I am looking for an alternative way of doing basophil activation test(BAT) using passive sensitization. Technically, sensitizing the besophils from the healthy individuals' by patients sera and evaluating the up regulation of CD 63 by BAT, Can any one tell me is it possible?
You would need to remove the IgE that is already bound to fcERI. As this is a very high affinity receptor most receptors will already be bound by IgE from your healthy individual. I think you can strip wih mild acid solution.
alternatively, why dont you use a cell line such as RBL to sensitize with serum and test degranulation?
Many thanks for your answer and the valid suggestion Jolien, would the cell line behave like normal human basophil? Do we have to add cytokines to prime the cell line? Is it okay to use a RBL instead of a human basophil cell line?
Behavior like a normal basophil: Probably not exactly as it is a tumor cell line. However, if you are just interested into whether your serum leads to degranulation through FcERI then it will probably work just fine. It is an adherent cell line so usually people check degranulation using the beta-hexosaminidase assay rather then CD63 for flow.
You don't need to prime the cells with cytokine, but you will need to use RBL cellline transfected with human FcERI so that it works with humans serum..
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