Expression Of Phospholipase A2 Receptor Research Articles

A Clinicopathological Analysis of Membranous Nephropathy and Its Correlation With the Immunohistochemical Expression of Phospholipase A2 Receptor (PLA2R) in Renal Biopsies in a Tertiary Care Center

A Clinicopathological Analysis of Membranous Nephropathy and Its Correlation With the Immunohistochemical Expression of Phospholipase A2 Receptor (PLA2R) in Renal Biopsies in a Tertiary Care Center

Distribution spectrum and clinical significance of glomerular exostosin (EXT1) deposits in PLA2R-positive membranous nephropathy.

The discovery of antigen phospholipase A2 receptor (PLA2R) in 2009 ushered in the antigen-based study of membranous nephropathy. Thefurther putative antigen exostosin 1/2 (EXT1/2) was described in 2019. However, the distribution spectrum of glomerular EXT1 deposits in membranous nephropathy has not been fullyelucidated. We conducted a retrospective cohort study of biopsy-proven membranous nephropathy patients. Patients with complete baseline data and adequate tissue specimens were included in this study. Tests for glomerular expression of PLA2R and EXT1 and circulating anti-PLA2R antibodies were performed. Clinicopathological and outcome data were reviewed. We included 626 patients, namely, 487 (77.8%) PLA2R-positive patients and 54 (8.6%) EXT1-positive patients; 32 (5.1%) patients were dual-positive for PLA2R and EXT1 (PLA2R + /EXT1 +). A higher percentage of dual-positive patients had low C3 levels (P < 0.001) and were more likely to have autoimmune diseases (P = 0.013) than PLA2R-positive and EXT1-negative (PLA2R + /EXT1-) patients. Kidney biopsy findings revealed that there was a higher percentage of glomerular IgG1, IgG2, IgA, C4, and C1q deposits (P < 0.05), "full-house" staining (P < 0.001), and stronger intensity of C1q staining (P = 0.002) in PLA2R + /EXT1 + patients. Based on Kaplan-Meier analysis, a higher percentage of PLA2R + /EXT1 + patients exhibited partial or complete remission of proteinuria. Furthermore, EXT1-positive expression was a favourable predictor for proteinuria remission, whereas interstitial fibrosis/tubular atrophy was an unfavourable predictor. A complement C3 level < 0.79g/L was independently associated with EXT1 positivity in PLA2R-positive membranous nephropathy. We describe a subgroup of PLA2R and EXT1 dual-positive patients. Patients in this subset exhibited more signs of autoimmunity and morefrequent clinical remission. In PLA2R-positive membranous nephropathy, a complement C3 level < 0.79g/L was independently associated with EXT1 positivity, which was a favourable predictor for proteinuria remission.

Comprehensively characterizing cellular changes and the expression of THSD7A and PLA2R1 under multiple in vitro models of podocyte injury.

The unique morphology and gene expression of podocytes are critical for kidney function, and their abnormalities lead to nephropathies such as diabetic nephropathy and membranous nephropathy. Podocytes cultured in vitro are valuable tools to dissect the molecular mechanism of podocyte injury relative to nephropathy, however, these models have never been comprehensively compared. Here, we comprehensively compared the morphology, cytoskeleton, cell adhesion, cell spreading, cell migration, and lipid metabolism under five commonly used in vitro models including lipopolysaccharide (LPS), puromycin aminonucleoside (PAN), doxorubicin (Dox), high glucose, and glucose deprivation. Our results indicate that all stimulations significantly downregulate the expression of synaptopodin both in human and mouse podocytes. All stimulations affect podocyte morphology but show different intensity and phenotypes. In general, thefive stimulations reduce cell adhesion, cell spreading, and cell migration, but the effect in human and mouse podocytes is slightly different. Human podocytes show high expression of genes enriched in the pentose phosphate pathway. Dox and PAN treatment show a strong effect on gene expression in lipid metabolism, while the other three stimulations show minimal effect. The expression of phospholipase A2 receptor (PLA2R1) and type-1 domain-containing protein 7 A (THSD7A) show opposite trends in given cells. Stimulations can dramatically affect the expression of PLA2R1 and THSD7A. Inhibition of super-enhancers reduces PLA2R1 and THSD7A expression, but ERK inhibition enhances their expression. Our results demonstrate distinctive responses in five commonly used in vitro podocyte injury models and the dynamic expression of PLA2R1 and THSD7A, which supply novel information to select suitable podocyte injury models.

Neural Epidermal Growth Factor-Like 1 Protein-Positive Membranous Nephropathy in Chinese Patients.

The neural EGF-like 1 (NELL-1) protein is a novel antigen in primary membranous nephropathy. The prevalence and clinical characteristics of NELL-1-positive membranous nephropathy in Chinese individuals with primary membranous nephropathy are unclear. A total of 832 consecutive patients with biopsy-proven primary membranous nephropathy were enrolled. The glomerular expression of phospholipase A2 receptor (PLA2R) and thrombospondin type 1 domain-containing 7A (THSD7A) was screened. Glomerular immunohistochemistry staining for NELL-1 was performed in 43 patients with PLA2R- and THSD7A-negative membranous nephropathy, 31 patients with PLA2R-positive membranous nephropathy, and two patients with PLA2R and THSD7A double positivity. The NELL-1 antibody was also detected in the sera of patients with NELL-1-positive membranous nephropathy by western blot. Clinical and pathologic features were comparable between patients with isolated NELL-1-positive, isolated PLA2R/THSD7A-positive, and triple antigen-negative membranous nephropathy. Among the 832 patients with primary membranous nephropathy, 11 of 54 (20%) patients with PLA2R-negative membranous nephropathy had THSD7A-positive membranous nephropathy. NELL-1-positive membranous nephropathy accounted for 35% (15 of 43) of all patients with PLA2R- and THSD7A-negative membranous nephropathy. One patient was double positive for NELL-1 and PLA2R in glomerular deposits and positive for only the PLA2R antibody in the serum. Most patients with NELL-1-positive membranous nephropathy were women. No tumors were found. There were significant differences in the prevalence of IgG subtypes between patients with different antigen positivity. Among patients with isolated NELL-1-positive membranous nephropathy, although 80% (12 of 15) were IgG4 staining positive, the proportion of IgG4 dominance was only 67% (ten of 15). About one third of patients who were PLA2R and THSD7A negative were NELL-1 positive in Chinese patients with primary membranous nephropathy. NELL-1-positive membranous nephropathy was more common than THSD7A-positive membranous nephropathy in PLA2R-negative membranous nephropathy.

Effects of phospholipase A2 receptor and thrombospondin type-1 domain-containing 7A expression in glomerular basement membranes on treatment response and renal outcome in membranous nephropathy.

The aim of this study was to define the clinicopathologic features of phospholipase A2 receptor (PLA2R) and/or thrombospondin type-1 domain-containing 7A (THSD7A) associated membranous nephropathy(MN) focusing on their impact to disease relapse and response to treatment. A total of 201 patients were enrolled for baseline clinical and histopathological features and 102 patients with a clinical follow-up for more than 1year were evaluated for outcomes. Immunohistochemical staining was performed with PLA2R and THSD7A antibodies on kidney biopsies and glomerular staining was evaluated. PLA2R expression was observed in 75% of the patients' biopsies; however, THSD7A expression was present only in 7 patients' biopsies (3.5%). No significant difference was found between histopathological and clinical features of PLA2R positive and negative patients, collectively. Glomerular PLA2R expression was significantly associated with complete and complete/partial remission with first-line treatment; however, overall complete, and complete/partial remission rates did not differ from PLA2R negative patients (p = 0.2 and p = 0.8). Male gender, the presence of IgG4 staining and a necessity of immunosuppressive treatment were significantly associated with glomerular PLA2R expression. One patient, who developed end-stage renal disease, had glomerular expression for both PLA2R and THSD7A. Three patients with THSD7A-positive MN achieved complete remission. The probability of achieving complete remission is high in patients with PLA2R-positive MN for whom the relapse rate was also higher. The overall renal outcome did not differ from PLA2R negative cases. Low incidence of THSD7A-positive MN reduces the possibility of future randomized controlled trials.

Renal expression of PLA2R, THSD7A, and IgG4 in patients with membranous nephropathy and correlation with clinical findings.

Membranous nephropathy (MN) is the most common cause of nephrotic syndrome in nondiabetic adults. M-type phospholipase A2 receptor (PLA2R), thrombospondin type-1 domain-containing 7A (THSD7A) are known as target podocyte antigens in membranous nephropathy (MN). Antibodies against these podocyte antigens are used in the initiation of treatment and response monitoring. However, the relationship between renal podocyte antigens and treatment response is not clear yet. We evaluated the relationship between immunohistochemical PLA2R, THSD7A and IgG4 staining, clinical findings and treatment response in kidney biopsies. Fifty-eight patients with MN were included in this retrospective study. In the renal biopsy samples of the patients, PLA2R, THSD7A and IgG4 were stained immunohistochemically and evaluated by light microscopy. The clinical, laboratory and treatment results of the patients were obtained from the hospital records. The study included a total of 58 patients with MN and a mean follow-up period of 32.3±19.7months. In patients with primary MN; PLA2R, THSD7A and IgG4 were positive in 57.1% (n=28), 12.2% (n=6) and 69.4% (n=34), respectively. Only PLA2R staining was distinctly higher in patients with primary MN than secondary MN (P=.025). Dual positivity (PLA2R+THSD7A) was detected in five (10.2%) of patients with primary MN. We did not determine any relationship between the PLA2R, THSD7A and IgG4 staining patterns and treatment response of the patients. It was found no correlation between PLA2R, THSD7A and IgG4 staining in kidney tissue and treatment response. Interestingly, dual positivity (PLA2R+THSD7A) was detected only in primary MN.

Tacrolimus Prevents TWEAK-Induced PLA2R Expression in Cultured Human Podocytes.

Primary membranous nephropathy is usually caused by antibodies against the podocyte antigen membrane M-type phospholipase A2 receptor (PLA2R). The treatment of membranous nephropathy is not fully satisfactory. The calcineurin inhibitor tacrolimus is used to treat membranous nephropathy, but recurrence upon drug withdrawal is common. TNF superfamily members are key mediators of kidney injury. We have now identified key TNF receptor superfamily members in podocytes and explored the regulation of PLA2R expression and the impact of tacrolimus. Data mining of single cell transcriptomics and glomerular transcriptomics data identified TNFRSF12a/Fn14 as the highest expressed TNF receptor superfamily gene in human membranous nephropathy, and this was confirmed by immunohistochemistry that also identified NFκB activation in membranous nephropathy podocytes. Additionally, glomerular transcriptomics identified PLA2R1 expression as being increased in membranous nephropathy in the parenteral administration of the Fn14 ligand TWEAK increased podocyte PLA2R expression in mice. Furthermore, in cultured human podocytes, TWEAK increased the expression of PLA2R as well as the expression of other genes recently identified by GWAS as linked to membranous nephropathy: NFKB1 and IRF4. Interestingly, IRF4 encodes the FK506-binding protein 52 (FKBP52), a protein associated with tacrolimus. Tacrolimus prevented the increased expression of PLA2R, NFKB1 and IRF4 induced by TWEAK in cultured podocytes. In conclusion, TWEAK upregulates the expression of PLA2R and of other genes linked to membranous nephropathy in podocytes, and this is prevented by tacrolimus. An impact of tacrolimus on the expression of PLA2R and other genes in podocytes may underlie its efficacy in treating the disease as well as the frequent recurrence of nephrotic syndrome upon tacrolimus withdrawal.

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

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. 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. 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). 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.

SUN-030 PHOSPHOLIPASE A2 RECEPTOR AND THROMBOSPONDIN TYPE-1 DOMAIN-CONTAINING 7A IN MALIGNANCY MEMBRANOUS NEPHROPATHY

Phospholipase A2 receptor (PLA2R) and thrombospondin type-1 domain-containing 7A (THSD7A) have recently been identified as pathogenic antigens in patients with membranous nephropathy (MN). Notably, PLA2R is detected in a few patients with malignancy-associated MN. Furthermore, a high incidence of cancer is reported in patients with THSD7A-associated MN. It is thus necessary to clarify the usefulness of PLA2R and THSD7A in malignancy-associated MN. Serum anti-PLA2R antibodies and glomerular PLA2R expression were first screened in 36 patients with malignancy-associated MN by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence, respectively, followed by examination of serum anti-THSD7A antibodies and glomerular THSD7A. THSD7A expression in cancer tissues was also assessed in 9 of the 36 patients. Among the 36 patients, 12 (33%) were positive for both glomerular PLA2R staining and serum anti-PLA2R antibodies, and 2 were positive for either glomerular PLA2R staining or serum anti-PLA2R antibodies. IgG4 was the dominant IgG subclass in PLA2R-related patients. One patient with positve glomerular PLA2R staining was also positive for THSD7A staining. Among the remaining 22 (61%) patients who were double negative for glomerular PLA2R staining and serum anti-PLA2R antibodies, 17 of 20 (85%) had IgG1 dominant deposits in glomeruli, and 2 (9%) were positive for glomerular THSD7A staining. Serum anti-THSD7A antibody was not detected in any of the 36 patients. Among the 9 patients with available cancer tissues, enhanced expression of THSD7A in the cancer tissues was observed for 5 (56%), and one also showed enhanced glomerular expression of THSD7A. Screening of glomerular PLA2R antigen and serum anti-PLA2R antibodies is necessary in patients with malignancy-associated MN, whereas the incidence of glomerular THSD7A antigen or circulating anti-THSD7A antibodies is uncommon.

Compared staining of the phospholipase A2 receptor in the glomeruli of Chinese adults and children with idiopathic membranous nephropathy

BackgroundThe identification of the M-type phospholipase A2 receptor (PLA2R) is a breakthrough recognized as a major target for adults with idiopathic membranous nephropathy (IMN). However, the role PLA2R played in pediatric patients with IMN, particularly in Chinese, has yet to be determined. MethodsThis retrospective study included 187 adult patients and 38 pediatric patients aged 17 years or younger with biopsy proved IMN. The pediatric cohort consisted of 27 children aged from 1 to 12 years and 11 children aged from 13 to 17. Glomerular expression of PLA2R was analyzed in stored, formalin-fixed, paraffin-embedded kidney biopsy sections. ResultsPLA2R staining in glomerular deposits was observed in 82.7% and 42.1% of adult and pediatric patients with IMN, respectively. The PLA2R-positive staining patients with IMN presented with more severe clinical features than PLA2R-negative staining patients in both adult and pediatric cohorts. When compared to the young children patients with IMN, the adolescents exhibited a higher positive rate of PLA2R staining (81.8% versus 25.9%), similar to the adult patients. ConclusionThe clinical features and prevalence of PLA2R positive staining in adolescent patients with IMN were similar to adult patients, suggesting that they probably have a close etiology and pathogenesis. However, most of the young children patients with IMN were PLA2R negative staining, suggesting a different underlying etiology.

MiR-130a-5p prevents angiotensin II-induced podocyte apoptosis by modulating M-type phospholipase A2 receptor

ABSTRACT Podocyte apoptosis is considered as the important element that promotes the development and progress of membranous nephropathy (MN). Unfortunately, the underlying mechanism of podocytes apoptosis in MN remains elusive. We compared the renal expressions of miR-130a-5p and M-type phospholipase A2 receptor (PLA2R) between MN patients (n = 30) and 30 controls by qRT-PCR and western blot, respectively. The podocyte damage model in vitro was established by angiotensin II (Ang II, 100 nmol/L) exposure for 24 h. Interaction between miR-130a-5p and PLA2R was determined using dual-luciferase reporter gene assay. MN mice were induced by intravenous injection of cBSA. In this study, miR-130a-5p expression was significantly decreased both in the renal biopsy specimens from MN patients and podocyte cell line AB8/13 following stimulation of Ang II. Overexpressed miR-130a-5p in AB8/13 cells significantly attenuated the Ang II induced-apoptosis in vitro. In contrast, down-regulated miR-130a-5p induced podocyte apoptosis. PLA2R was identified as the target of miR-130a-5p in AB8/13 cells. And up-regulated or down-regulated PLA2R could obviously attenuate the effect of miR-130a-5p overexpression or knockdown on the apoptosis of AB8/13 cells. Furthermore, it was also observed that overexpressed miR-130a-5p by miR-130a-5p agomir could obviously alleviate renal injury in MN mice. In conclusion, decreased miR-130a-5p was contributed to the pathological mechanism of MN through increasing PLA2R expression, which induced podocyte apoptosis.

Anti-Phospholipase A2 Receptor Antibodies as Diagnostic Modality in Primary Idiopathic Membranous Nephropathy

Background Phospholipase A2 receptor (PLA2R), an IgG4 subclass antibody, is being extensively studied for diagnosis, monitoring and prognostication of cases of primary membranous nephropathy (MN). There is significant heterogeneity within the studies with most of the work targeted towards circulating PLA2R antibody. The present study evaluates PLA2R glomerular staining in primary MN as a diagnostic test in conjunction with the serology. Methods: Diagnostic study. We studied paraffin-embedded kidney biopsies of 60 patients with biopsy-proven MN, for the presence of PLA2R in glomerular immune deposits utilising immunohistochemistry. 60 cases of non MN cases along with autopsy renal tissue were used as controls. A subset of patients (n=40) were tested for circulating PLA2R antibody using quantitative sandwich enzyme immunoassay. Results: Positive PLA2R expression was present in 37/39 (94.8%) cases of primary MN, 5/21 (23.8% ) of secondary MN and 4/60 (6.6%) of non MN group. The expression was detected along glomerular basement membrane in granular pattern of varying intensities. Circulating anti-PLA2R antibodies were detected in 71.42% of primary MN cases, 8.3% of secondary MN cases and non MN group. PLA2R immunostaining exhibited higher sensitivity and modest specificity for differentiating primary from secondary MN (ROC-AUC :0.885 of IHC vs ROC-AUC: 0.776 of serology). The immunostaining however required precision and expertise to read the results. Conclusion: Tissue staining of PLA2R antibody in renal biopsies will be complementary to serology and two done in conjunction will be better discriminator between primary and secondary MN.

Clinical and Histological Features of Phospholipase A2 Receptor-Associated and Thrombospondin Type-I Domain-containing 7A-Associated Idiopathic Membranous Nephropathy: A Single Center Retrospective Study from China

BackgroundM-type phospholipase A2 receptor (PLA2R) was identified as the major target antigen in idiopathic membranous nephropathy (IMN). Another target antigen, namely thrombospondin type-1 domain-containing 7A (THSD7A), was recently detected in approximately 10% of non-PLA2R-associated IMN. In this single center retrospective study, clinical and histological features of PLA2R-associated and THSD7A-associated IMN patients were evaluated.Material/MethodsA total of 192 IMN patients, who were receiving no glucocorticoids or immunosuppressant before renal biopsy, were enrolled in this study and followed for a median duration of 25.5 months. IMN with enhanced glomerular PLA2R and THSD7A staining by immunohistochemistry (IHC) were designated as PLA2R-associated IMN and THSD7A-associated IMN respectively. Serum anti-PLA2R and anti-THSD7A antibodies levels were assessed by enzyme linked immunosorbent assay and indirect immunofluorescence testing in PLA2R-associated and THSD7A-associated IMN.ResultsOf 192 IMN patients, 164 patients (85.4%) had PLA2R-associated IMN and 3 patients (1.6%) had THSD7A-associated IMN. Compared with non-PLA2R-associated IMN patients, the 24-hour urinary protein levels were significantly higher (P=0.008), whereas, the proteinuria remission rates were significantly lower (P=0.03) in PLA2R-associated IMN patients. No pathological differences were found between PLA2R-associated IMN and non-PLA2R-associated IMN. Among 3 THSD7A-associated IMN patients, 1 patient had elevated serum anti-THSD7A antibody levels, which was below detectable levels after achieving partial proteinuria remission with combined glucocorticoid and cyclosporine treatment.ConclusionsCompared with non-PLA2R-associated IMN patients in our cohort, PLA2R-associated IMN patients presented with more severe proteinuria and lower remission rates after treatment, with no distinct histological differences. Glomerular expression of PLA2R could be a useful marker to indicate the severity, treatment response, and prognosis of IMN.

The features of idiopathic membranous nephropathy in elderly patients and its correlation with the expression of the expression of phospholipase A2 receptor

Objective To investigate the clinico-pathological features of idiopathic membranous nephropathy(IMN)and the expression of phospholipase A2 receptor(PLA2R)in elderly patients. Methods A total of 109 elderly patients with IMN confirmed by renal biopsy at Wuxi People′s Hospital from July 2008 to February 2015 were included.Data were retrospectively collected. Results (1)Participating patients with IMN had a mean age of(67.3±5.4)years, and 67.9% of them had hypertension and 65.1% had nephrotic syndrome.Compared with non-elderly patients, elderly patients had a higher proportion with hypertension(67.9% vs.25.2%)(P=0.000), higher systolic pressure[(143.1±15.2)mmHg vs.(127.3±13.3)mmHg](P=0.000), higher diastolic pressure[(88.4±10.0)mmHg vs.(80.2±8.4)mmHg](P=0.000), more severe tubulointerstitial lesions[(3.1±1.9) points vs.(2.0±1.9)points](P=0.000), and lower eGFR[(70.9±22.9)ml·min-1·1.73 m-2vs.(90.6±27.1)ml·min-1·1.73 m-2](P=0.000). (2)There were more severe tubulointerstitial lesions[(4.7±1.8)points vs.(2.4±1.7)points, 2.9±1.6 points](P=0.000, 0.000)and lower eGFR[(50.4±17.4)ml·min-1·1.73 m-2vs.(80.3±19.7)ml·min-1·1.73 m-2, (72.3±21.4)ml·min-1·1.73 m-2](P=0.000, 0.000)in elderly patients of pathological stage Ⅱ, compared with patients of pathological stages Ⅰ and Ⅰ-Ⅱ.(3)The rate of positive PLA2R was 82.4%.Patients with positive PLA2R had higher proteinuria[(4.5±2.3)g vs.(2.9±1.1)g](P=0.042), lower eGFR[(66.8±21.8)ml·min-1·1.73 m-2vs.(97.7±16.0)ml·min-1·1.73 m-2](P=0.000), and more severe tubulointerstitial lesions[(3.1±2.0)points vs.(1.7±1.1)points](P=0.037)than patients with negative PLA2R.(4)Multiple regression analysis showed that PLA2R positive rate(P=0.008), tubulointerstitial lesion(P=0.000), and level of cholesterol(P=0.025)were negatively correlated with eGFR(R2=0.572). Conclusions Compared with non-elderly patients, elderly patients with IMN have poorer prognosis as a result of higher blood pressure and more severe tubulointerstitial lesions.Elderly patients with IMN of advanced pathological stages and positive PLA2R have more severe kidney injury and tubulointerstitial lesions, resulting in poor prognosis. Key words: Nephropathy, membranous; Pathology, clinical; Receptors, phospholipase A2

Expression of phospholipase A2 receptor and IgG4 in patients with membranous nephropathy.

ObjectivesThe aims of this study were to detect the expression of M phospholipase A2 receptor (PLA2R) in the kidney tissue of patients with idiopathic membranous nephropathy (IMN), secondary membranous nephropathy (SMN), and the nonmembranous nephropathy (non-MN), to evaluate the value of PLA2R in the kidney tissue and serum anti-PLA2R antibody in the diagnosis of membranous nephropathy (MN), and to explore the relationship between PLA2R of the kidney tissue or serum anti-PLA2R antibody and clinical features of MN.MethodsThe kidney tissue was collected by kidney biopsy. Immunofluorescence assay was used to detect the level of PLA2R and IgG4 antigen in kidney tissue. Furthermore, the level of the PLA2R was detected using the enzyme-linked immunosorbent assay (ELISA). The positive and negative rates of PLA2R and IgG4 in different diseases and the sensitivity and specificity, were calculated using the statistical method. The specificity and coincidence rate of PLA2R or anti-PLA2R used in the differential diagnosis of IMN and SMN were evaluated.ResultsThe expression intensities of anti-PLA2R antibody and IgG4 were significantly higher in patients with IMN than in patients with SMN but are not non-MN. There was no significant difference in anti-PLA2R antibody and IgG4 in patients with SMN and non-MN. The coincidence rate of serum anti-PLA2R antibody and PLA2R in kidney tissue was 100%.ConclusionThe expression of PLA2R and IgG4 antibody had great significance in the pathological diagnosis of MN. The detection of the serum anti-PLA2R antibody had great diagnostic value in diagnosing MN.

Anti-phospholipase A2 receptor antibody positive hepatitis B virus-associated membranous nephropathy remitted with entecavir after relapse with lamivudine

Background: Phospholipase A2 receptor (PLA2R) is thought to be an intrinsic antigen of idiopathic membranous nephropathy (IMN), and has been widely used for the differentiation from secondary membranous nephropathies. However, the positive expression of PLA2R in the patients with hepatitis B virus associated membranous nephropathy (HBV-MN) is controversial in Asian countries, because co-localization of PLA2R and HBV antigens in glomeruli have been reported. Case Presentation: We report a case of anti PLA2R antibody positive HBV-MN that was remitted with entecavir after a relapse during treatment with lamivudine. In a renal biopsy of the case, we could confirm the co-localized glomerular deposition of HBV surface antigen (HBs-Ag) and PLA2R using double staining of immunofluorescence. We also could observe the relapse of nephrotic syndrome correlated with the increased titer of HBs-Ag, and the remission with the decrease of HBs-Ag by the change of antiviral agents. Conclusions: Our case demonstrated that the renal manifestation of HBV-MN clearly responded to antiviral agents. Furthermore, the co-localized glomerular depositions of PLA2R and HBs-Ag in HBV-MN may be concerned with the etiology of MN patients with chronic HBV infection.

Circulating Antibodies against Thrombospondin Type-I Domain-Containing 7A in Chinese Patients with Idiopathic Membranous Nephropathy.

Thrombospondin type-I domain-containing 7A (THSD7A) was recently identified as the target antigen in about 10% of patients with M-type phospholipase A2 receptor (PLA2R)-negative membranous nephropathy in European and North American populations. The prevalence of THSD7A in other populations and their clinical associations deserve further clarification. Immunofluorescence assay was performed to investigate anti-THSD7A antibodies in 578 consecutive patients with biopsy-proven idiopathic membranous nephropathy, 114 patients with secondary membranous nephropathy, 64 disease controls, and 20 healthy controls. Glomerular expression of THSD7A antigen was examined by immunohistochemistry. Anti-PLA2R antibodies and glomerular PLA2R expression were also screened. Among the 578 patients with idiopathic membranous nephropathy, 12 (2%) patients were identified as THSD7A-positive: ten patients were THSD7A-positive alone, which accounted for 16% (ten of 64) of PLA2R-negative patients; two patients were dual-positive for both anti-THSD7A and anti-PLA2R antibodies and showed enhanced expression of both antigens colocalized in glomeruli. Among the 114 patients with secondary membranous nephropathy, one among 44 (2%) patients with cancer had anti-THSD7A antibodies, whereas 18 of 44 (41%) had anti-PLA2R antibodies. No anti-THSD7A antibody was detected in other disease controls or healthy individuals. Clinical features were comparable between the patients with and without THSD7A. During follow-up, two patients who achieved remission had a clearance of circulating antibodies against THSD7A, whereas antibodies increased in parallel with proteinuria in a patient with a relapse. THSD7A-associated membranous nephropathy has a low prevalence in Chinese patients. The double-positive patients suggest dual autoimmune responses.

Diagnostic value of renal phospholipase A2 receptor and serum anti-phospholipase A2 receptor antibody in membranous nephropathy

To examine the expression of phospholipase A2 receptor (PLA2R) in renal tissues and the level of anti-PLA2R antibody in serum in patients with idiopathic membranous nephropathy (IMN) and secondary membranous nephropathy (SMN), and to evaluate their diagnostic value in IMN. Methods: A total of 73 patients, who were diagnosed between May, 2014 and February, 2015 in the Department of Nephrology of the Second Xiangya Hospital, Central South University, were divided into three groups: an IMN group (n=48), an SMN group (n=17) and a minimal change disease group (n=8) according to the renal biopsy. PLA2R expression in renal tissues and the level of anti-PLA2R antibody in serum were detected by indirect immunofluorescence technique. Results: The positive rate and fluorescence intensity for PLA2R in the renal tissues in the IMN group were higher than those in the SMN group (91.7% in the IMN group vs 29.4% in the SMN group, P<0.05), while the positive rate and serum level for anti-PLA2R antibody in the IMN group were higher than those in the SMN group (85.4% in the IMN group vs 29.4% in the SMN group, P<0.05); the expression of PLA2R in renal tissues and the serum level for anti-PLA2R antibody were not detected in the minimal change disease group. The serum level of anti-PLA2R antibody was positively correlated with 24 h urine protein (r=0.432, P<0.01) and negatively correlated with serum albumin (r=-0.307, P<0.05). Conclusion: The expression of PLA2R in renal tissues and the serum level of anti-PLA2R antibody might be potential markers for diagnosis of IMN.

Renal Phospholipase A2 Receptor and the Clinical Features of Idiopathic Membranous Nephropathy.

Background:According to the renal phospholipase A2 receptor (PLA2R) immunohistochemistry, idiopathic membranous nephropathy (iMN) could be categorized into PLA2R-associated and non-PLA2R-associated iMN. This study aimed to examine whether the non-PLA2R-associated iMN had any difference in clinical features compared with PLA2R-associated iMN.Methods:A total of 231 adult patients diagnosed as iMN were recruited to this retrospective study. Renal PLA2R expression was examined by immunofluorescence. Among these patients, 186 (80.5%) with complete baseline clinical data were used for further study. Urinary protein excretion, serum albumin, and creatinine were analyzed. For those patients with follow-up longer than 1 year, the relationship between PLA2R and response to immunosuppressants were analyzed. The t-test was used for parametric analysis and the Mann-Whitney U-test was used for nonparametric analysis. Categorical variables were described as frequencies or percentages, and the data were analyzed with Pearson's Chi-square test or Fisher's exact test.Results:Of the 231 iMN patients, 189 showed renal detectable PLA2R expression (81.8%). The baseline serum creatinine, serum albumin, and urine protein excretion were not significantly different between PLA2R-associated (n = 145) and non-PLA2R-associated iMN patients (n = 41). However, about 1/3 of the non-PLA2R-associated iMN had abnormal serological tests, significantly more common than PLA2R-associated iMN (31.7% vs. 8.3%, P = 0.000). The non-PLA2R-associated iMN had lower C4 levels compared with PLA2R-associated iMN (P = 0.004). The non-PLA2R-associated iMN patients also showed a better response to immunosuppressants (complete remission [CR] 42.9%; partial remission [PR] 14.3%) compared with PLA2R-associated iMN (CR 3.2%; PR 48.4%, P = 0.004) at the 3rd month.Conclusions:There were no significant differences in serum creatinine, albumin, and urine protein excretion between PLA2R-associated and non-PLA2R-associated iMN, while the non-PLA2R-associated iMN patients showed more abnormal serological tests. The non-PLA2R-associated iMN seemed to respond more quickly to the immunosuppressive therapy compared with PLA2R-associated iMN.

Immunohistochemical Glomerular Expression of Phospholipase A2 Receptor in Primary and Secondary Membranous Nephropathy: A Retrospective Study in an Indian Cohort with Clinicopathological Correlations

Background: Limited published literature exists on the utility and standardization of anti-phospholipase A2 receptor (anti-PLA2R) immunohistochemistry (IHC) for the diagnosis of primary membranous nephropathy (MN). The study aimed to validate anti-PLA2R IHC for the diagnosis of primary MN and clinicopathological correlations in an Indian cohort. Methods: Subjects included patients with primary and secondary MN diagnosed between January 2012 and August 2014 with an adequate renal biopsy and at least 1 year of clinical follow-up. Anti-PLA2R IHC was performed in all cases with miscellaneous renal lesions as controls. Electron microscopy was performed in selected cases. Sensitivity and specificity of anti-PLA2R IHC to identify primary MN was evaluated. Histopathological analyses of primary and secondary MN were done with clinicopathological correlations including serum creatinine, eGFR, chronic kidney disease stage, 24-h urine protein, serum cholesterol, serum albumin, and hypertension at presentation and follow-up, using the Kruskal-Wallis test and Spearman rank correlation. A p value of ≤0.05 was considered statistically significant. Results: In 153 MN patients (99 primary, 54 secondary) and 37 miscellaneous controls, anti-PLA2R IHC differentiated primary from secondary MN with a sensitivity of 70.2% and a specificity of 96.6%. Secondary MN had increased mesangial matrix expansion compared to primary MN (p = 0.001). Severe nephrotic syndrome, impaired renal function, and hypertension were all more common in primary than in secondary MN. Conclusion: Anti-PLA2R IHC is a specific marker to distinguish primary MN from secondary MN.