Reactivity of anti-neutrophil cytoplasmic autoantibodies with HL-60 cells.
Anti-neutrophil cytoplasmic autoantibodies (ANCA) react with antigens in the cytoplasm of neutrophils and monocytes, and are found in the blood of patients with necrotizing vasculitis and pauci-immune necrotizing and crescentic glomerulonephritis. The standard techniques for ANCA analysis use human polymorphonuclear leukocytes (PMN) as a source for antigen. A comparison was made between ANCA reactivity with human PMN and HL-60 cells. Fifty-five ANCA-positive and 17 ANCA-negative sera were reacted with HL-60 cells in an indirect immunofluorescence microscopy assay to assess the ability of HL-60 cells to distinguish between positive and negative reactivity and between the two major types of ANCA. The HL-60 cell method agreed with a PMN method in all but one instance. In an enzyme immunoassay, HL-60 primary granules could be used to detect ANCA. Evidence is also presented that the perinuclear staining of HL-60 cells and neutrophils by ANCA with anti-myeloperoxidase specificity is caused by the nucleophilic properties of myeloperoxidase. Reactivity with HL-60 cells further elucidates the antigen specificity of both types of ANCA and indicates that HL-60 cells are a suitable tissue culture cell line for investigating the pathobiology of ANCA.
Charles
L A
LA
Department of Pathology, University of North Carolina School of Medicine, Chapel Hill 27599-7525.
Falk
R J
RJ
Jennette
J C
JC
D001323:Autoantibodies / Q000276:immunology*; D002467:Cell Nucleus / Q000276:immunology; D003593:Cytoplasm / Q000276:immunology; D003594:Cytoplasmic Granules / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D007951:Leukemia, Myeloid / Q000276:immunology*; D009504:Neutrophils / Q000276:immunology*; D009195:Peroxidase / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured
D001323:Autoantibodies; D009195:Peroxidase