Major histocompatibility complex class II (DR) antigen and costimulatory molecules on in vitro and in vivo activated human polymorphonuclear neutrophils
Major histocompatibility complex class II (DR) antigen and costimulatory molecules on in vitro and in vivo activated human polymorphonuclear neutrophils
复制标题
DOI：
10.1111/j.1365-2567.2006.02471.x
复制
发表时间：
2006-12-01
期刊：
影响因子：
6.4
通讯作者：
Baxter, Derek
中科院分区：
文献类型：
--
作者：
Sandilands, Gavin P.;McCrae, Jame;Baxter, Derek
我们之前的研究表明，正常的人外周血多形核中性粒细胞（pmn）含有细胞质中通常与抗原呈递和t细胞共刺激相关的三种关键分子，即主要组织相容性复合体II类（DR）抗原，CD80 （B7-1）和CD86 （B7-2）。发现这些细胞质分子在Mac-1交联（X-L）后几分钟内转移到细胞表面，Mac-1是一种早期中性粒细胞激活信号。在这项研究中，我们比较了Mac -1的X-L与其他四种有充分记录的体外中性粒细胞激活剂：肉豆肉酸酯、n -甲硫酰基亮基苯丙氨酸、脂多糖和免疫球蛋白G-Latex颗粒的吞噬作用。此外，我们使用从外周血（作为对照）和类风湿关节炎患者的滑液中获得的中性粒细胞配对样本作为体内活化细胞的来源。除吞噬作用外，根据流式细胞术测量的大小、粒度和表型，所有激活因子都能快速（在30分钟内）产生两群活化的中性粒细胞（指定为P1和P2）。DR和共刺激分子显著上调，主要在P2细胞上，除吞噬作用外，所有激活因子均显著上调。我们注意到CD80和CD86以不同的模式响应各种激活信号，这表明它们的细胞内颗粒位置可能不同。双染色共聚焦激光显微镜研究表明，CD80主要局限于分泌囊泡（SVs），而CD86似乎在SVs和继发性（特异性）和原发性（亲氮性）颗粒中有更广泛的分布。在共聚焦激光显微镜下观察到，这些抗原的表面表达也在P2滑液中性粒细胞上增加，在细胞表面呈大的异质团簇。
We have previously shown that normal human peripheral blood polymorphonuclear neutrophils (PMNs) contain cytoplasmic 'stores' of three key molecules normally associated with antigen presentation and T-cell costimulation, i.e. major histocompatibility complex class II (DR) antigen, CD80 (B7-1) and CD86 (B7-2). These cytoplasmic molecules were found to translocate to the cell surface within a few minutes following cross-linking (X-L) of Mac-1: an early neutrophil activation signal. In this study we have compared X-L of Mac -1 in parallel with four other well documented in vitro neutrophil activators: phorbol myristate acetate, N-formyl methionyl leucyl phenylalanine, lipopolysaccharide, and phagocytosis of immunoglobulin G-Latex particles. In addition, we have used paired samples of neutrophils obtained from peripheral blood (as a control) and synovial fluid from patients with rheumatoid arthritis as a source of in vivo activated cells. With the exception of phagocytosis, all activators resulted in the rapid (within 30 min) generation of two populations of activated neutrophils (designated P1 and P2) based on flow-cytometry measurements of size, granularity and phenotype. Significant up-regulation of DR and costimulatory molecules was observed, predominantly on P2 cells, with all activators except phagocytosis. CD80 and CD86 were noted to respond to the various activation signals in a different pattern suggesting that their intracellular granule location may be different. Dual-staining confocal laser microscopy studies showed that CD80 is largely confined to secretory vesicles (SVs) while CD86 appears to have a much wider distribution being found in SVs and within secondary (specific) and primary (azurophilic) granules. Increased surface expression of these antigens was also observed on P2 synovial fluid neutrophils appearing as large heterogeneous clusters on the cell surface when visualized by confocal laser microscopy.