Department of Immunology-Allergology-Rheumatology, Faculty of Medicine, University of Antwerp, Antwerp, Belgium.
Cytometry B Clin Cytom. 2010 Sep;78(5):302-7. doi: 10.1002/cyto.b.20531. Epub 2010 May 7.
P38 mitogen-activated protein kinase (MAPK) is known to govern IgE-mediated basophil activation. Intracellular phosphorylated p38 MAPK (Pp38 MAPK) in IgE-activated basophils can be quantified flow cytometrically.
To study whether Pp38 MAPK constitutes a potential novel read-out for flow-assisted diagnosis of hymenoptera venom allergy and to investigate whether this marker allows follow-up of successful venom immunotherapy (VIT).
Fifty-two patients with documented wasp venom allergy and seven wasp-stung asymptomatic control individuals were enrolled. Wasp venom-induced basophil activation was analyzed flow cytometrically with anti-IgE, anti-CD63, and anti-Pp38 MAPK to assess their activation status before starting immunotherapy. To assess whether p38 MAPK constitutes a candidate marker for monitoring VIT, we repeated the basophil activation test (BAT) in 25 patients on the fifth day of a build-up immunotherapy. In addition, we investigated whether the Pp38 MAPK-based BAT could contribute in the decision of discontinuing VIT in a cross-sectional analysis in 13 patients receiving treatment for 3 years and 14 patients receiving treatment for 5 years.
Patients exhibited a dose-dependent basophil activation with phosphorylation of p38 MAPK and upregulation of downstream CD63. In contrast, stung controls demonstrated a dose-dependent but "abrogated" signal transduction in basophils with less and shorter duration of the phosphorylation of p38 MAPK and without subsequent upregulation of CD63. When repeated after 5 days of VIT and when investigated cross-sectionally after 3 years or 5 years of maintenance therapy, no effect of VIT on the phosphorylation of p38 MAPK was demonstrable.
This study discloses that not only basophils from patients, but also from the stung control individuals, respond to wasp venom stimulation with phosphorylation of p38 MAPK, although to a lesser extend. No clear effect of VIT on the phosphorylation of p38 MAPK was shown. Thus, although p38 MAPK provides an additional tool in the diagnosis of wasp venom allergy, it does not contribute to the decision whether to stop successful VIT. © 2010 International Clinical Cytometry Society.
已知 P38 丝裂原活化蛋白激酶（MAPK）可调控 IgE 介导的嗜碱性粒细胞激活。通过流式细胞术可定量检测 IgE 激活的嗜碱性粒细胞中的细胞内磷酸化 p38 MAPK（Pp38 MAPK）。
研究 Pp38 MAPK 是否可作为一种潜在的新型检测指标，用于辅助诊断蜂类毒液过敏，并探讨该标志物是否可用于随访成功的蜂毒免疫治疗（VIT）。
纳入 52 例有蜂类毒液过敏史的患者和 7 例被蜂蜇无症状的对照个体。采用抗 IgE、抗 CD63 和抗 Pp38 MAPK 进行流式细胞术分析，以评估开始免疫治疗前的嗜碱性粒细胞激活状态。为评估 p38 MAPK 是否可作为监测 VIT 的候选标志物，我们在 25 例患者接受递增免疫治疗的第 5 天重复了嗜碱性粒细胞激活试验（BAT）。此外，我们还通过对接受治疗 3 年和 5 年的 13 例和 14 例患者进行横断面分析，研究了基于 Pp38 MAPK 的 BAT 是否有助于决定是否停止 VIT。
患者表现出剂量依赖性的嗜碱性粒细胞激活，伴有 p38 MAPK 的磷酸化和下游 CD63 的上调。相比之下，被蜂蜇的对照个体表现出剂量依赖性但“被阻断”的信号转导，p38 MAPK 的磷酸化程度较低且持续时间较短，随后 CD63 也没有上调。在 VIT 治疗 5 天后重复该试验，以及在维持治疗 3 年或 5 年后进行横断面研究时，均未显示 VIT 对 p38 MAPK 磷酸化有影响。
本研究揭示，不仅是患者的嗜碱性粒细胞，而且是被蜂蜇的对照个体的嗜碱性粒细胞，对蜂类毒液刺激均会发生 p38 MAPK 的磷酸化反应，尽管程度较轻。VIT 对 p38 MAPK 的磷酸化无明显影响。因此，尽管 p38 MAPK 为蜂类毒液过敏的诊断提供了另一种工具，但它不能用于决定是否停止成功的 VIT。
© 2010 国际临床细胞化学学会。
