Wheeler F B, Saluta G, Wise C M, Semble E L, Pisko E J
Department of Medicine, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, NC 27103.
Clin Exp Rheumatol. 1991 Nov-Dec;9(6):569-80.
IgG autoantibodies against antigen in the cytoplasm of cells of the neutrophil-monocyte cell lineage have been found in the sera of patients with Wegener's granulomatosis (WG). The indirect immunofluorescence test (IFT) is proving to be a valuable screening test for these antibodies, but obtaining neutrophils for substrate is time-consuming, and interpretation of the fluorescence patterns in ethanol-fixed cells requires considerable experience. We report an improved IFT using HL-60 cells as substrate. The myeloid reactivity of HL-60 cells was characterized and compared to that of neutrophils, with and without prior ethanol fixation. In contrast to neutrophils, myeloperoxidase (MPO) was more completely extracted from HL-60 cells by prior ethanol fixation, eliminating the confusion inherent in trying to distinguish anti-MPO antibodies from Wegener's granulomatosis associated anti-neutrophil cytoplasmic autoantibodies (WG-ANCA) in the IFT. The WG-ANCA reactivity remained intact with ethanol fixation, producing a distinct crescent and half-moon pattern of specific immunofluorescence. This WG-ANCA positive pattern was found in 25 sera from 11 WG patients and was absent in over 1200 control sera from patients referred for autoantibody testing.
在韦格纳肉芽肿（WG）患者的血清中发现了针对中性粒细胞 - 单核细胞系细胞质中抗原的IgG自身抗体。间接免疫荧光试验（IFT）已被证明是检测这些抗体的一种有价值的筛查试验，但获取用于底物的中性粒细胞很耗时，并且对乙醇固定细胞中荧光模式的解读需要相当多的经验。我们报告了一种以HL - 60细胞为底物的改良IFT。对HL - 60细胞的髓系反应性进行了表征，并与中性粒细胞的髓系反应性进行了比较，包括有无预先乙醇固定的情况。与中性粒细胞不同，预先乙醇固定能更完全地从HL - 60细胞中提取髓过氧化物酶（MPO），消除了在IFT中区分抗MPO抗体与韦格纳肉芽肿相关抗中性粒细胞胞浆自身抗体（WG - ANCA）时固有的混淆。乙醇固定后，WG - ANCA反应性保持完整，产生独特的新月形和半月形特异性免疫荧光模式。在11例WG患者的25份血清中发现了这种WG - ANCA阳性模式，而在转介进行自身抗体检测的患者的1200多份对照血清中未发现。
