Abstract
The glomerulonephritis disease is characterized by inflammation of glomeruli or small blood vessels in the kidney that causes kidney diseases. The reason of glomerulonephritis disease is to deposit 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 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 spans that are consistent with antibody or T cell epitope that induces 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 that is purified by Anti-FLAG M2 monoclonal IgG antibody affinity column and characterized and quantified by SDS-PAGE gel electrophoresis and Western blotting techniques.
Highlights
This type IV collagen was transfected to the embryonic kidney (HEK) 293 cells (Invitrogen, San Diego, CA) using cell supernatant [27]
The cells line secreted the recombinant rat α3 (IV) NC1 that were cultured by adding Dulbeccos Modified Eagle Medium (DMEM), fetal bovine serum, penicillin streptomycin solution
Glomerulonephritis disease consists of Goodpasture’s syndrome with pulmonary hemorrhage causes of circulating and tissue bonding antibodies to the glomerular basement membrane The majority of antibodies activity is in the α3 (IV) NC1 domain [30]-[32] and antigen of NC1 domain can bind to the Anti-GBM autoantibody in basement membrane
Summary
The kidneys have renal artery, nephron, glomerulus, tubule, capillaries, collecting duct parts. The nephron consists of glomerulus, tubules, capillaries, and collecting duct. The filtered blood exits from glomerulus and enters the tubules and collecting ducts. The autoimmune diseases can be defined as the abnormal function of body immune system that produces antibodies (T-cell and B-cell autoreactivity) against normal host cells and tissues. Glomerulonephritis is the direct cause of capillary wall damage and disturbances in normal glomerular function, gradually loss the filtration capacity and excessive transfer for hematuria and proteinuria. Depositioning of antibodies within the glomeruli involved the immune system of glomerulonephritis It can be explained by antibody mediated mechanism either by direct reaction glomerular antigens (Anti-GBM disease) or by deposition of antibodies in glomeruli. The non-proliferative type of glomerulonephritis is characterized by the number of cell in the glomeruli (lack of hypercellularity), divided into three sub-division: Minimal change GN, Focal Segmental Glomeru-
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