Onodera Rie, Kurita Emi, Taniguchi Kikuyo, Karakawa Shuhei, Okada Satoshi, Kihara Hirotaka, Fujii Teruhisa, Kobayashi Masao
Department of Pediatrics, Hiroshima University Graduate School of Biomedical & Health Sciences, Hiroshima, Japan.
Department of Medical Technology, Sanyo Women's College, Hatsukaichi, Japan.
Transfusion. 2017 Nov;57(11):2586-2594. doi: 10.1111/trf.14291. Epub 2017 Aug 20.
Anti-human neutrophil antigen (HNA) antibodies have been implicated in the development of neonatal alloimmune neutropenia (NAN) and autoimmune neutropenia (AIN). There are many conventional assay methods that detect anti-HNA antibodies. However, a method to measure multiple samples and detect several anti-HNA antibodies simultaneously is needed.
We developed a new method, the extracted granulocyte antigen immunofluorescence assay (EGIFA), to analyze anti-HNA-1a, -1b, and -2 antibodies in sera. The results obtained by EGIFA were evaluated in comparison with those from several standard assay methods. Anti-HNA antibodies in serum samples from nine familial cases with suspected NAN (n = 19) and children with suspected AIN (n = 88) were also measured by EGIFA.
The evaluation of nine serum samples with anti-HNA antibodies suggested that EGIFA demonstrated equivalent specificity and superior sensitivity to monoclonal antibody-specific immobilization of granulocyte antigens and had comparable sensitivity to the granulocyte indirect immunofluorescence test. EGIFA successfully detected anti-HNA-1a or -1b antibodies in seven of nine familial cases with suspected NAN. EGIFA detected anti-HNA antibodies in 40.9% of children with suspected AIN. Among them, isolated anti-HNA-1a or -1b antibody was detected in 4.5 or 12.5% of children, respectively, and anti-HNA-2 antibody was identified in 3.4% of children. The 30.8% (16 of 52) of children negative for anti-HNA antibody by EGIFA were positive for anti-HLA antibody.
EGIFA facilitated the measurement of anti-HNA-1a, -1b, and/or -2 antibodies in sera. The prompt measurement of anti-HNA antibodies will improve the diagnosis and clinical management of patients with suspected NAN or AIN.
抗人中性粒细胞抗原（HNA）抗体与新生儿同种免疫性中性粒细胞减少症（NAN）和自身免疫性中性粒细胞减少症（AIN）的发生有关。有许多传统检测方法可检测抗HNA抗体。然而，需要一种能同时检测多个样本并检测多种抗HNA抗体的方法。
我们开发了一种新方法，即提取粒细胞抗原免疫荧光测定法（EGIFA），用于分析血清中的抗HNA-1a、-1b和-2抗体。将EGIFA获得的结果与几种标准检测方法的结果进行比较评估。还通过EGIFA检测了9例疑似NAN的家族性病例（n = 19）和疑似AIN的儿童（n = 88）血清样本中的抗HNA抗体。
对9份含抗HNA抗体的血清样本的评估表明，EGIFA显示出与粒细胞抗原单克隆抗体特异性固定法相当的特异性和更高的敏感性，并且与粒细胞间接免疫荧光试验具有相当的敏感性。EGIFA在9例疑似NAN的家族性病例中的7例中成功检测到抗HNA-1a或-1b抗体。EGIFA在40.9%的疑似AIN儿童中检测到抗HNA抗体。其中，分别在4.5%或12.5%的儿童中检测到孤立的抗HNA-1a或-1b抗体，在3.4%的儿童中鉴定出抗HNA-2抗体。EGIFA检测抗HNA抗体阴性的儿童中有30.8%（52例中的16例）抗HLA抗体呈阳性。
EGIFA有助于检测血清中的抗HNA-1a、-1b和/或-2抗体。快速检测抗HNA抗体将改善疑似NAN或AIN患者的诊断和临床管理。
