ENDOTHELIAL-CELLS AND RENAL EPITHELIAL-CELLS DO NOT EXPRESS THE WEGENERS AUTOANTIGEN, PROTEINASE-3

Proteinase 3 (PR3) is the major antigen for autoantibodies (C-ANCA) against cytoplasmic components of neutrophils which are strongly associated with Wegener's granulomatosis (WG). Recent data that PR3 may be expressed by renal tubular epithelial cells and endothelial cells suggest potential for a direct pathogenic effect against these cells by C-ANCA or cytoxic T lymphocytes. Using a semi-quantitative polymerase chain reaction.(PCR), ELISA and indirect immunofluorescence staining we studied endothelial and epithelial cell PR3 expression. By PCR, no PR3 expression was found in human umbilical vein endothelial cells (HUVEC) either untreated, or when treated with interferon-gamma (IFN-gamma) (200 U/ml, 6 h, 24 h), IL-I (20 U/ml, 6 h), tumour necrosis factor-alpha, (TNF-alpha) (200 U/ml, 0, 1, 2, 4, 6 h) or IFN-gamma + TNF-alpha (6 h); iliac vein and artery endothelial cells did not express PR3 either. In contrast, PR3 was detected in HL60 cells and neutrophils by PCR, expression being confirmed by sequence analysis. Three PR3 MoAbs showed no binding to unstimulated or TNF-alpha-stimulated HUVEC either by ELISA or by indirect immunofluorescence staining. The epithelial cell line A549 expressed PR3 when assayed by PCR. However, three renal epithelial cell lines (two tubular and one glomerular) showed little or no PR3 expression by PCR or ELISA. These studies fail to demonstrate evidence for PR3 expression by endothelial cells, even when using the highly sensitive PCR assay. Whilst PR3 expression by A549 cells is intriguing, the relevance of this in the pathology of WG is doubtful considering the negligible expression by renal epithelial cell lines.