The Diisopropylfluorophosphate Inhibitable Step in Antigen-Induced Histamine Release from Human Leukocytes

Abstract

DFP inhibits early events in antigen-induced histamine release from human leukocytes. If added to cells 5 min or more after antigen it is ineffective. If added with antigen it can be removed at 5 min but release will still be inhibited. In contrast, ethylenediaminetetraacetate (EDTA) and 2 deoxyglucose (2DG) still inhibit the reactions when added 5 min after antigen.

During incubation of leukocytes for 90 to 120 min at 0°C they react with specific antigen since they subsequently release significant quantities of histamine after washing and reincubation at 37°C without addition of antigen. Such priming at 0°C is at least equivalent to priming for 2 to 4 min at 37°C. During antigen priming at 0°C the cells are not activated beyond the step in the release sequence which is inhibited by diisopropylfluorophosphate (DFP). This is apparent from the undiminished inhibitory activity of DFP on these cells. Furthermore, cells primed with antigen at 0°C in the presence of DFP release as much histamine after washing and incubation at 37°C as control cells primed in the absence of DFP.

Incubation of leukocytes with specific antigen at 37°C for 3 min resulted in significant but not quite complete priming for subsequent histamine release in the absence of antigen. Most of these primed cells were not activated beyond the step inhibitable by DFP. However, some had completed the entire sequence including the release of histamine while others had not released their histamine but were not inhibited by DFP from subsequent release.

After 5 min incubation with antigen at 37°C almost all leukocytes had progressed beyond the stage which is inhibited by DFP.

Incubation of leukocytes at 37°C with DFP but without antigen for up to 15 min followed by washing did not impair subsequent antigen-induced histamine release by these cells. Thus, DFP was inhibitory under these conditions only after antigen activation of leukocytes.