Detection of cytokine protein expression in mouse lung homogenates using suspension bead array.
BACKGROUND
The objective for this present study was to determine whether or not suspension bead array is a feasible method to detect changes in cytokine protein expression in mouse lung tissue homogenates. Here, we report on suspension bead array as a feasible method for detection of lipopolysaccharide (LPS)-evoked changes in cytokine protein expression in mouse lung tissue homogenates.


METHODS
Mice were treated (0.2 ml, intraperitoneal, i.p.) with phosphate buffered saline (PBS) or LPS (0.25 mg/ml) and sacrificed at either 2- or 24-hours post treatment. Formalin-fixed and paraffin-embedded lung sections were evaluated by light microscopy. Flash frozen lung tissues were homogenized for measurement of various cytokine protein expression levels using suspension bead array, antibody array and ELISA. Comparison between groups was performed using a Wilcoxon signed rank test.


RESULTS
Pulmonary perivascular edema and an accumulation of mixed cell infiltrates within blood and lymphatic vessels, as well as in the adjacent interstitium, were present at both 2- and 24-hours following LPS treatment. A minimal increase in the number of alveolar macrophages was also observed in the 24-hour LPS-treated mice only. The suspension bead array assay revealed statistically significant increases in mouse lung tissue homogenate levels of interleukin-6 (IL-6) and granulocyte/macrophage colony-stimulating factor (GM-CSF) proteins and a decrease in IL-2 protein at 24-hours post LPS-treatment only. Similar cytokine protein expression patterns were observed using antibody array. Significantly increased IL-6 protein expression levels were also detected using ELISA, which correlated with the suspension bead array data.


CONCLUSIONS
The present study shows that suspension bead array is a feasible method to detect changes in cytokine protein expression in mouse lung tissue homogenates.
McDuffie
Eric
E
Drug Safety Evaluation Department, Esperion, a Division of Pfizer Research and Development, Ann Arbor, Michigan, 48108 USA.
Obert
Leslie
L
Worldwide Safety Sciences Department, Pathology Section, Pfizer Research and Development, Ann Arbor, Michigan, 48105 USA.
Chupka
Jonathan
J
Drug Safety Evaluation Department, Esperion, a Division of Pfizer Research and Development, Ann Arbor, Michigan, 48108 USA.
Sigler
Robert
R
Drug Safety Evaluation Department, Esperion, a Division of Pfizer Research and Development, Ann Arbor, Michigan, 48108 USA.