The assessment of anti-endothelial cell antibodies in scleroderma-associated pulmonary fibrosis: A study of indirect immunofluorescent and Western blot analysis in 49 patients with scleroderma
Fluorescent Antibody Technique, Indirect
We recently reported on the use of an indirect immunofluorescent method designated the rodent lung assay; this test assesses for the presence of circulating antibodies directed at components of the microvasculature. Serum samples from 49 patients with scleroderma were incubated with rodent lung tissue sections and visualized with fluoresceinated human anti-IgG. The assay also was performed on samples from a control group. Western blot analysis was performed with endothelial cell protein extracts using serum samples from patients with scleroderma and from healthy control subjects. The control subjects had a negative indirect immunofluorescent assay result. In the patients with scleroderma, there was a significant positive correlation between intensity of indirect immunofluorescent staining and pulmonary fibrosis (r = 0.316; P = .0347) and hypertension (r = 0.310; P = .0408). Western blot analysis revealed antibody binding to proteins in extracts of human endothelial cells in all patients in whom there was evidence of pulmonary disease. The indirect immunofluorescent rodent lung assay and Western blot data support a potential role of anti-endothelial cell antibodies in the propagation of scleroderma-associated pulmonary disease.