Japanese Red Cross, Hokkaido Red Cross Blood Center, Sapporo, Japan.
Vox Sang. 2010 May;98(4):560-6. doi: 10.1111/j.1423-0410.2010.01311.x. Epub 2010 Feb 11.
The activation of neutrophils by human leukocyte antigen (HLA) Class I alloantibody is thought to be involved in transfusion-related acute lung injury. Neutrophils contain various biological substances in four groups of granules, including secretory vesicles, azurophilic granules, specific granules and gelatinase granules. To characterize the activation of neutrophils by HLA Class I alloantibody, we investigated whether HLA Class I alloantibody could cause the degranulation of these groups of granules either coordinately or selectively.
Sera containing HLA-A24 alloantibody were incubated with neutrophils in a washed whole blood system. CD11b expression (secretory vesicles) on neutrophils was analysed by flow cytometry, and the secretion of markers of each granule was determined by ELISA.
The treatment of cross-matching-positive neutrophils with sera containing HLA-A24 alloantibody caused the significant expression of CD11b, and the significant secretion of neutrophil elastase and myeloperoxidase, azurophilic granule markers and heparin-binding protein (HBP), which is localized in secretory vesicles and azurophilic granules when compared with cross-matching-negative neutrophils. In contrast, no significant differences were observed in the secretion of lactoferrin, a marker of specific granules, and matrix methalloproteinase-9, a marker of gelatinase granules between cross-matching-positive and cross-matching-negative cells upon stimulation with sera. CD11b expression and secretion of HBP by serum was partially inhibited by p38 mitogen-activated protein (MAP)-kinase inhibitors.
Neutrophils activated with sera containing HLA Class I alloantibody caused the preferential degranulation of azurophilic granules and secretory vesicles. This process was at least in part mediated by p38 MAP kinase-involved signal transduction.
人类白细胞抗原（HLA）I 类同种抗体激活中性粒细胞被认为与输血相关的急性肺损伤有关。中性粒细胞在四种颗粒中包含各种生物物质，包括分泌囊泡、嗜天青颗粒、特异性颗粒和明胶酶颗粒。为了描述 HLA I 类同种抗体对中性粒细胞的激活作用，我们研究了 HLA I 类同种抗体是否可以协同或选择性地引起这些颗粒群的脱颗粒作用。
用含有 HLA-A24 同种抗体的血清孵育洗涤全血系统中的中性粒细胞。通过流式细胞术分析中性粒细胞上 CD11b 的表达（分泌囊泡），并通过 ELISA 测定每个颗粒的分泌标志物。
与含有 HLA-A24 同种抗体的血清孵育交叉匹配阳性的中性粒细胞会导致 CD11b 的显著表达，以及中性粒细胞弹性蛋白酶和髓过氧化物酶、嗜天青颗粒标志物和肝素结合蛋白（HBP）的显著分泌，与交叉匹配阴性的中性粒细胞相比。相比之下，在刺激血清后，特异性颗粒标志物乳铁蛋白和明胶酶颗粒标志物基质金属蛋白酶-9的分泌在交叉匹配阳性和阴性细胞之间没有观察到显著差异。CD11b 表达和 HBP 的分泌被 p38 有丝分裂原激活蛋白（MAP）激酶抑制剂部分抑制。
含有 HLA I 类同种抗体的血清激活的中性粒细胞导致嗜天青颗粒和分泌囊泡的优先脱颗粒。该过程至少部分由 p38 MAP 激酶参与的信号转导介导。
