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Cloned endothelium derived from autoimmune vascular disease retain structural and functional characteristics of
C F Moyer1, E Huggins, S Sarantopoulos
1Department of Comparative Medicine, Bowman Gray Medical School, Winston-Salem, North Carolina 27103.
Abstract:
MRL/1pr mice demonstrate anatomic specificity in their development of vasculitis including the small- and medium-sized muscular arteries of the mesentery. To define the functional role of endothelium in vasculitis, we have cloned endothelial cells derived from inflamed small- and medium-sized arteries. Primary cells were derived by enzymatic dispersement and endothelial cells were selected by utilizing a combination of specific culture conditions. Cloned endothelium were developed utilizing limiting dilution cultures supplemented by endothelial cell growth factor. The cloned endothelial cells express many structural features of mature endothelial cells including Factor VIII-RA, non-muscle-specific actin, and Weibel-Palade bodies. Functionally, the clones express functional receptors for the scavenger pathway for LDL metabolism. The cells do not express Class I MHC antigens; however, IFN-beta and IFN-gamma stimulate Class I MHC expression after 24 h, which induces lysis of virus-infected cloned endothelium by Class I-restricted virus-primed T cells. In direct contrast to site-identical vascular smooth muscle cells (VSMCs), endothelial cells do not spontaneously express Class II MHC antigens, nor do they secrete biologically relevant levels of IL-1 unless triggered by lipopolysaccharide. The availability of site-specific cloned endothelium along with cloned VSMCs from autoimmune mice should resolve major experimental controversies involving the pathophysiology of inflammatory vascular disease.
Insights
Researchers cloned endothelial cells from inflamed mouse arteries to study vasculitis. These cells exhibit mature features and specific immune responses, aiding research into inflammatory vascular diseases.
Area of Science:
- Immunology
- Cell Biology
- Pathology
Background:
- MRL/1pr mice develop specific vasculitis in mesenteric arteries.
- The role of endothelium in vasculitis requires further definition.
Purpose of the Study:
- To clone and characterize endothelial cells from inflamed mouse arteries.
- To investigate the functional and structural properties of these cloned endothelial cells.
Main Methods:
- Enzymatic dispersement and selective culture conditions were used to derive primary cells.
- Cloned endothelium was developed using limiting dilution cultures with endothelial cell growth factor.
- Characterization included structural features (Factor VIII-RA, Weibel-Palade bodies) and functional assays (MHC antigen expression, IL-1 secretion).
Main Results:
- Cloned endothelial cells displayed mature endothelial features and functional scavenger receptors for LDL.
- IFN-beta and IFN-gamma induced Class I MHC expression, enabling lysis by T cells.
- Endothelial cells did not spontaneously express Class II MHC or secrete IL-1 without LPS stimulation, unlike VSMCs.
Conclusions:
- Site-specific cloned endothelium and VSMCs from autoimmune mice provide a model to study inflammatory vascular disease.
- This model can help resolve controversies in the pathophysiology of vasculitis.