Figure 1. APC-T Cell Partners Upregulate Complement mRNAs and the RNAs Produce Proteins.
(A) OT-II T cells were incubated for 1 hr with WT DCs ± 0.1 μM OVA323–339 and flow separated (with anti-CD3 and anti-CD11c,) and complement mRNA expression in each partner was measured by qPCR.
(B) OT-II cells and DCs were flow separated at increasing times, and complement IL-2, IFN-γ, IL-12, and IL-23 gene expression was measured by qPCR.
(C) The left side shows representative (rep) histograms (four exps; linear scales) depicting C5aR or C3aR on OT-II cells and DCs before (no OVA) and after 1 hr interaction with OVA. The right side shows that after 24 hr of interaction of DCs with OT-II cells ± OVA, flow-separated cells were cultured for 4 hr, and supernatants were blotted for C3a and C5a; stds = 2 ng.
(D) Kinetics of C5aR, C3aR, and DAF protein expression on OT-II T cells and DCs during interaction with ova. Fold increase is relative to no OVA cultures. DAF levels on the DCs were low at all time points.
(E) After interaction of OT-II cells with DCs ± ova for 18 hr with 4 μg/ml anti-B7.1 and anti-B7.2 mAbs or control IgG, mRNAs in flow-separated cells were assayed for C3, C3aR, C5aR, and IFNγ gene expression by qPCR. In parallel cultures, IFNγ was assessed by ELISPOT. No complement or cytokine upregulation occurred without T cells. Data are normalized to no OVA. Each experiment is representative of two to four replicate studies. *p < 0.05 versus controls. All error bars are ± SD.