I performed an ELISPOT for IgE using a biotinylated IgE-antibody and Streptavidin-ALP for detection of spots. I used bone marrow cells of two wild-type mice: one mouse was immunized with NP-CGG (T-cell dependent immunisation) and the other one was left unimmunised as a control.
When I analyzed the plate, I had IgE spots in case of both mice and the counts were pretty similar in both of them. Since this is bone marrow, i do not expect CSR (class switch recombination) or presence of IgE titers.
I read in a paper that incubating with an IgE antibody can stimulate the bone marrow mast cells to upregulate their Fc receptors and thus increasingly bind IgE on their surface. Is it possible that mast cells are binding some of the secreted IgE from my B-cells and thus generate false positives? Can anyone help with their reasoning or opinion on this problem?
Thanks a lot :)
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