Schmiedl A, Lührmann A, Pabst R, Koslowski R
Institute of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany.
Int Arch Allergy Immunol. 2009;148(2):118-26. doi: 10.1159/000155742. Epub 2008 Sep 19.
The surfactant proteins SP-A and SP-D, components of the innate immune system, are involved in host defence.
We tested the hypothesis that ovalbumin (OVA) challenge leads to an upregulation of both proteins in alveolar epithelial type II cells (AEII) and Clara cells and to an enhanced uptake by macrophages.
After sensitization with OVA and heat-killed Bordetella pertussis challenge followed intratracheally with 0.5% OVA on day 13. One day after challenge lung tissue and bronchoalveolar lavage fluid (BALF) of sensitized NaCl- and OVA-challenged Brown Norway rats were compared with home cage controls using qRt-PCR, Western blot and immunohistochemistry.
After OVA challenge (1) eosinophils increased significantly in the BALF, (2) the total amount of SP-A and SP-D was significantly increased in lung tissue, (3) the amount of SP-A was significantly and the amount of SP-D was remarkably elevated in BALF, and (4) the levels of SP-A and SP-D mRNA in lung tissue were significantly elevated. Using quantitative immunohistochemistry, we found (5) significantly higher surface fractions of SP-A- and SP-D-labelled AEII, (6) no differences in the surface fractions of SP-A- and SP-D-labelled bronchial Clara cells, and (7) a significantly increased cell density of unlabelled and SP-A-labelled macrophages.
Thus, combining molecular biological and histological methods we suggest that after OVA challenge (1) AEII but not Clara cells show a significantly higher expression of SP-A and SP-D leading also to higher amounts of both SPs in BALF and (2) macrophages gather predominantly SP-A.
表面活性蛋白A(SP-A)和表面活性蛋白D(SP-D)是固有免疫系统的组成部分,参与宿主防御。
我们检验了以下假设,即卵清蛋白(OVA)激发会导致肺泡II型上皮细胞(AEII)和克拉拉细胞中这两种蛋白的上调,并增强巨噬细胞的摄取。
用OVA致敏并经热灭活百日咳博德特氏菌激发后,于第13天经气管内给予0.5% OVA。激发后一天,使用qRt-PCR、蛋白质免疫印迹和免疫组织化学方法,将致敏的NaCl和OVA激发的棕色挪威大鼠的肺组织和支气管肺泡灌洗液(BALF)与饲养笼对照进行比较。
OVA激发后,(1)BALF中的嗜酸性粒细胞显著增加,(2)肺组织中SP-A和SP-D的总量显著增加,(3)BALF中SP-A的量显著增加,SP-D的量显著升高,(4)肺组织中SP-A和SP-D mRNA的水平显著升高。使用定量免疫组织化学,我们发现(5)SP-A和SP-D标记的AEII的表面分数显著更高,(6)SP-A和SP-D标记的支气管克拉拉细胞的表面分数没有差异,(7)未标记和SP-A标记的巨噬细胞的细胞密度显著增加。
因此,结合分子生物学和组织学方法,我们认为OVA激发后,(1)AEII而非克拉拉细胞显示出SP-A和SP-D的显著更高表达,这也导致BALF中两种表面活性蛋白的量更高,(2)巨噬细胞主要聚集SP-A。