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Search for Signs of Inflammatory Disease

See Related Article: Curing Cancer in Mice

You fall on your shoulder and tear some cartilage, causing bone to rub against bone. Your shoulder becomes inflamed and begins to hurt because cytokine, a small signal protein secreted by your immune system, has recruited white blood cells to clean up the damage.

ORNL biologist Steve Kennel recently worked on a project to measure levels of a specific cytokine in samples from mice. The goal was to screen for mice that are likely to develop inflammatory diseases similar to those in humans, such as arthritis, psoriasis, and inflammatory bowel disease.

Greg Hurst checks a sample plate with 100 wekks containing various protein fragments
Greg Hurst checks a sample plate with 100 wells containing various protein fragments. The plate will be inserted into the time-of-flight mass spectrom-eter shown at right. This instrument is used to identify and characterize sample proteins. (Photo by Curtis Boles, enhanced by Gail Sweeden.)

In a study supported by the Laboratory Directed Research and Development Program at ORNL, Kennel collaborated with Greg Hurst, a mass spectrometry expert in ORNL's Chemical and Analytical Sciences Division. They developed a technique that combines affinity chromatography with mass spectrometry to separate three specific cytokines out of the 100 or so different cytokines in mouse blood or fluid extracted from mouse cells.

"I obtained antibodies specific to different cytokines and attached them to beads," Kennel says. "These beads were tossed into a soup of proteins to fish out specific cytokines for analysis by matrix-assisted laser desorption ionization mass spectrometry.

"We were able to identify each cytokine. But because we had only small amounts of blood from a mouse, we were unable to detect unusually high levels of our target protein-the tumor necrosis factor alpha cytokine. So, we could not be sure these test mice were showing an inflammation response."

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