Mechanisms of asthma and allergic inflammationRelationship between spleen tyrosine kinase and phosphatidylinositol 5′ phosphatase expression and secretion from human basophils in the general population
Section snippets
Materials and buffers
The Online Repository (www.jacionline.org) provides the description of standard materials, buffers, and antibodies used for these studies. Briefly, various piperazine-N,N′-bis(2-ethanesulfonic acid) buffers, as described previously, were used for short periods of stimulation, and supplemented RPMI-1640 was used for overnight incubations.17 RPMI-1640 for overnight cultures contained 8 μg/mL gentamycin and 300 μg/mL human serum albumin, supplemented to 1 mmol/L final Ca++ concentration. The syk
Results
To assess the variation in interassay sample error, whole cell lysates from 9 different subjects were run in multiple Western blots. From these measurements, we found approximately ± 24% (error relative to measured amount) variation for the syk kinase determination and ± 11% variation for SHIP1. We also examined expression levels in basophils of 9 subjects with a second purification and Western analysis; expression levels showed a relative variation of 21% and 35% for syk and SHIP1,
Discussion
Previous studies on the relationship between signal element expression and basophil function examined extreme differences in function. In addition, the codependent relationship between signal element expression and function has not been examined. The current studies provide a broader context for signal elements expression. There are certainly many signaling elements absolutely required for mediator release, but it appears that syk expression is a focus of natural variation. Indeed, the
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Supported by National Institutes of Health grant AI20253.
Disclosure of potential conflict of interest: The author has declared that he has no conflict of interest.