Expression of rabbit interleukin-4 and characterization of its biologic activity on T and B-cells.
The purpose of the current study was to express recombinant rabbit IL-4 (rRbIL-4) and to characterize its biological activity. The cDNA of RbIL-4 was cloned into an insect cell expression vector that allowed for constitutive expression in Sf9 cells and incorporated a 6-histidine tag on the recombinant protein for purification. The purified protein corresponded to the predicted size of rRbIL-4 and was recognized by an anti-human IL-4 antibody in immunoblotting. As shown for IL-4 from other species, a dose-dependent proliferative response was observed in T-lymphoblasts cultured with rRbIL-4. rRbIL-4 also induced increased expression of MHC class II molecules on the surface of rabbit B-cells in a dose-dependent manner. These results indicate that we have produced recombinant rabbit IL-4 that exhibits expected biological activity on rabbit B and T-cells.
Leader
Brandon T
BT
Department of Pathobiology, University of Washington, Seattle, WA 98195, USA. btleader@u.washington.edu
VanVoorhis
Wesley C
WC
Lukehart
Sheila A
SA
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D015153:Blotting, Western / Q000662:veterinary; D005434:Flow Cytometry; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D015847:Interleukin-4 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D008213:Lymphocyte Activation; D008297:Male; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011817:Rabbits / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D018411:Spodoptera / Q000235:genetics; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D000949:Histocompatibility Antigens Class II; D011994:Recombinant Proteins; D015847:Interleukin-4; D012313:RNA