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Different mechanisms by which anti-DNA MoAbs bind to human endothelial cells and glomerular mesangial cells
T M Chan1, G Frampton, N A Staines
1Renal Unit, UMDS, University of London, UK.
Abstract:
The mechanisms by which anti-DNA MoAbs derived from MRL-lpr/lpr mice, bind to human umbilical vein endothelial cells (HUVEC) and glomerular mesangial cells were studied using a cellular ELISA. DNAse-treatment of either the MoAb or HUVEC followed by reconstitution with DNA and/or histones was performed to determine whether DNA and histones mediated such binding. It was found that MoAb410 bound to HUVEC and mesangial cells in the form of preformed DNA/anti-DNA immune complex, and such binding was facilitated by histones. In contrast, MoAb 152 bound directly to cell membrane-associated DNA, and adding DNA to MoAb 152 reduced its cellular binding. DNA binds endothelial cell surface and histones enhance the binding of both MoAb 410 and MoAb 152 to HUVEC by increasing cell membrane-associated DNA. Finally, the degree of MoAb binding to HUVEC is critically influenced by the relative concentrations of antibody, DNA, and histones.
Insights
Two types of anti-DNA monoclonal antibodies (MoAbs) bind differently to endothelial and mesangial cells. Histones enhance binding by increasing cell-associated DNA, influencing antibody-DNA-histone interactions.
Area of Science:
- Immunology
- Cell Biology
- Nephrology
Background:
- Anti-DNA monoclonal antibodies (MoAbs) are crucial in autoimmune diseases.
- Understanding their binding mechanisms to endothelial and mesangial cells is vital for disease research.
Purpose of the Study:
- To elucidate the distinct binding mechanisms of anti-DNA MoAbs to human umbilical vein endothelial cells (HUVEC) and glomerular mesangial cells.
- To investigate the roles of DNA and histones in mediating this cellular binding.
Main Methods:
- Cellular ELISA was employed to study MoAb binding.
- DNAse treatment and reconstitution with DNA/histones were used to dissect binding components.
Main Results:
- MoAb410 binds HUVEC and mesangial cells via preformed DNA/anti-DNA immune complexes, with histone facilitation.
- MoAb 152 directly binds to cell membrane-associated DNA; excess DNA inhibits this binding.
- Histones enhance binding of both MoAbs to HUVEC by increasing cell membrane-associated DNA.
Conclusions:
- Distinct binding pathways exist for different anti-DNA MoAbs.
- Histones play a significant role in enhancing MoAb binding to endothelial cells.
- Antibody, DNA, and histone concentrations critically influence MoAb binding to HUVEC.