Research objective. The primary objective of this research was to test the hypothesis that administering rGM-CSF to neonatal LPL−/− pups (in the temporal window of AM development) would protect LPL−/− mice during subsequent bacterial lung infection. This objective was defined before initiation of the experiments and data analysis.

Experimental design. The experimental design was a controlled laboratory experiment using genetically modified mice.

Randomization. Pups within each litter of mice were randomly assigned to receive intranasal rGM-CSF or PBS (control). In one set of experiments, pups within litters were randomized to receive subcutaneous rGM-CSF or PBS (control).

Blinding. Histology sections were read by a veterinary pathologist who was not associated with the study outcomes and was blinded to mouse genotype and treatment group (slides were provided with coded labels). Because of the technical challenges associated with working with timed pregnancies and neonatal pups, and the necessity of ensuring that pups receiving PBS could be consistently distinguished from the pups receiving rGM-CSF (pups were marked with permanent ink, but this would wear off if not reapplied), most of the studies could not be blinded. However, the outcomes are all objectively quantifiable [flow cytometry, colony forming units (CFU), etc.].

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