Figure 5. GroEL activity on CD4+ T cells is not due to LPS contamination.

In order to test if the function came from possible LPS contamination, PMBCs were cultured with various antigens. (A) PBMCs were stimulated with rAaGroEL (20 µg/ml), LPS-removed-rAaGroEL (20 µg/ml; LPS was removed using Poly B supported resin). After 48 h stimulation cells were first labeled with CD4 antibodies. The cells, then, were fixed and permeabilized and incubated with anti-TNFα. Representative flow data shows TNFα expression among CD4+ T and non-CD4T cells. (B) PBMCs were incubated with rAaGroEL, Proteinase K-treated-rAaGroEL, heat-inactivated-Proteinase-K-treated-rAaGroEL and medium alone for 48 h and TNFα expression was measured. Representative flow data shows TNFα expression among CD4+ T and non-CD4T cells.