Expression and Immunogenicity of the Mycobacterial Ag85B/ESAT-6 Antigens Produced in Transgenic Plants
by Elastin-Like Peptide Fusion Strategy
Impact of purified ELP on mouse and bovine immune cells. (a) ELP effect on cell proliferation in mouse spleen cells and bovine PBMC. CFSE-stained mouse spleen cells and bovine PBMC were stimulated for 4 days with 5 g/mL of purified ELP (ELP) or mitogen ConA (ConA) or left untreated (Control). Cells were then collected and processed by flow cytometry for measurement of proliferating cells. The histograms show the FL1 emission values (FL1-H) for one representative experiment for each animal cell type out of two duplicate experiments. Results are expressed in percentage of cells that had undergone cell divisions, that is, with decreased CFSE fluorescence intensity. The control histogram (unstimulated cells) was filled in grey; the doted black line corresponded to ELP and the grey solid line to ConA stimulation. (b) ELP effect on maturation marker expression in mouse bone marrow-derived dendritic cells (mBM-DCs). Mouse BM-DCs ( cells/well) were stimulated for 24 hours with purified ELP (2.5 and 5 g/mL), LPS (control +) or left untreated (control −). Cells were immunostained of the following cell surface markers: CMH II, CD40, CD80, CD86 and processed by flow cytometry as described in Material and Methods. The histogram presents the mean ( SD) of the percentages of fluorescing cells for each cell surface marker for two experiments done in duplicate.