adequate assessment of the current activity and the objective prediction of the lupus nephritis (LN) flare. For this purpose molecular biomarkers are being searched. The work aim is the analyzing role of the intercellular adhesion molecule (ICAM-1) and vascular endothelial growth factor (VEGF) in the serum and urine of patients with SLE as a biomarker for lupus nephritis flare. Method The study group consisted of patients with SLE with LN (n = 23). The control group included patients with SLE without renal pathology (n = 13). The concentration of ICAM-1 and VEGF molecules was determined in blood serum and urine by enzyme-linked immunosorbent assay. Results The median concentration of ICAM-1in serum in patients with LN is 822.41 (580.50; 1300.95) ng/ml. That is two times higher than this indicator in the control group (378.97 (356.47; 439.52) ng/ml). The concentration of the VEGF molecule in serum in the group of LN is 91.99 (74.18; 132.58) ng/ml. That is three times higher than in the control group (31.13 (22.32; 132.58) ng/ml). An increase in the blood serum concentration of patients with SLE ICAM ≥577.7 ng/ml will indicate the presence of LN with a sensitivity of 82.61% and a specificity of 83.33%. ROC analysis showed the AUC coefficient is 0.927 (p = 0.0001). An increase in the blood serum in VEGF concentration of ≥66.43 ng/ml in patients with SLE will indicate the presence of LN with a sensitivity of 80.00% and a specificity of 83.33%. ROC analysis showed that the AUC coefficient is 0.916 (p = 0.006). Conclusion The results allow to conclude that it is possible to use the ICAM and VEGF concentration in serum and urine as early molecular markers of the LN and LN flare in patients with SLE.

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Patients with Proliferative Lupus Nephritis Have Autoantibodies That React to Moesin and Demonstrate Increased Glomerular Moesin Expression

Journal of Clinical Medicine ◽

10.3390/jcm10040793 ◽

2021 ◽

Vol 10 (4) ◽

pp. 793

Author(s):

Dawn J. Caster ◽

Erik A. Korte ◽

Michael L. Merchant ◽

Jon B. Klein ◽

Michelle T. Barati ◽

...

Keyword(s):

Lupus Nephritis ◽

Immune Complex ◽

Lupus Erythematosus ◽

Immunohistochemical Analysis ◽

Enzyme Linked Immunosorbent Assay ◽

Target Antigen ◽

Class Iii ◽

Glomerular Cell ◽

Proliferative Lupus Nephritis ◽

Kidney Involvement

Kidney involvement in systemic lupus erythematosus (SLE)—termed lupus nephritis (LN)—is a severe manifestation of SLE that can lead to end-stage kidney disease (ESKD). LN is characterized by immune complex deposition and inflammation in the glomerulus. We tested the hypothesis that autoantibodies targeting podocyte and glomerular cell proteins contribute to the development of immune complex formation in LN. We used Western blotting with SLE sera from patients with and without LN to identify target antigens in human glomerular and cultured human-derived podocyte membrane proteins. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identified the proteins in the gel regions corresponding to reactive bands observed with sera from LN patients. We identified 102 proteins that were present in both the podocyte and glomerular samples. We identified 10 high-probability candidates, including moesin, using bioinformatic analysis. Confirmation of moesin as a target antigen was conducted using immunohistochemical analysis (IHC) of kidney biopsy tissue and enzyme-linked immunosorbent assay (ELISA) to detect circulating antibodies. By IHC, biopsies from patients with proliferative lupus nephritis (PLN, class III/IV) demonstrated significantly increased glomerular expression of moesin (p < 0.01). By ELISA, patients with proliferative LN demonstrated significantly increased antibodies against moesin (p < 0.01). This suggests that moesin is a target glomerular antigen in lupus nephritis.

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