We just tried using Pharmingen's PE-conjugated anti-human IL4 (clone 8D4-8, recommended for nonhuman primates in the Fall 2001 issue of "Hotlines") with freshly isolated rhesus peripheral blood mononuclear cells that were cultured overnight in the presence of Brefeldin A and the in the presence or absence of Staph enterotoxin B (SEB). Using 0.2 ug of antibody (Pharmingen recommends < 0.25 ug per 1 million cells) we observed equivalent staining of subpopulations of unstimulated and stimulated T cells (intracellular CD3+), and also observed equivalent staining of a subpopulation of CD3-negative lymphocytes (non T cells). In contrast, when we used PE-conjugated anti-IFNgamma or anti-IL2 in the same experiment we observed staining only of SEB-stimulated T cells, as expected. Does anyone have any comments on why we are not seeing T cell-specific, stimulation dependent IL4 staining? Gib Otten Chiron Corp.
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