Urine levels of CD46 (membrane cofactor protein) are increased in patients with glomerular diseases.
Soluble membrane cofactor protein (MCP, CD46) has not been detected by conventional ELISA in human urine. Here, we established a highly sensitive assay method for determination of urinary MCP (uMCP) using monoclonal antibody-coated paramagnetic beads. This method enabled us to detect less than 0.05 ng/ml of purified membrane and recombinant soluble MCP, a sensitivity 10-fold higher than that of conventional ELISA. In normal subjects, the levels of uMCP were <0. 05 ng/ml. The levels of uMCP were elevated in patients with IgA nephropathy and more prominently in patients with rapidly progressive glomerulonephritis. The levels of uMCP were correlated significantly with those of serum MCP (sMCP) and N-acetyl-beta-glucosaminidase and nonsignificantly with those of beta(2)-microglobulin, total urine protein, or serum creatinine. The properties of uMCP were inconsistent with those of the reported sMCP, since uMCP showed three bands on SDS-PAGE/immunoblotting with molecular mass profiles different from those of sMCP. uMCP exhibited factor I cofactor activity for cleavage of C3b comparable to that of sMCP. The origin of uMCP, however, remains to be determined. These results, taken together with the parameter correlation profiles, suggested that uMCP is secreted or produced secondary to tubular or glomerular damage. The physiological role and clinical significance of uMCP are now within the scope of our investigation by establishment of this assay.
Shoji
T
T
Department of Nephrology, Osaka Prefectural Hospital, Sumiyoshi-ku, Osaka, Japan.
Nakanishi
I
I
Kunitou
K
K
Tsubakihara
Y
Y
Hirooka
Y
Y
Kishi
Y
Y
Hatanaka
M
M
Matsumoto
M
M
Toyoshima
K
K
Seya
T
T
D000118:Acetylglucosaminidase / Q000097:blood / Q000652:urine; D015703:Antigens, CD / Q000097:blood / Q000737:chemistry / Q000652:urine*; D000990:Antithrombin III / Q000378:metabolism; D003179:Complement C3b / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005921:Glomerulonephritis / Q000097:blood / Q000652:urine*; D005922:Glomerulonephritis, IGA / Q000097:blood / Q000652:urine*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000097:blood / Q000737:chemistry / Q000652:urine*
D015703:Antigens, CD; C497857:CD46 protein, human; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D003179:Complement C3b; D000990:Antithrombin III; D000118:Acetylglucosaminidase