scholarly journals Proteomic Analysis of Glomerular Proteins in Primary and Bucillamine-induced Membranous Nephropathy

2021 ◽  
Author(s):  
Hajime Kaga ◽  
Hirotoshi Matsumura ◽  
Takehiro Suzuki ◽  
Naoshi Dohmae ◽  
Masafumi Odaka ◽  
...  
Keyword(s):  
Membranous Nephropathy ◽  
Proteomic Analysis ◽  
Kidney Diseases ◽  
Principal Component ◽  
Control Group ◽  
Drug Induced ◽  
Protein Levels ◽  
Disease Marker ◽  
Phospholipase A2 Receptor ◽  
Associated Proteins

Abstract The aim of this study was to characterize glomerular proteins in primary membranous nephropathy (pMN) and drug-induced secondary MN (sMN) by laser microdissection and comparative proteomic analysis. We used renal biopsy specimens from 6 patients with anti-phospholipase A2 receptor autoantibody (PLA2R Ab) (+) pMN, 6 patients with PLA2R Ab (‒) pMN, 6 patients with bucillamine (BCL)-induced sMN, and 5 control cases (time 0 transplant biopsies). Proteins were extracted from laser-microdissected glomeruli and analyzed using mass spectrometry. The quantification values of protein abundance in each MN group were compared with those in the control group. More than 800 proteins with high confidence were identified. Principal component analysis revealed a different distribution between the pMN and sMN groups. For further analysis, 441 proteins matched with ≥3 peptides were selected. Among the pMN and sMN groups, we compared the profiles of several protein groups based on the structural and functional characteristics, such as immunoglobulins, complements, complement-regulating proteins, podocyte-associated proteins, glomerular basement membrane proteins, and several proteins that are known to be associated with kidney diseases, including MN. Between the pMN and BCL-induced sMN groups, we observed common and different alterations in protein levels such as known disease-associated proteins and potential disease marker proteins.

MO432COMPARATIVE ANALYSIS OF INDIRECT IMMUNOFLUORESCENCE AND ENZYME-LINKED IMMUNOSORBENT ASSAYS FOR ANTI-PLA2R ASSESMENT IN PATIENTS WITH PRIMARY MEMBRANOUS NEPHROPATHY

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Olga Galkina ◽  
Evdokia Bogdanova ◽  
Irina Zubina ◽  
Elena Levykina ◽  
Alexei Smirnov
Keyword(s):  
Indirect Immunofluorescence ◽  
Membranous Nephropathy ◽  
Laboratory Diagnosis ◽  
Response To Therapy ◽  
Control Group ◽  
Course Of Disease ◽  
Phospholipase A2 Receptor ◽  
Gender And Age ◽  
Positive Results

Abstract Background and Aims Antibodies to M-type phospholipase A2 receptor (PLA2R-Ab) are considered to be a promising biomarker for laboratory diagnosis of primary membranous nephropathy (PMN) and may be useful in the evaluation of the response to therapy and CKD prognosis. The aim of the study was to compare two immunoassay methods – indirect immunofluorescence (IIF) and enzyme immunoassay (ELISA) for the determination of circulating PLA2R-Ab in patients with PMN. Method The study included 54 patients aged 55 (40-63) yrs. (M: F [33:21]) with PMN before treatment (n=16) and treated with immunosuppressive therapy (IST) (n=38), and apparently healthy individuals of the corresponding gender and age (n=10). Proteinuria and estimated glomerular filtration rate (eGFR) were determined in all participants. The levels of PLA2R-Ab were determined by IIF and quantitative/ semi-quantitative ELISA (EURUIMMUN AG test, Germany). In 16 PMN patients without treatment and 28 PMN patients treated with IST the level of PLA2R-Ab was measured one time and in 10 PMN patients treated IST – in dynamic, from 2 to 5 times. Statistical comparisons among groups were performed using Mann–Whitney U-test and Kruskal-Wallis H tests. The association between variables was estimated using Spearman’s coefficient. Sensitivity and specificity of the methods were calculated. Results The correlation coefficient between IIF and ELISA was 0.82 (p <0.005). There were more PLA2R-Ab-positive cases detected by ELISA, both before treatment (ELISA - 80%, IIF - 67%) and among patients treated with IST (ELISA - 63%, IIF - 50%). In control group, ELISA showed no positive results for PLA2R-Ab (specificity was 100%). The levels of proteinuria and eGFR were associated with autoantibodies determined by ELISA, both quantitative and semi-quantitative (proteinuria: r = 0.69, p = 0.001; eGFR: r = -0.38, p = 0.035) but not by IIF (proteinuria: r=0.33, p=0.061; eGFR: r=-0.26, p=0.082). The levels of PLA2R-Ab measured by ELISA correlated with the course of disease in patients treated with IST, while IIF did not show any dynamics is some cases. Conclusion Both quantitative and semi-quantitative ELISA were considered to be more preferable methods since the obtained results correlate with renal dysfunction and allow to assess the concentration of PLA2R-Ab in the course of disease more accurately, that may contribute to timely correction of treatment and improvement of outcome.

scholarly journals Phospholipase A2 Receptor Antibody IgG4 Subclass Improves Sensitivity and Specificity in the Diagnosis of Idiopathic Membranous Nephropathy

2019 ◽  
Vol 44 (4) ◽  
pp. 848-857 ◽  
Author(s):  
Biao Huang ◽  
Yi Zhang ◽  
Liang Wang ◽  
Wenwei Xu ◽  
Jue Zhang ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Sensitivity And Specificity ◽  
Membranous Nephropathy ◽  
Kidney Diseases ◽  
Measurement Range ◽  
Idiopathic Membranous Nephropathy ◽  
Time Resolved ◽  
Specificity And Sensitivity ◽  
Phospholipase A2 Receptor ◽  
Highly Sensitive

Aims: The aim of this study was to develop a new method for detecting anti-phospholipase A2 receptor-IgG4 to improve the sensitivity and specificity in the diagnosis of idiopathic membranous nephropathy (IMN). Methods: A highly sensitive quantitative assay was developed for the detection of serum anti-phospholipase A2 receptor-IgG4 with europium chelation by time-resolved fluoroimmunoassay (TRFIA), and a mouse anti-human IgG4 tracer was prepared using europium chelation for detection. The specificity and sensitivity of anti-phospholipase A2 receptor-IgG4 in the diagnosis of IMN were further assessed in patients with different kidney diseases. Results: The detection limit of anti-PLA2R-IgG4 was 0.69 ng/mL. The measurement range of anti-PLA2R-IgG4 TRFIA was 0.69–2,500 ng/mL. Mean serum anti-PLA2R-IgG4 was 21.27 ± 15.15 ng/mL in 45 healthy volunteers, 31.08 ± 18.17 ng/mL in 29 IgA nephropathy patients, 49.10 ± 34.32 ng/mL in 8 lupus nephropathy patients, and 10,324.11 ± 17,030.40 ng/mL in 30 IMN patients. The anti-PLA2R-IgG4 cutoff concentration was >161.2 ng/mL with the sensitivity of 90.0% and specificity of 100% in the diagnosis of IMN. However, the cutoff for other kidney diseases was lower than 161.2 ng/mL. Conclusion: The serum anti-phospholipase A2 receptor IgG4 detected with the method developed in this study has higher sensitivity and higher specificity than total IgG in the diagnosis of IMN.

Conjugated Linoleic Acid Causes a Marked Increase in Liver α-Tocopherol and Liver α-Tocopherol Transfer Protein in C57BL/6 J Mice

2010 ◽  
Vol 80 (1) ◽  
pp. 65-73 ◽  
Author(s):  
Pei-Min Chao ◽  
Wan-Hsuan Chen ◽  
Chun-Huei Liao ◽  
Huey-Mei Shaw
Keyword(s):  
Antioxidant Enzymes ◽  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Thiobarbituric Acid ◽  
Control Group ◽  
Thiobarbituric Acid Reactive Substances ◽  
Control Groups ◽  
Protein Levels ◽  
Transfer Protein ◽  
And Control

Conjugated linoleic acid (CLA) is a collective term for the positional and geometric isomers of a conjugated diene of linoleic acid (C18:2, n-6). The aims of the present study were to evaluate whether levels of hepatic α-tocopherol, α-tocopherol transfer protein (α-TTP), and antioxidant enzymes in mice were affected by a CLA-supplemented diet. C57BL/6 J mice were divided into the CLA and control groups, which were fed, respectively, a 5 % fat diet with or without 1 g/100 g of CLA (1:1 mixture of cis-9, trans-11 and trans-10, cis-12) for four weeks. α-Tocopherol levels in plasma and liver were significantly higher in the CLA group than in the control group. Liver α-TTP levels were also significantly increased in the CLA group, the α-TTP/β-actin ratio being 2.5-fold higher than that in control mice (p<0.01). Thiobarbituric acid-reactive substances were significantly decreased in the CLA group (p<0.01). There were no significant differences between the two groups in levels of three antioxidant enzymes (superoxide dismutase, glutathione peroxidase, and catalase). The accumulation of liver α-tocopherol seen with the CLA diet can be attributed to the antioxidant potential of CLA and the ability of α-TTP induction. The lack of changes in antioxidant enzyme protein levels and the reduced lipid peroxidation in the liver of CLA mice are due to α-tocopherol accumulation.

Multifactorial analysis of clinical laboratory signs, the levels of IL-17A, IL-23, IL-33, IL-35, and specific antibodies in the serum of patients with Lyme borreliosis without erythema migrans

2020 ◽  
Vol 4 (11) ◽  
pp. 676-681
Author(s):  
V.V. Sapozhnikova ◽  
◽  
A.L. Bondarenko ◽  
Keyword(s):  
Principal Component Analysis ◽  
Lyme Borreliosis ◽  
Clinical Laboratory ◽  
Acute Infection ◽  
Erythema Migrans ◽  
Principal Component ◽  
Component Analysis ◽  
Control Group ◽  
Multifactorial Analysis ◽  
Specific Antibodies

Aim: to determine the association between clinical laboratory parameters, the production of cytokines (IL-17A, -23, -33, -35), and specific IgM and IgG in the serum of patients with Lyme borreliosis without erythema migrans. Patients and Methods: complete blood count, the concentrations of IL-17A, -23, -33, -35, and the levels of specific IgM and IgG were measured during acute infection and convalescence (n=30). The control group included age- and sex-matched healthy individuals (n=30). Statistical analysis was performed using the StatSoft Statistica v 10.0 software (parametric and non-parametric methods and multifactorial analysis, i.e., principal component analysis). Results: most (80%) patients with Lyme borreliosis without erythema migrans are the people of working age. In most patients, the combination of the specific antibodies against Borrelia afzelii and Borrelia garinii (76.7%) and severe intoxication and inflammatory process (100%) were detected. Moderate and severe disease associated with meningism was diagnosed in 90% and 10%, respectively. The mean duration of hectic period was 8.3±1.27 days. Abnormal ECG was reported in 40% of patients, i.e., conduction abnormalities in 20%, sinus bradycardia in 16.7%,and sinus tachycardia in 3.3%. The clinical laboratory signs of hepatitis without jaundice were identified in 26.7%. During treatment, the significant reduction in band and segmented neutrophil counts as well as the significant increase in platelet count were revealed compared to these parameters at admission. Abnormal cytokine levels (i.e., the increase in IL-17A, -23, -33 and the deficiency of IL-35) were detected. Conclusions: multifactorial analysis has demonstrated that the severity of immunological abnormalities in patients with Lyme borreliosis without erythema migrans is associated with fever, cardiac and liver disorders, the high levels of IL-23 and IL-33, and the lack of IL-35 and specific IgM and IgG. KEYWORDS: tick-borne borreliosis, Lyme disease without erythema migrans, clinical laboratory signs, cytokines, specific antibodies, multifactorial analysis, principal component analysis. FOR CITATION: Sapozhnikova V.V., Bondarenko A.L. Multifactorial analysis of clinical laboratory signs, the levels of IL-17A, IL-23, IL-33, IL-35, and specific antibodies in the serum of patients with Lyme borreliosis without erythema migrans. Russian Medical Inquiry. 2020;4(11):676–681. DOI: 10.32364/2587-6821-2020-4-11-676-681.

Case Review of lgG4-Related Disease Patient with Combined Tubulointerstitial Nephritis and Membranous Nephropathy Coexisting with Cholangiocarcinoma: Case Report

2020 ◽  
Vol 103 (11) ◽  
pp. 1230-1235
Keyword(s):  
Renal Failure ◽  
Membranous Nephropathy ◽  
Tubulointerstitial Nephritis ◽  
Disease Patient ◽  
Case Review ◽  
Related Disease ◽  
Phospholipase A2 Receptor ◽  
Wide Range ◽  
Serum Igg4 ◽  
Secondary Membranous Nephropathy

Immunoglobulin G4-related disease (IgG4-RD) has recently been recognized as an autoimmune disorder involving multiple organs. The kidney is a represented organ with a wide range of renal manifestations. The authors report a case of an 83-year-old Thai male with combined IgG4 tubulointerstitial nephritis and membranous nephropathy coexisting with cholangiocarcinoma. The patient presented with proteinuria, acute renal failure, eosinophilia, hypocomplementemia, and high serum IgG4 concentration. The diagnosis was IgG 4-related tubulointerstitial nephritis and membranous nephropathy on renal biopsy, with negative immunohistochemistry for anti-phospholipase A2 receptor antibodies. Magnetic resonance imaging (MRI) abdomen showed two wedge shaped arterial enhancing lesions of liver. Liver biopsy revealed adenocarcinoma, compatible with cholangiocarcinoma. Proteinuria and renal failure were resolved with initial steroid treatment. Meanwhile, IgG4-related membranous nephropathy should be considered in the differential diagnosis for patients with proteinuria. Potentially, IgG4-RD may be rarely associated with carcinoma development. However, further studies are recommended to ratify and confirm the association between IgG4-RD and incidence of malignancies. Keywords: IgG4-related disease, Membranous nephropathy, Secondary membranous nephropathy, Tubulointerstitial nephritis, Cholangiocarcinoma

scholarly journals Autism Spectrum Disorder (ASD) with and without Mental Regression is Associated with Changes in the Fecal Microbiota

Nutrients ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 337 ◽  
Author(s):  
Julio Plaza-Díaz ◽  
Antonio Gómez-Fernández ◽  
Natalia Chueca ◽  
María Torre-Aguilar ◽  
Ángel Gil ◽  
...  
Keyword(s):  
Intestinal Microbiota ◽  
Present Report ◽  
Principal Component ◽  
Autism Spectrum ◽  
Control Group ◽  
Healthy Group ◽  
Sequencing Platform ◽  
Preliminary Results ◽  
Children With Asd ◽  
Mental Regression

New microbiome sequencing technologies provide novel information about the potential interactions among intestinal microorganisms and the host in some neuropathologies as autism spectrum disorders (ASD). The microbiota–gut–brain axis is an emerging aspect in the generation of autistic behaviors; evidence from animal models suggests that intestinal microbial shifts may produce changes fitting the clinical picture of autism. The aim of the present study was to evaluate the fecal metagenomic profiles in children with ASD and compare them with healthy participants. This comparison allows us to ascertain how mental regression (an important variable in ASD) could influence the intestinal microbiota profile. For this reason, a subclassification in children with ASD by mental regression (AMR) and no mental regression (ANMR) phenotype was performed. The present report was a descriptive observational study. Forty-eight children aged 2–6 years with ASD were included: 30 with ANMR and 18 with AMR. In addition, a control group of 57 normally developing children was selected and matched to the ASD group by sex and age. Fecal samples were analyzed with a metagenomic approach using a next-generation sequencing platform. Several differences between children with ASD, compared with the healthy group, were detected. Namely, Actinobacteria and Proteobacteria at phylum level, as well as, Actinobacteria, Bacilli, Erysipelotrichi, and Gammaproteobacteria at class level were found at higher proportions in children with ASD. Additionally, Proteobacteria levels showed to be augmented exclusively in AMR children. Preliminary results, using a principal component analysis, showed differential patterns in children with ASD, ANMR and AMR, compared to healthy group, both for intestinal microbiota and food patterns. In this study, we report, higher levels of Actinobacteria, Proteobacteria and Bacilli, aside from Erysipelotrichi, and Gammaproteobacteria in children with ASD compared to healthy group. Furthermore, AMR children exhibited higher levels of Proteobacteria. Further analysis using these preliminary results and mixing metagenomic and other “omic” technologies are needed in larger cohorts of children with ASD to confirm these intestinal microbiota changes.

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