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Undergrad Helps Find Possible West Nile Cure
by Doug Main
"At first, it was like a whole new world to me," says Christopher Doane, 21, of his work in Michael S. Diamond's lab.
Awarded an HHMI undergraduate fellowship after his freshman year at Washington University in St. Louis, Doane was assigned to generate monoclonal antibodies against a specific protein of the West Nile virus (WNV).
The goal sounded difficult, and even veteran researchers attest to the complexity of the process. Diamond, who researches infectious diseases at Washington University School of Medicine, taught Doane how to create special antibody-producing cells, called hybridomas, to combat WNV—a mosquito-borne pathogen that can afflict humans with anything from mild flu symptoms to brain swelling and occasionally even death.
Of his great grandfather, who helped found HHMI, Christopher Doane says, "I am just amazed a person could help so many people in his lifetime."
The laboratory process involves fusing tumor cells with the spleen cells of lab mice previously inoculated with WNV's E protein. If the fusion is done just right, the two cells create a hybridoma and secrete monoclonal antibodies capable of binding to the E protein, thereby neutralizing the virus. To increase the odds of success, a number of fusions are attempted at once in a multiwell plate; bright fluorescence within a well signals a successful binding.
During his first 5 months, Doane achieved little success. He kept at it into the next school year. Then, one day near Thanksgiving, his luck changed.
"The whole well plate lit up," says Doane. Preliminary results suggested he had generated 30 different hydridomas, each secreting antibody against WNV E protein. Doing further screening, Doane and his colleagues observed that one antibody in particular, named E16, had surprising binding ability--it neutralized 10 different strains of WNV, preventing the virus from infecting cells. Further testing showed that E16 prevented infected mice from dying, even when administered up to 5 days after exposure.
Photo: Bob Hower / Quadrant Photography