Related Experiment Videos
Peritoneal mesothelial cells produce complement factors and express CD59 that inhibits C5b-9-mediated cell lysis
G Barbano1, F Cappa, I Prigione
1Nephrology Department, G. Gaslini Children's Hospital, Genoa, Italy.
Insights
Mesothelial cells (MCs) express CD59, protecting them from complement-mediated lysis during chronic peritoneal dialysis (CPD). MCs also produce complement components, influencing peritoneal cavity health.
Area of Science:
- Immunology
- Cell Biology
- Nephrology
Background:
- Complement (C) activation and C5b-9 generation occur in the peritoneal cavity during chronic peritoneal dialysis (CPD).
- Mesothelial cells (MCs) are crucial for peritoneal membrane integrity.
- The role of CD59 and complement production by MCs in CPD is not fully understood.
Purpose of the Study:
- To investigate CD59 expression on MCs.
- To examine complement component production by MCs.
- To determine the functional significance of CD59 on MCs in the context of complement-mediated lysis.
Main Methods:
- Cultured MC lines from children on CPD and non-uremic children.
- Immunohistochemical staining using anti-CD59 monoclonal antibody (mAb).
- Western blotting to confirm CD59 presence.
- Assessment of MC cytotoxicity after incubation with anti-CD59 mAb.
- Detection of complement components in MC supernatants using immunoassays.
Main Results:
- All MC lines expressed CD59 on their surface.
- Western blotting confirmed the presence of CD59 in MC membranes.
- Blocking CD59 with anti-CD59 mAb resulted in 100% MC lysis.
- MCs produced C3, C4, and C6; non-uremic MCs also produced C5, C7, C8, and C9, with higher C4 concentrations.
Conclusions:
- CD59 expression on MCs provides protection against C5b-9-mediated lysis.
- MCs are capable of producing various complement components.
- These findings suggest a role for MC CD59 and complement production in protecting the peritoneal membrane from infection and damage during CPD.
Abstract:
The CD59 membrane protein confers protection from C5b-9-mediated cell lysis. Because evidence exists for complement (C) activation and generation of C5b-9 in the peritoneal cavity during chronic peritoneal dialysis (CPD), we investigated, on mesothelial cell (MC) lines, the expression of CD59 and the production of C components. Four MC lines were obtained from children on CPD, and two from non uremic children. CD59 expression on MCs was investigated with anti-CD59 monoclonal antibody (mAb) and polyclonal goat immunoglobulin G (IgG). MC lines were positive for staining with anti-CD59 mAb. Western blotting analysis of MC membrane demonstrated a band with the same molecular weight as CD59. Incubation of MC with anti-CD59 mAb abrogated the protective effect of CD59 (100% cytotoxicity). C3, C4, and C6 were detected in the supernatants of MC; in non uremic MC supernatants, C5, C7, C8, and C9 were also detectable, and C4 concentration was tenfold higher. CD59 expression confers to MCs protection from C5b-9-mediated lysis. MCs produce C factors. These findings suggest that production of complement components and expression of CD59 on MCs could play a role both in peritoneal cavity infection (decreased complement production) and in peritoneal membrane damage (decreased CD59 expression and reduced remesothelialization owing to MC lysis).