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Urine levels of CD46 (membrane cofactor protein) are increased in patients with glomerular diseases
T Shoji1, I Nakanishi, K Kunitou
1Department of Nephrology, Osaka Prefectural Hospital, Sumiyoshi-ku, Osaka, Japan.
Abstract:
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.
Insights
A new, highly sensitive assay detects urinary soluble membrane cofactor protein (MCP, CD46) in human urine. Elevated levels correlate with kidney disease, suggesting MCP
Area of Science:
- Nephrology
- Immunology
- Biochemistry
Background:
- Soluble membrane cofactor protein (MCP, CD46) is crucial for immune regulation.
- Conventional ELISA methods fail to detect MCP in human urine.
- The presence and significance of urinary MCP (uMCP) remain unclear.
Purpose of the Study:
- To develop a sensitive assay for quantifying urinary MCP (uMCP).
- To investigate the levels and clinical relevance of uMCP in kidney diseases.
Main Methods:
- Development of a highly sensitive assay using monoclonal antibody-coated paramagnetic beads.
- Detection of MCP with a sensitivity of <0.05 ng/ml, 10-fold higher than conventional ELISA.
- Analysis of uMCP levels in healthy subjects and patients with IgA nephropathy and rapidly progressive glomerulonephritis.
Main Results:
- Established a sensitive assay for uMCP detection (<0.05 ng/ml).
- Observed elevated uMCP levels in IgA nephropathy and rapidly progressive glomerulonephritis patients.
- Found significant correlations between uMCP, serum MCP, and N-acetyl-beta-glucosaminidase.
Conclusions:
- The novel assay enables sensitive detection of uMCP.
- Elevated uMCP levels are associated with specific kidney diseases.
- uMCP may be a biomarker for tubular or glomerular damage, warranting further investigation into its origin and clinical significance.