scholarly journals Establishment of Galectin-3 time-resolved fluoroimmunoassay and its application in idiopathic membranous nephropathy

2021 ◽  
Author(s):  
xiaomei Yu ◽  
Lingli Chen ◽  
Bo Lin ◽  
Li Zhang ◽  
Xue Yang ◽  
...  
Keyword(s):  
Correlation Coefficient ◽  
Membranous Nephropathy ◽  
Idiopathic Membranous Nephropathy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Time Resolved ◽  
Detection Range ◽  
Coefficients Of Variation ◽  
Sandwich Method ◽  
Galectin 3

Abstract Objectives The aim of this study was to establish a time-resolved fluorescent immunoassay (TRFIA) for the detection of serum galactose agglutinin 3 (Gal-3) and apply this method to evaluate the clinical significance of serum Gal-3 in predicting Idiopathic Membranous Nephropathy (IMN) progression. Methods The Gal-3-TRFIA was established using the double antibody sandwich method, with the capture antibodies coated on a 96-well microplate and the detection antibodies chelated to Europium (III) (Eu3+). Serum Gal-3 was detected in 81 patients with IMN and 123 healthy controls to further evaluate the value of the Gal-3 in staging of IMN. Results The sensitivity of the Gal-3-TRFIA assay was 0.85 ng/mL, and the detection range was 0.85–1000 ng/mL. The Gal-3 intra-batch and inter-batch coefficients of variation were 3.45% and 5.12%, respectively. The correlation coefficient (R) between the Gal-3-TRFIA assay and commercially available enzyme-linked immunosorbent assay kits was 0.83. The serum Gal-3 concentration was higher in patients with IMN (65.57 ± 55.90 ng/mL) compared to healthy controls (16.29 ± 9.91 ng/mL, P < 0.0001). Conclusions In this study, a wide detection range Gal-3-TRFIA assay was developed using lanthanide (Eu3+) chelates for the detection of Gal-3 concentrations in serum. The Gal-3-TRFIA also detected the severity of the IMN course.

Anti-Phospholipase A2 Receptor Antibody as Prognostic Indicator in Idiopathic Membranous Nephropathy

2015 ◽  
Vol 42 (3) ◽  
pp. 250-257 ◽  
Author(s):  
Yang Gyun Kim ◽  
Young-Wook Choi ◽  
Se-Yun Kim ◽  
Ju Young Moon ◽  
Chun-Gyoo Ihm ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Membranous Nephropathy ◽  
Remission Rate ◽  
High Titer ◽  
Severe Disease ◽  
Idiopathic Membranous Nephropathy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Receptor Antibody ◽  
Phospholipase A2 Receptor

Background: Anti-phospholipase A2 receptor antibody (PLA2R-Ab) is useful in diagnosing idiopathic membranous nephropathy (IMN). We investigated the clinical relevance of PLA2R-Ab enzyme-linked immunosorbent assay (ELISA) in patients with IMN. Methods: We measured PLA2R-Ab with an ELISA kit from the serum of 160 patients with IMN (n = 93), secondary MN (n = 14) and other glomerulonephritis (n = 41) as well as healthy controls (n = 12) at the time of renal biopsy and investigated the correlation of titers of PLA2R-Ab with clinical parameters. Results: PLA2R-Ab was positive in 41 of 93 patients (44.1%) with IMN. No samples from the patients with secondary MN and other glomerulonephritis or healthy controls were positive with the ELISA test. The PLA2R-Ab-positive patients showed severe disease activity and a low remission rate. The PLA2R-Ab titer positively correlated with proteinuria and was negatively associated with renal function and serum albumin. The patients with a high titer of PLA2R-Ab had significantly decreased remission rates. The cumulative probabilities of remission was significantly lower in patients with PLA2R-Ab (p = 0.01) and even so in patients with a high titer of PLA2R-Ab (p = 0.04). When we compared the ELISA titers with Western blot (WB) data of 43 patients who had been enrolled in our previous study, 18 and 30 patients were positive on ELISA (41.9%) and WB (69.8%), respectively. WB and ELISA had a concordance rate of 72.1% and were positively correlated (r = 0.590, p < 0.001). Conclusion: The presence, as well as a high titer, of PLA2R-Ab on ELISA was associated with poor prognosis of IMN. Assessment of PLA2R-Ab with ELISA is an easy and reliable tool for the diagnosis and guidance of therapeutic plans.

A novel time-resolved fluoroimmunoassay based on magnetic microspheres method for detecting antibodies against Phospholipase A2 Receptor

2021 ◽  
Author(s):  
ting li ◽  
qingqing wu ◽  
Xue Yang ◽  
Yueming liu ◽  
bo lin ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Disease Activity ◽  
Membranous Nephropathy ◽  
Magnetic Microspheres ◽  
Idiopathic Membranous Nephropathy ◽  
Serum Antibodies ◽  
Time Resolved ◽  
Phospholipase A2 Receptor

Background: The level of serum antibodies against the M-type phospholipase A2 receptor (anti-PLA2R-IgG) is closely related to the disease activity of idiopathic membranous nephropathy (IMN). Therefore, the establishment of a...

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.

scholarly journals Clinicopathological features in membranous nephropathy with cancer: A retrospective single-center study and literature review

2019 ◽  
Vol 34 (4) ◽  
pp. 406-413
Author(s):  
Dan Zhang ◽  
Chong Zhang ◽  
Fan Bian ◽  
Wenzhu Zhang ◽  
Gengru Jiang ◽  
...  
Keyword(s):  
Membranous Nephropathy ◽  
Clinicopathological Characteristics ◽  
Idiopathic Membranous Nephropathy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Patients With Cancer ◽  
Phospholipase A2 Receptor ◽  
Single Center Study ◽  
Close Relationship ◽  
Circulating Antibodies

Background: Membranous nephropathy is the most common glomerular disease related to malignancy. However, it is difficult to distinguish between true malignancy-related membranous nephropathy and idiopathic membranous nephropathy coincident with cancer. It has been reported that phospholipase A2 receptor (PLA2R) is the first autoantigen involved in idiopathic membranous nephropathy and thrombospondin type-1 domain-containing 7A (THSD7A) may have a close relationship with malignancy-related membranous nephropathy. Therefore, the aim of this study was to compare the clinicopathological characteristics between membranous nephropathy patients with cancer and idiopathic membranous nephropathy patients without cancer to better detect malignancy-related membranous nephropathy, including glomerular PLA2R and THSD7A depositions and their circulating antibodies, together with glomerular IgG4 deposition. Methods: Twelve membranous nephropathy patients with cancer and 257 idiopathic membranous nephropathy patients without cancer were included in this study and had been followed up for more than 1 year. The glomerular expression of PLA2R, THSD7A, and IgG4 was analyzed by immunohistochemistry. Circulating anti-PLA2R and anti-THSD7A antibodies were assessed by enzyme-linked immunosorbent assay and indirect immunofluorescence testing, respectively. Results: Membranous nephropathy patients with cancer were significantly older and had higher serum creatinine and a lower estimated glomerular filtration rate than idiopathic membranous nephropathy patients ( P<0.05). The positive rates of glomerular PLA2R and IgG4 depositions and circulating anti-PLA2R antibodies in membranous nephropathy patients with cancer were significantly lower than those in idiopathic membranous nephropathy patients without cancer ( P<0.01). Conclusion: The absence of glomerular PLA2R deposition and negative circulating anti-PLA2R antibodies, along with negative glomerular IgG4 staining, may be useful clues to more accurately screen underlying malignancies in membranous nephropathy patients.

scholarly journals Potential role of soluble TNF-α receptors in diagnosis of idiopathic membranous nephropathy

2018 ◽  
Vol 16 ◽  
pp. 205873921877608
Author(s):  
Chunhua Xun ◽  
Yong Zhao ◽  
Wenjuan Wang ◽  
Tiantian Cheng
Keyword(s):  
Tumor Necrosis Factor ◽  
Healthy Volunteers ◽  
Sensitivity And Specificity ◽  
Membranous Nephropathy ◽  
Tumor Necrosis ◽  
Idiopathic Membranous Nephropathy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Urea ◽  
Tnf Α ◽  
Necrosis Factor

Soluble tumor necrosis factor alpha (TNF-α) receptors contain two receptors: soluble tumor necrosis factor receptor (sTNFR) 1 and 2, and the aim of our study was to discover their concentration and diagnostic value for idiopathic membranous nephropathy (IMN). In total, 58 patients with IMN, 51 patients with chronic kidney disease (CKD), and 30 healthy volunteers were enrolled in this study. Levels of serum sTNFR1 and sTNFR2 were determined by enzyme-linked immunosorbent assay (ELISA). Serum cystatin C (CysC), urea, creatinine (CREA), uric acid (UA), total protein (TP), albumin (ALB), and 24-h urinary protein (proteinuria, PRO) were examined by automatic biochemical analyzer. Levels of sTNFR1 and sTNFR2 were significantly higher in IMN group than CKD and control group ( P < 0.05). In IMN group, there were significant correlation between sTNFR1 and sTNFR2 ( P < 0.01). Both sTNFR1 and sTNFR2 were positively related to serum urea, CREA, CysC, UA, 24-h PRO ( P < 0.05) and negatively related to ALB ( P < 0.01). Receiver operating characteristic (ROC) analysis showed that the area under the curve (AUC) of sTNFR1 and sTNFR2 were 0.997 and 0.993, respectively, when control was healthy volunteers. When sTNFR1 cut-off was 959.15 pg/mL, the sensitivity and specificity were 96.6% and 100%, respectively. When sTNFR2 cut-off was 2449.43 pg/mL, the sensitivity and specificity were 93.1% and 100%, respectively. While the control was CKD group, AUC of sTNFR1 and sTNFR2 were 0.647 and 0.626, respectively. When sTNFR1 cut-off was 3356.57 pg/mL, the sensitivity and specificity were 72.4% and 60.8%, respectively. When sTNFR2 cut-off was 6497.34 pg/mL, the sensitivity and specificity were 72.4% and 58.8%, respectively. This is the first study to show that both levels of sTNFR1 and sTNFR2 increased and correlated with serum urea, CREA, CysC, UA, ALB, 24-h PRO and could be usable for IMN diagnosis and differential diagnosis between IMN and CKD.

scholarly journals Diagnostic accuracy of anti-phospholipase A2 receptor (PLA2R) antibodies in idiopathic membranous nephropathy: an Italian experience

2020 ◽  
Author(s):  
Brunetta Porcelli ◽  
Andrea Guarnieri ◽  
Fabio Ferretti ◽  
Guido Garosi ◽  
Lucia Terzuoli ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Clinical Diagnosis ◽  
Membranous Nephropathy ◽  
Immunosuppressive Treatment ◽  
Positive Likelihood Ratio ◽  
Idiopathic Membranous Nephropathy ◽  
Renal Diseases ◽  
Enzyme Linked Immunosorbent Assay ◽  
Special Focus ◽  
Phospholipase A2 Receptor

Abstract Background Autoantibodies against-phospholipase A2 receptor (PLA2R) are specific markers of idiopathic membranous nephropathy (iMN). Enzyme-linked immunosorbent assay (ELISA) is becoming the preferred method in many laboratories for the determination of anti-PLA2R antibodies, because it provides quantitative results, and is not prone to subjective interpretation, as is the case with indirect immunofluorescence assay. Methods The purpose of our study was to determine the diagnostic performance of serum PLA2R antibodies detected by commercially available ELISA in a large Italian multicenter cohort of patients with biopsy-proven iMN and in patients with other renal diseases, with special focus on evaluating the optimal cut-off value to discriminate positive and negative results. A total of 495 consecutive patients were recruited. Renal biopsies were performed in all patients, and blood samples were taken before the initiation of immunosuppressive treatment. Results According to the clinical diagnosis and to kidney biopsy, 126 patients were diagnosed with iMN and 369 had other non-membranous nephropathies. Anti-PLA2R autoantibodies were detected using a commercial anti-PLA2R ELISA. At a cut-off value of 20 relative units (RU)/ml indicated by the manufacturer for positive classification, sensitivity was 61.1% and specificity 99.7%. At a cut-off value of 14 RU/ml indicated by the manufacturer for borderline results, sensitivity was 63.5% and specificity remained the same (99.7%). At a cut-off of 2.7 RU/ml, selected as the optimal cut-off on the basis of ROC curve analysis, sensitivity was 83.3% and specificity 95.1%. The best overall efficiency of the test was observed at 2.7 RU/ml; however, the highest positive likelihood ratio and diagnostic odds ratio were achieved at 14 RU/ml. A cut-off threshold higher than 14 RU/ml or lower than 2.7 RU/ml entailed worse test performance. Conclusion Depending on the clinical use (early diagnosis or as a support to confirm clinical diagnosis), nephrologists may take advantage of this evidence by choosing the most convenient cut-off. However, renal biopsy remains mandatory for the definitive diagnosis of iMN and for the assessment of disease severity.

scholarly journals A PLA2R-IgG4 Antibody-Based Predictive Model for Assessing Risk Stratification of Idiopathic Membranous Nephropathy

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Xiaobin Liu ◽  
Jing Xue ◽  
Xiaoyi Guo ◽  
Yijie Ding ◽  
Yi Zhang ◽  
...  
Keyword(s):  
Random Forest ◽  
Risk Stratification ◽  
Membranous Nephropathy ◽  
Predictive Ability ◽  
Risk Groups ◽  
Idiopathic Membranous Nephropathy ◽  
Quantitative Detection ◽  
Time Resolved ◽  
Phospholipase A2 Receptor ◽  
The Impact

Background. Known as an autoimmune glomerular disease, idiopathic membranous nephropathy (IMN) is considered to be associated with phospholipase A2 receptor (PLA2R) in terms of the main pathogenesis. The quantitative detection of serum PLA2R-IgG and PLA2R-IgG4 antibodies by time-resolved fluoroimmunoassay (TRFIA) was determined, and the value of them, both in the clinical prediction of risk stratification in IMN, was observed in this study. Methods. 95 patients with IMN proved by renal biopsy were enrolled, who had tested positive for serum PLA2R antibodies by ELISA, and the quantitative detection of serum PLA2R-IgG and PLA2R-IgG4 antibodies was achieved by TRFIA. All the patients were divided into low-, medium-, and high-risk groups, respectively, which were set as dependent variables, according to proteinuria and renal function. Random forest (RF) was used to estimate the value of serum PLA2R-IgG and PLA2R-IgG4 in predicting the risk stratification of progression in IMN. Results. Out-of-bag estimates of variable importance in RF were employed to evaluate the impact of each input variable on the final classification accuracy. The variable of albumin, PLA2R-IgG, and PLA2R-IgG4 had high values (>0.3) of 0.3156, 0.3981, and 0.7682, respectively, which meant that these three were more important for the risk stratification of progression in IMN. In order to further assess the contribution of PLA2R-IgG and PLA2R-IgG4 to the model, we built four different models and found that PLA2R-IgG4 played an important role in improving the predictive ability of the model. Conclusions. In this study, we established a random forest model to evaluate the value of serum PLA2R-IgG4 antibodies in predicting risk stratification of IMN. Compared with PLA2R-IgG, PLA2R-IgG4 is a more efficient biomarker in predicting the risk of progression in IMN.

scholarly journals Circulating CD14+CD163+CD206+ M2 Monocytes Are Increased in Patients with Early Stage of Idiopathic Membranous Nephropathy

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Jie Hou ◽  
Manli Zhang ◽  
Yuhong Ding ◽  
Xinrui Wang ◽  
Tao Li ◽  
...  
Keyword(s):  
Membranous Nephropathy ◽  
Early Stage ◽  
Clinical Signs ◽  
Idiopathic Membranous Nephropathy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Monocyte Subset ◽  
Blood Monocytes ◽  
Cell Counts ◽  
Potential Association ◽  
Phospholipase A2 Receptor

Aim. To analyze changes in peripheral blood monocytes and their clinical significance in patients with early stage of idiopathic membranous nephropathy (IMN). Methods. A total of 27 patients with early stage of IMN and 16 age- and sex-matched healthy controls (HCs) were recruited for the study. The monocyte subset counts in circulation were measured by flow cytometry, and serum interleukin- (IL-) 10 and IL-12 concentrations were tested by enzyme-linked immunosorbent assay. The potential association between clinical signs and monocyte subset counts was analyzed statistically. Results. Compared with the HCs, the patients with early stage of IMN had higher counts of CD14+CD163+, CD14+CD163+CD206+, and CD14+CD163+CD206+CD115+ M2-like monocytes. The CD14+CD163+CD206+ M2-like cell counts and intracellular IL-10 concentrations in the monocytes were positively correlated with progression in proteinuria. The levels of serum IL-10 were significantly higher in early IMN patients than in the HCs. Furthermore, CD14+CD163+CD206+ M2-like cell counts in the patients with incipient IMN were also positively related with 24 h urinary albumin levels and the values of serum M-type phospholipase A2 receptor (PLA2R). Conclusion. CD14+CD163+CD206+ M2-like monocytes may contribute to the pathologic process in early-stage IMN and could serve as potential markers for evaluating the disease severity.

scholarly journals Urinary Matrix Metalloproteinase-9 and Nephrin in Idiopathic Membranous Nephropathy: A Cross-Sectional Study

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Akankwasa Gilbert ◽  
An Changjuan ◽  
Cheng Guixue ◽  
Liu Jianhua ◽  
Qin Xiaosong
Keyword(s):  
Matrix Metalloproteinase ◽  
Membranous Nephropathy ◽  
Healthy Subjects ◽  
Kidney Injury ◽  
Matrix Metalloproteinase 9 ◽  
Idiopathic Membranous Nephropathy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cross Sectional ◽  
Glomerular Diseases ◽  
Metalloproteinase 9

Aim. Idiopathic membranous nephropathy (IMN) has a varied clinical course that requires accurate prediction as a prerequisite for treatment administration. Currently, its prognosis relies on proteinuria, a clinical parameter whose onset lags behind kidney injury. Increased urinary excretion of matrix metalloproteinase-9 (MMP-9) and nephrin has been reported in a number of IMN-like glomerular diseases in which they reflected disease severity. However, little or nothing is known of the importance of these biomarkers in IMN, a major cause of adult nephrotic syndrome. To highlight their potential, we measured both biomarkers and assessed their relationships with key parameters of renal function in IMN. Methods. We quantified urinary MMP-9 and nephrin in 107 biopsy-proven IMN patients and 70 healthy subjects by enzyme-linked immunosorbent assay (ELISA). We then compared biomarker levels between patients and healthy subjects and among patients with different clinical features. We also determined the relationship of each biomarker with proteinuria and the estimated glomerular filtration rate (eGFR). Results. Urinary MMP-9 and nephrin were significantly higher in IMN compared to healthy controls. Unlike nephrin, MMP-9 correlated significantly with proteinuria and was significantly higher among patients with nephrotic range proteinuria. Both biomarkers were correlated with eGFR, but only MMP-9 was significantly higher in patients with eGFR less than 90 ml/min/1.73 m2. Conclusion. Our findings suggest that urinary MMP-9 holds a greater potential than urinary nephrin in monitoring the severity of IMN.

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