Johns Hopkins Asthma and Allergy Center, 5501 Hopkins Bayview Circle, Baltimore, MD 21224, USA.
Clin Exp Allergy. 2010 Sep;40(9):1365-77. doi: 10.1111/j.1365-2222.2010.03572.x. Epub 2010 Jul 13.
Activation of human basophils results in the release of many different mediators and the expression of new cell surface proteins. The markers CD63 and CD203c have been used in recent years to assess basophil activation but there have been many studies that demonstrate that expression of these markers can be dissociated from histamine release.
To determine the signal transduction requirements for CD203c and CD63 expression.
The current study began by exploring the dependency of CD203c and CD63 expression on protein kinase C (PKC) using known selective inhibitors of PKC.
Between 30 and 300 nm, Ro-31-8220 and bisindoylmaleimide II (Bis II) had no effect on formyl-met-leu-phe- or anti-IgE-induced CD63 or CD203c but enhanced IgE-mediated expression of CD63 by an average of 15-fold at concentrations >1 microm. These results led to the suggestion that these inhibitors altered the normal pathways of degranulation (by a non-PKC dependent mechanism), shifting the normal presence of piecemeal degranulation to the process termed anaphylactic degranulation (AND). Morphological studies demonstrated that concentrations of Ro-31-8220 and Bis II>1 mum dramatically increased the presence of degranulation sacs, a morphological feature of AND.
It is proposed that CD63 expression results from only the AND form of histamine release.
人类嗜碱性粒细胞的激活会导致许多不同介质的释放和新的细胞表面蛋白的表达。近年来,CD63 和 CD203c 标志物已被用于评估嗜碱性粒细胞的激活,但有许多研究表明,这些标志物的表达可以与组胺释放分离。
确定 CD203c 和 CD63 表达的信号转导要求。
本研究首先通过使用已知的 PKC 选择性抑制剂探索 CD203c 和 CD63 表达对蛋白激酶 C(PKC)的依赖性。
在 30 至 300nm 之间,Ro-31-8220 和双吲哚马来酰亚胺 II(Bis II)对甲酰基-甲硫氨酸-亮氨酸-苯丙氨酸或抗 IgE 诱导的 CD63 或 CD203c 没有影响,但在浓度>1μm 时平均增强了 IgE 介导的 CD63 表达 15 倍。这些结果表明,这些抑制剂改变了正常的脱颗粒途径(通过非 PKC 依赖的机制),将正常的碎片脱颗粒转变为称为过敏脱颗粒(AND)的过程。形态学研究表明,浓度为 Ro-31-8220 和 Bis II>1μm 时,大大增加了脱颗粒囊泡的存在,这是 AND 的一种形态特征。
提出 CD63 表达仅来自组胺释放的 AND 形式。