Quantitative analysis of type IV collagen subchains in the glomerular basement membrane of patients with Alport syndrome with confocal microscopy

Abstract Context.—Alport syndrome and thin glomerular basement membrane nephropathy (TBMN) are genetically heterogenous conditions characterized by structural abnormalities in the glomerular basement membrane and an initial presentation that usually involves hematuria. Approximately 40% of patients with TBMN are heterozygous carriers for autosomal recessive Alport syndrome, with mutations at the genetic locus encoding type IV collagen α3 [α3(IV)] and α4 chains. However, although the clinical course of TBMN is usually benign, Alport syndrome, particularly the X-linked form with mutations in the locus encoding the α5 chain of type IV collagen [α5(IV)], typically results in end-stage renal disease. Electron microscopy is essential to diagnosis of TBMN and Alport syndrome on renal biopsy, although electron microscopy alone is of limited value in distinguishing between TBMN, the heterozygous carrier state of X-linked Alport syndrome, autosomal recessive Alport syndrome, and even early stages of X-linked Alport syndrome. Objectives.—To review diagnostic pathologic features of each of the above conditions, emphasizing the need for immunohistology for α3(IV) and α5(IV) in addition to electron microscopy to resolve this differential diagnosis on a renal biopsy. The diagnostic value of immunofluorescence studies for α5(IV) on a skin biopsy in family members of patients with Alport syndrome also is reviewed. Data Sources.—Original and comprehensive review articles on the diagnosis of Alport syndrome and TBMN from the past 35 years, primarily the past 2 decades, and experience in our own renal pathology laboratory. Conclusions.—Although Alport syndrome variants and TBMN do not show characteristic light microscopic findings and can be difficult to differentiate from each other even by electron microscopy, using a combination of electron microscopy and immunohistology for α3(IV) and α5(IV) enables pathologists to definitively diagnose these disorders on renal biopsy in most cases.

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Spectrum of collagen type IV nephropathies: From thin basement membrane nephropathy to Alport syndrome

Srpski arhiv za celokupno lekarstvo ◽

10.2298/sarh08s4323v ◽

2008 ◽

Vol 136 (Suppl. 4) ◽

pp. 323-326 ◽

Cited By ~ 1

Author(s):

Alenka Vizjak ◽

Dusan Ferluga

Keyword(s):

Basement Membrane ◽

Glomerular Basement Membrane ◽

Alport Syndrome ◽

Type Iv Collagen ◽

Mutation Screening ◽

Glomerular Sclerosis ◽

Collagen Type Iv ◽

Dominant Type ◽

Thin Basement Membrane Nephropathy ◽

Type Iv

Alport syndrome and thin basement membrane nephropathy are common causes of persistent familial haematuria. They are associated with various mutations in type IV collagen genes. Mutations in genes, coding for ?5 chain of collagen IV, cause X-linked Alport syndrome, whereas mutations in genes for ?3 and ?4 chains can cause the autosomal recessive and autosomal dominant type of Alport syndrome or benign familial haematuria with thin basement membrane nephropathy. In view of the wide spectrum of phenotypes, an exact diagnosis is sometimes difficult to achieve. Few studies of genotype-phenotype correlations in Alport syndrome have shown that various types of mutations may be a significant predictor of the severity of disease. Histopathologic findings in Alport syndrome vary from normal kidney to nonspecific focal segmental and global glomerular sclerosis with characteristic ultrastructural finding of thickening and splitting of the glomerular basement membrane. Thin basement membrane nephropathy is characterized by diffuse thinning of the glomerular basement membrane on an ultrastructural level, while by light microscopy glomeruli are mostly unremarkable. Because of present limitations of mutation screening techniques, kidney biopsy with mandatory ultrastructural analysis and immunohistochemistry examination for type IV collagen ? chains remains a standard approach for establishing diagnosis and determining the mode of transmission of the disease.

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Mutation in alpha 5(IV) collagen chain gene in nonfamilial hematuria.

Journal of the American Society of Nephrology ◽

10.1681/asn.v62264 ◽

1995 ◽

Vol 6 (2) ◽

pp. 264-268

Author(s):

K Kitagawa ◽

K Nakanishi ◽

K Iijima ◽

H Nishio ◽

Y Sado ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Basement Membrane ◽

Glomerular Basement Membrane ◽

Alport Syndrome ◽

Type Iv Collagen ◽

Chain Reaction ◽

Col4a5 Gene ◽

Abnormal Expression ◽

Type Iv ◽

Polymerase Chain

Alport syndrome is an inherited disorder characterized by progressive nephritis with ultrastructural basket-weave changes of the glomerular basement membrane and neurosensory deafness. Mutations in the COL4A5 gene encoding the Type IV collagen alpha 5 chain have been reported to occur in patients with X-linked Alport syndrome. A girl with hematuric nephritis, characteristic basket-weave glomerular basement membrane changes, and abnormal expression of the Type IV collagen alpha 5 chain immunohistochemically, but no family history of nephritis, was identified. Mutation detection enhancement gel electrophoresis of the polymerase chain reaction-amplified exons of COL4A5 from this patient revealed a sequence variant in the exon 50 region. Sequence analysis of her polymerase chain reaction product demonstrated a single-base (C; nucleotide 4728 from the 5' end) deletion in exon 50. This novel mutation alters the reading frame and introduces a translation stop codon that would be expected to result in a noncollagenous domain with only 209, instead of the normal 229, amino acid residues. Gene tracking with restriction enzyme AfIIII demonstrated that her mother was normal. These findings represent a new mutation of the X-linked Alport syndrome in this patient and demonstrate that a COL4A5 gene mutation causes the abnormal expression of Type IV collagen alpha 5 chain protein.

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IDENTIFICATION OF ??3, ??4, AND ??5 CHAINS OF TYPE IV COLLAGEN AS ALLOANTIGENS FOR ALPORT POSTTRANSPLANT ANTI-GLOMERULAR BASEMENT MEMBRANE ANTIBODIES

Transplantation Journal ◽

10.1097/00007890-200002270-00038 ◽

2000 ◽

Vol 69 (4) ◽

pp. 679-684 ◽

Cited By ~ 42

Author(s):

Raghu Kalluri ◽

Adriana Torre ◽

Charles F. Shield ◽

Eric D. Zamborsky ◽

Michelle C. Werner ◽

...

Keyword(s):

Basement Membrane ◽

Glomerular Basement Membrane ◽

Type Iv Collagen ◽

Type Iv

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Glomerular Basement Membrane Selective Permeability in Short-term Streptozotocin-induced Diabetic Rats

International Journal of Experimental Diabetes Research ◽

10.1155/edr.2000.19 ◽

2000 ◽

Vol 1 (1) ◽

pp. 19-30 ◽

Cited By ~ 10

Author(s):

Michele Doucet ◽

Irene Londoño ◽

Amparo Gómez-Pascual ◽

Moise Bendayan

Keyword(s):

Basement Membrane ◽

Glomerular Basement Membrane ◽

Type Iv Collagen ◽

Serum Proteins ◽

Diabetic Rats ◽

Glomerular Permeability ◽

Structural Alterations ◽

Early Stages ◽

Type Iv ◽

Basement Membrane Thickening

In diabetes, the glomerular basement membrane undergoes thickening and structural alterations with loss of glomerular permselectivity properties. However, the onset of the alterations at early phases of diabetes is unclear. Aiming to determine the functional and structural alterations of the glomerular wall in the early stages of diabetes, we have studied the distribution of endogenous circulating albumin and type IV collagen in the glomerular basement membrane, using the immunocytochemical approach. The streptozotocin-injected hyperglycemic rat was our animal model. Renal tissues were examined after 10 days, 2, 4 and 6 months of hyperglycemia. Upon immunogold labelings, changes in the glomerular permeability to endogenous albumin were found altered as early as upon ten days of hyperglycemia. In contrast, no structural modifications were detected at this time point. Indeed, glomerular basement membrane thickening and an altered type IV collagen labeling distribution were only observed after four months of hyperglycemia, suggesting that functional alterations take place early in diabetes prior to any structural modification. In order to evaluate the reversibility of the glomerular alterations, two-month-old diabetic animals were treated with insulin. These animals showed a significant restoring of their glomerular permselectivity. Our results suggest a link between glycemic levels and alteration of glomerular permeability in early stages of diabetes, probably through high levels of glycated serum proteins.

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Glomerular basement membrane synthesis and serum concentration of type IV collagen in streptozotocin-diabetic rats

Diabetologia ◽

10.1007/bf00281124 ◽

1984 ◽

Vol 26 (2) ◽

Cited By ~ 44

Author(s):

Ch. Hasslacher ◽

R. Reichenbacher ◽

F. Gechter ◽

R. Timpl

Keyword(s):

Basement Membrane ◽

Serum Concentration ◽

Glomerular Basement Membrane ◽

Type Iv Collagen ◽

Diabetic Rats ◽

Membrane Synthesis ◽

Type Iv ◽

Basement Membrane Synthesis ◽

Streptozotocin Diabetic Rats

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RETRACTED: Production and Characterization of Recombinant Rat Non-collagen Domain of α3 Chain of Type IV Collagen α3 (IV) NC1 Antigen

European Scientific Journal ESJ ◽

10.19044/esj.2016.v12n24p98 ◽

2016 ◽

Vol 12 (24) ◽

pp. 98

Author(s):

Afsana Munni

Keyword(s):

Basement Membrane ◽

Glomerular Basement Membrane ◽

Type Iv Collagen ◽

Kidney Diseases ◽

Cell Epitope ◽

Affinity Column ◽

Igg Antibody ◽

Amino Terminal ◽

Type Iv ◽

Collagenous Domain

Glomerulonephritis disease is characterized by inflammation of glomeruli or small blood vessels in the kidney which causes kidney diseases. Glomerulonephritis disease deposits the anti-GBM auto antibody in the glomerular basement membrane. The type IV collagen is the main component of glomerular basement membrane that has α3 chain of type (IV) collagen of non-collagenous domain which contains N-terminal 7S domain, a triple helical collagenous domain, and a C- terminal non-collagenous glomerular domain (NC1). The amino terminal of α3 (IV) NC1 that induces the experimental autoimmuno glomerulonephritis (EAG) in rat model has been identified. The recombinant rat α3 (IV)NC1 antigen has nine amino acid span that is consistent with antibody or T cell epitope which is induced in EAG. The research is carried out on the recombinant rat α3 (IV) NC1 production, purification, quantification, and characterization. The circulation of Anti-GBM antibody in glomerular basement membrane can be measured by the ELISA assay. In addition, the recombinant rat antigen is secreted in HEK293 cell supernatant which is purified by Anti-FLAG M2 monoclonal IgG antibody affinity column. In addition, it is characterized and quantified by SDS-PAGE gel electrophoresis and Western blotting techniques.

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The prevalence and immunological features of anti-glomerular basement membrane antibody in patients with HIV

10.21203/rs.2.21447/v2 ◽

2020 ◽

Author(s):

Wenjing Wang ◽

Xiao-yu Jia Jia ◽

Zhao Cui ◽

Yan Chen ◽

Wei Wang ◽

...

Keyword(s):

Basement Membrane ◽

Glomerular Basement Membrane ◽

Type Iv Collagen ◽

Solid Phase ◽

Chimeric Protein ◽

Kidney Injury ◽

Enzyme Linked Immunosorbent Assay ◽

Hiv Viral Load ◽

Hiv Patients ◽

Type Iv

Abstract Background: Anti-glomerular basement membrane disease (GBM) is a kind of chronic autoimmune disease caused by the deposition of circulating anti-GBM antibodies. Non-collagen region of α3 chain of type IV collagen (α3(IV)NC1) is one of the main target antigens. And on α3, EA and EB are the most classical antigen epitopes. It has been reported that anti-GBM antibodies can be detected in HIV patients, but its immunological characteristics are still unclear. In this study, the immunological characteristics of the target antigens were clarified. Methods: Total 93 HIV patients and 20 healthy volunteers were selected in Beijing Youan Hospital from 2017 to 2018. Recombinant human α1-α5(IV)NC1, chimeric protein EA and EB were used as solid phase antigens. Enzyme-linked immunosorbent assay was employed to measure concentrations and subtypes of serum IgG autoantibodies specifically against GBM.Results: Five out of the 93 patients with HIV had low to moderate levels of anti-GBM antibodies. However, these patients presented with no clinical manifestation of any kidney injury or pulmonary hemorrhages. Compared with HIV patients with negative antibodies, there were no significant differences in gender, age, CD4+T cell count and HIV viral load. All sera of five patients recognized non-collagenous domain1 (NC1) of alpha 3 chain of type IV collagen [(α3(IV)NC1] as classic anti-GBM patients, followed by α5(IV)NC1. The antibodies against α3(IV)NC1 were IgG3 predominant, but did not react with either of the classic epitopes on α3 (EA and EB).Conclusion: These data suggest a distinct immunological profile of anti-GBM antibodies in patients with HIV, and might explain the non-pathogenic features of HIV associated anti-GBM antibodies.

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