Zsengellér Z K, Reed J A, Bachurski C J, LeVine A M, Forry-Schaudies S, Hirsch R, Whitsett J A
Division of Pulmonary Biology, Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Hum Gene Ther. 1998 Sep 20;9(14):2101-9. doi: 10.1089/hum.1998.9.14-2101.
Mutation of the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene by homologous recombination causes progressive pulmonary alveolar proteinosis (PAP) in GM-CSF-deficient mice (GM-/-). The present study tested whether adenovirus-mediated expression of GM-CSF alters the progression of PAP in GM-/- mice. Adult mice were pretreated with an anti-T cell receptor (TCR) antibody to block T cell-mediated immune response, followed by intratracheal instillation of deltaE1-E3 replication-deficient adenovirus expressing mouse GM-CSF (Av1mGM). Mice were killed 1, 3, and 5 weeks after treatment to assess lungs for GM-CSF, surfactant protein B (SP-B), alveolar macrophage maturation, and type II cell proliferation. GM-CSF was detected in BAL fluid from GM-/- mice 1 week after Av1mGM treatment, and GM-CSF mRNA was detected by RT-PCR through 5 weeks. Five weeks after Av1mGM treatment, PAP was improved and SP-B decreased as assessed by ELISA and immunostaining. Increased numbers of alveolar macrophages stained with alpha-naphthyl acetate esterase (alpha-NAE) following treatment with Av1mGM. Local expression of GM-CSF with a recombinant adenovirus ameliorated PAP in the GM-/- mice in association with enhanced maturation of alveolar macrophages.
通过同源重组使粒细胞巨噬细胞集落刺激因子（GM-CSF）基因突变会导致GM-CSF缺陷小鼠（GM-/-）出现进行性肺泡蛋白沉积症（PAP）。本研究检测了腺病毒介导的GM-CSF表达是否会改变GM-/-小鼠PAP的病程。成年小鼠先用抗T细胞受体（TCR）抗体进行预处理以阻断T细胞介导的免疫反应，然后经气管内滴注表达小鼠GM-CSF的缺失E1-E3区的复制缺陷型腺病毒（Av1mGM）。在治疗后1、3和5周处死小鼠，评估肺组织中的GM-CSF、表面活性蛋白B（SP-B）、肺泡巨噬细胞成熟度和II型细胞增殖情况。Av1mGM治疗1周后在GM-/-小鼠的支气管肺泡灌洗（BAL）液中检测到GM-CSF，通过逆转录聚合酶链反应（RT-PCR）在5周内均可检测到GM-CSF mRNA。Av1mGM治疗5周后，通过酶联免疫吸附测定（ELISA）和免疫染色评估发现PAP有所改善，SP-B减少。用Av1mGM治疗后，用α-萘乙酸酯酶（α-NAE）染色的肺泡巨噬细胞数量增加。用重组腺病毒在局部表达GM-CSF可改善GM-/-小鼠的PAP，同时伴有肺泡巨噬细胞成熟度增强。
