Lab of Plant Development, Shanghai Key Laboratory of Bio-Energy Crops, Plant Science Center, School of Life Sciences, Shanghai University, Shanghai, 200444, China.
School of Environmental and Chemical Engineering, Shanghai University, Shanghai, 200444, China.
Biochem Biophys Res Commun. 2019 Jun 11;513(4):767-774. doi: 10.1016/j.bbrc.2019.03.210. Epub 2019 Apr 13.
Pollen allergy is a very serious seasonal respiratory disease. However, there has been a lack of understanding how pollen allergens enter the body and act on cells. This study focused on the release, transport and characteristic of Pla a3 allergen of the Platanus acerifolia pollen. Pla a3 protein was purified by prokaryotic expression system for preparation of polyclonal antibody. The distribution and release of Pla a3 protein in pollen were observed by immunohistochemistry. Mice were immunized with purified Pla a3 protein and SPPs, respectively. The pathological examination of mouse lung tissue proved that SPPs, as a fine particle in the range of 0.1-1μm, can enter the deep part of the lung directly through the respiratory tract and led to inflammation. Furthermore, DAPI staining confirmed a certain amount of nucleic acids in SPPs. After incubation with SPPs for 6 h, the Pla a3 mRNA could be detected in A549 cells by PCR. This suggests that nucleic acid wrapped in SPPs could be delivered into A549 cells. These results could provide a new clue and experimental data accumulation for further study on the mechanism of pollen sensitization.
花粉过敏症是一种非常严重的季节性呼吸道疾病。然而,人们对于花粉过敏原如何进入人体并作用于细胞还缺乏了解。本研究专注于悬铃木花粉 Pla a3 过敏原的释放、运输和特性。通过原核表达系统对 Pla a3 蛋白进行了纯化,以制备多克隆抗体。通过免疫组织化学观察 Pla a3 蛋白在花粉中的分布和释放。分别用纯化的 Pla a3 蛋白和 SPPs 对小鼠进行免疫。对小鼠肺组织的病理检查证明,SPPs 作为粒径在 0.1-1μm 范围内的细小颗粒,可以通过呼吸道直接进入肺部深处,导致炎症。此外,DAPI 染色证实 SPPs 中含有一定量的核酸。用 SPPs 孵育 6 小时后,PCR 可检测到 A549 细胞中的 Pla a3 mRNA。这表明包裹在 SPPs 中的核酸可以递送到 A549 细胞中。这些结果为进一步研究花粉致敏的机制提供了新的线索和实验数据积累。