| Anti-neutrophil cytoplasmic antibodies (ANCA) are one kind of antibodies aimed at the main granule components in neutrophils and lysosomes in monocytes.It is divided into cytoplasmic -ANCA (C-ANCA) and perinuclear-ANCA (P-ANCA) according to the different staining region by indirect irnmunofluorescence detection. The main target antigen of C-ANCA is proteinase 3 (PR-3), and P-ANCA is myeloperoxidase (MPO). As one kind of specific serum antibodies, ANCA was regarded as of great importance in the predicting of remission and relapse for systemic necrotic vasculitis, of which Wegener's Granulomatosis (WG) is thought to be the representative disease. At the same time, ANCA was found in many other autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, dermatomyositis, etc, so that it is by now considered to have great correlation with not only systemic vasculitis but also autoimmune diseases.Patients and methodsWe investigated 122 patients including 84cases with connective tissue diseases and 38 cases with vascular dermatoses from January 2003 to January 2004. 36 cases in connective tissue disease group were with SLE (19 of which are renal involvement), 16 cases with rheumatoid arthritis, 12 cases withdermatomyositis, 8 cases with scleroderma, 8 cases with Sjogren syndrome, 3 cases with overlap syndrome, and 1 case with Mixed Connective Tissue Disease. 14 cases in vascular dermatoses group were with anaphylactoid purpura, 12 cases with erythema nodosum, 7 cases with allergic cutaneous vasculitis, and 5 cases with Bechet's disease. All patients met the diagnostic criteria of Clinical Dermatology edited by ZHAO Bian. 30 cases volunteers were recruited as healthy control.ANCA was detected by using indirect immunofluorescence (IIF) test. The antibodies kit was purchased from Euroimmun Company (Germany). Diluted serum samples with phosphate-buffered saline (PBS-Tween), applied 25ul of diluted serum to each reaction field of the reagents' support. Incubated for 30 min at room temperature. Rinsed the slides with a flush of PBS-Tween to get rid of unconjugated samples and then immersed them immediately afterwards into a cuvette which contains PBS-Tween for at least 5 min. Dropped 20 ul of fluorescein-labelled anti-human globulin onto each reaction field of a clean reagents' support and incubated for 30 min at room temperature were protected. The slides from direct sunlight. Rinsed the slides with a flush of PBS-Tween to get rid of unconjugated samples and then immersed them immediately afterwards into a cuvette which contains PBS-Tween for at least 5 min. Rinsed again, then placed glycerol drop of 10 ul onto a cover glass per reaction field and read the fluorescence with the microscope. Specific green fluorescence in granulocytes was considered as positive reaction. The positive result was divided into +~+++ grades according to the strength of fluorescence. Two relevant fluorescence patterns can be differentiated: C-ANCA showed a granular fluorescence which is distributed regularly over the entire cytoplasm of the granulocytes, and P-ANCA showed a smooth fluorescence wrapped ribbon-like around the cell nuclei of the granulocytes.Results and discussionsAll cases with positive ANCA demonstrated P-ANCA pattern. 46.4% ofconnective tissue disease cases were ANCA positive and vascular dermatose cases 13.2%. All cases in healthy control group were found ANCA negative, showing a significant difference comparing with the two formers groups(P<0.001 and P<0.05).In the connective tissue diseases group (84 cases in all), 20 out of 36 SLE patients were ANCA positive (positive rate 55.6%); 8 out of 16 patients with rheumatoid arthritis were ANCA positive (positive rate 50%), and the patients with positive reaction remarkably had much more serious and active disease condition as well as longer disease duration ; 2 out of 12 patients with dermatomyositis were ANCA positive (positive rate 16.7%); 2 out of 8 patients with progressive systemic scleroderma were ANCA positive (p...
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