title: Antibody production by injection of living cells expressing non self antigens as cell surface type II transmembrane fusion protein.
abstract: Antigen expression and purification are laborious, time consuming and frequently difficult steps in the process of antibody production. In the present study, we developed a method avoiding these two steps. This method relies on the injection of histocompatible living cells stably expressing the antigen as a cell surface type II transmembrane fusion protein. A vector, nicknamed pCD1-CD134L, was constructed to express the antigen fused at the carboxyterminal end of the human CD134 ligand (CD134L) type II transmembrane protein on the surface of eucaryotic cells. This vector was shown to induce cell surface expression of epitopes from human c-Myc (soluble protein), uterogloblin-related protein 1 (secreted protein) and CD94 (type II transmembrane protein). Using this vector, we developed a method to produce antibodies without antigen production. The flowchart of this method is as follows: (i) cloning of the antigen in the pCD1-CD134L vector; (ii) production of a histocompatible cell line stably expressing the CD134L-antigen fusion protein; (iii) testing for cell surface expression of the fusion protein by targeting the CD134L carrier; and (iv) prime-boost immunisation with living cells expressing the fusion protein. This method was successfully used for production of polyclonal antibodies raised against Ixodes ricinus calreticulin (secreted protein) in mice and for production of monoclonal antibodies raised against an epitope of Vaccinia virus A56 (type I transmembrane protein) protein in rat. The present study is the first to demonstrate the use of a type II transmembrane protein as a carrier for cell surface display of antigens.
authors.affiliation: Experimental Immunology Unit, Université Catholique de Louvain, Brussels, Belgium. yannick.nizet@uclouvain.be
authors.forename: Yannick
authors.initials: Y
authors.lastname: Nizet
authors.forename: Laurent
authors.initials: L
authors.lastname: Gillet
authors.forename: Hélène
authors.initials: H
authors.lastname: Schroeder
authors.forename: Corinne
authors.initials: C
authors.lastname: Lecuivre
authors.forename: J
authors.initials: J
authors.lastname: Louahed
authors.forename: J-C
authors.initials: JC
authors.lastname: Renauld
authors.forename: Pierre
authors.initials: P
authors.lastname: Gianello
authors.forename: Alain
authors.initials: A
authors.lastname: Vanderplasschen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D005260:Female; D018884:Ixodes; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D053278:OX40 Ligand / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D053278:OX40 Ligand; D011993:Recombinant Fusion Proteins; C089251:TNFSF4 protein, human