Phelps David S, Umstead Todd M, Silveyra Patricia, Hu Sanmei, Wang Guirong, Floros Joanna
C enter for H ost defense, I nflammation, and L ung D isease (CHILD) Research and Departments of Pediatrics, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA.
C enter for H ost defense, I nflammation, and L ung D isease (CHILD) Research and Departments of Pediatrics, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA. ; Obstetrics and Gynecology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA.
J Proteom Genom Res. 2013 Apr 1;1(2):2-26. doi: 10.14302/issn.2326-0793.jpgr-12-207.
Surfactant protein A (SP-A) plays a number of roles in lung host defense and innate immunity. There are two human genes, and , and evidence indicates that the function of SP-A1 and SP-A2 proteins differ in several respects. To investigate the impact of SP-A1 and SP-A2 on the alveolar macrophage (AM) phenotype, we generated humanized transgenic (hTG) mice on the SP-A knockout (KO) background, each expressing human SP-A1 or SP-A2. Using two-dimensional difference gel electrophoresis (2D-DIGE) we studied the AM cellular proteome. We compared mouse lines expressing high levels of SPA1, high levels of SP-A2, low levels of SP-A1, and low levels of SP-A2, with wild type (WT) and SP-A KO mice. AM from mice expressing high levels of SP-A2 were the most similar to WT mice, particularly for proteins related to actin and the cytoskeleton, as well as proteins regulated by Nrf2. The expression patterns from mouse lines expressing higher levels of the transgenes were almost the inverse of one another - the most highly expressed proteins in SP-A2 exhibited the lowest levels in the SP-A1 mice and vice versa. The mouse lines where each expressed low levels of SP-A1 or SP-A2 transgene had very similar protein expression patterns suggesting that responses to low levels of SP-A are independent of SP-A genotype, whereas the responses to higher amounts of SP-A are genotype-dependent. Together these observations indicate that in vivo exposure to SP-A1 or SP-A2 differentially affects the proteomic expression of AMs, with SP-A2 being more similar to WT.
表面活性蛋白A（SP-A）在肺部宿主防御和固有免疫中发挥多种作用。人类有两个基因， 和 ，有证据表明SP-A1和SP-A2蛋白的功能在几个方面存在差异。为了研究SP-A1和SP-A2对肺泡巨噬细胞（AM）表型的影响，我们在SP-A基因敲除（KO）背景下培育了人源化转基因（hTG）小鼠，每只小鼠分别表达人SP-A1或SP-A2。我们使用二维差异凝胶电泳（2D-DIGE）研究了AM细胞蛋白质组。我们将表达高水平SP-A1、高水平SP-A2、低水平SP-A1和低水平SP-A2的小鼠品系与野生型（WT）和SP-A KO小鼠进行了比较。来自表达高水平SP-A2的小鼠的AM与WT小鼠最为相似，特别是与肌动蛋白和细胞骨架相关的蛋白质，以及受Nrf2调节的蛋白质。表达较高水平转基因的小鼠品系的表达模式几乎相反——SP-A2中表达最高的蛋白质在SP-A1小鼠中表达水平最低，反之亦然。每个表达低水平SP-A1或SP-A2转基因的小鼠品系具有非常相似的蛋白质表达模式，这表明对低水平SP-A的反应与SP-A基因型无关，而对较高水平SP-A的反应则依赖于基因型。这些观察结果共同表明，体内暴露于SP-A1或SP-A2会对AMs的蛋白质组表达产生不同影响，其中SP-A2与WT更为相似。
