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Deposition of kappa and lambda light chains in amyloid filaments of dialysis-related amyloidosis
D Brancaccio1, G M Ghiggeri, P Braidotti
1Renal Unit, Ospedale San Paolo, Milano, Italy.
Abstract:
beta 2-Microglobulin (beta 2m) is considered to be the amyloidogenic precursor in dialysis-related amyloidosis, although the implication of other relevant cofactors in the pathogenesis of this disease has also been hypothesized. It is conceivable that substances found in amyloid deposits might represent something more than simple codeposition, possibly playing a pathogenic role in amyloidogenesis. Along these lines, a detailed analysis of the protein composition of amyloid fibrils purified from synovial material surgically obtained from nine patients on long-term dialysis was carried out. By the use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, several other protein components, in addition to beta 2m, were found. These were characterized by NH2 amino-terminal sequencing and immunoblotting. In fibrils obtained by water extraction, which fulfill the electron microscopy criteria of highly pure amyloid material, polyclonal kappa and lambda light chains were detected with a concentration of 15 micrograms/mL in the water extraction material; the beta 2m concentration was 200 micrograms/mL. Light microscopy immunohistochemistry was performed on samples from five patients. Amyloid deposits reacted with anti-beta 2m, and anti-light (kappa, lambda), chain antibodies. The immunoreaction of amyloid filaments to anti-beta 2m, anti-lambda, and anti-kappa light chain antibodies was also tested by electron microscopy by use of the immunogold staining procedure. Amyloid filaments were labeled by the three antibodies and showed a different intensity of immunostaining apparently related to their different aggregation pattern. These observations demonstrate that polyclonal immunoglobulin light chains (kappa and lambda) are not contaminants but, together with beta 2m, represent a major constituent of amyloid deposits in dialysis-related osteoarticular amyloidosis, thus indicating their possible role in amyloidogenesis.
Insights
Dialysis-related amyloidosis involves beta 2-microglobulin and immunoglobulin light chains. These proteins, not just beta 2-microglobulin, are major components of amyloid deposits, suggesting a role in disease development.
Area of Science:
- Biochemistry
- Immunology
- Pathology
Background:
- Dialysis-related amyloidosis is linked to beta 2-microglobulin (beta 2m).
- The role of other cofactors in the pathogenesis of this condition is hypothesized.
- Amyloid deposits may contain substances beyond simple codeposition that contribute to pathogenesis.
Purpose of the Study:
- To analyze the protein composition of amyloid fibrils in dialysis-related osteoarticular amyloidosis.
- To determine if other proteins besides beta 2m are significant components of these amyloid deposits.
- To investigate the potential pathogenic role of these co-constituents in amyloidogenesis.
Main Methods:
- Purification of amyloid fibrils from synovial material of patients undergoing long-term dialysis.
- Protein analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
- Identification and characterization of proteins via NH2 amino-terminal sequencing, immunoblotting, light microscopy immunohistochemistry, and immunogold electron microscopy staining.
Main Results:
- SDS-PAGE revealed protein components in addition to beta 2m.
- Polyclonal kappa and lambda immunoglobulin light chains were detected in purified amyloid fibrils.
- Immunohistochemistry and immunogold staining confirmed the presence of beta 2m and both kappa and lambda light chains within amyloid deposits and filaments.
Conclusions:
- Polyclonal immunoglobulin light chains (kappa and lambda) are major constituents of amyloid deposits in dialysis-related osteoarticular amyloidosis.
- These light chains, along with beta 2m, are not mere contaminants but play a significant role.
- The findings suggest a potential pathogenic role for immunoglobulin light chains in the process of amyloidogenesis in this condition.