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1 Cellular and Molecular Physiology, Penn State University College of Medicine, Hershey, Pennsylvania, United States
2 Dept. of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania, United States
3 Pediatrics, Pennsylvania State University College of Medicine, Hershey, Pennsylvania, United States
4 Penn State University College of Medicine, Hershey, Pennsylvania, United States
5 Pediatrics, Penn State University College of Medicine, Hershey, Pennsylvania, United States
6 Division of Pulmonary and Critical Care Medicine, East Carolina Univerity Brody School of Medicine, Greenville, North Carolina, United States
7 Pulmonary Critical Care Med., Cleveland Clinic Foundation, Cleveland, Ohio, United States
8 Pediatrics, The Penn State University, Hershey, Pennsylvania, United States
* To whom correspondence should be addressed. E-mail: jfloros{at}psu.edu.
The human surfactant protein A (SP-A) locus consists of two functional genes (SP-A1, SP-A2) with gene specific products exhibiting qualitative and quantitative differences. The aim here was two-fold: 1) generate SP-A1 gene-specific antibody, and 2) use this to assess gene-specific SP-A content in the bronchoalveolar lavage fluid (BALF). An SP-A1 specific polyclonal antibody (hSP-A1_Ab68-88_Col) was raised in chicken and its specificity determined by immunoblot and ELISA using mammalian CHO cell-expressed SP-A1 and SP-A2 variants, and by immunofluorescence with stably transfected CHO cell lines expressing SP-A1 or SP-A2 variants. SP-A1 content was evaluated according to age and lung status. A gradual decrease (p<0.05) in SP-A1/SP-A ratio was observed in healthy subjects (HS) with increased age, although no significant change was observed in total SP-A content among age groups. Total SP-A and SP-A1 content differed significantly between alveolar proteinosis (AP) patients and HS, with no significant difference observed in SP-A1/SP-A ratio between AP and HS. The cystic fibrosis (CF) ratio was significantly higher compared to AP, HS, and non-cystic fibrosis (NCF), even though SP-A1 and total SP-A were decreased in CF compared to most of the other groups. The ratio was higher in culture positive versus culture negative samples from CF and NCF (p=0.031). A trend of an increased ratio was observed in culture positive CF (0.590±0.10) compared to culture positive NCF (0.3680±0.085). In summary, we developed and characterized an SP-A1 gene specific Ab and used it to identify gene-specific SP-A content in BALFs as a function of age and lung health.
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