title: A method of generating antibodies against exogenously administered self-antigen by manipulating CD4+CD25+ regulatory T cells.
abstract: The generation of antibodies against self-antigens or antigens having a high degree of structural homology with self-antigens is a difficult task because of immunological tolerance. CD4(+)CD25(+) regulatory T cells play an important role in maintaining peripheral tolerance. Sakaguchi et al. previously reported that the transfusion of CD25(+) cell-depleted mouse splenocytes into syngeneic nude mice results in a breaking of peripheral tolerance that leads to the development of autoimmunity. In this study, we attempted to apply this mouse model to the generation of antibodies against self-antigens. We depleted CD25(+) cells from BALB/c mouse splenocytes and transferred the rest of the cells into syngeneic nude mice. The animals were immunized with mouse thyroglobulin. We observed a significant increase of the anti-mouse thyroglobulin antibody titer in the group of mice immunized twice within 10 days after the cell transfer (P<0.05). From these mice, we established hybridoma cell lines producing anti-mouse thyroglobulin monoclonal antibodies, including a clone with a dissociation constant of 10(-8)M. Control nude mice which received CD25(+) cell-containing BALB/c splenocytes did not produce anti-mouse thyroglobulin antibodies. When the CD25(-)cell-transferred nude mice were immunized with mouse Gα12, another self-antigen, anti-Gα12 antibodies were produced in the sera. This method should prove highly useful in the generation of antibodies against self-antigens or antigens for which the structure is highly conserved across species.
authors.affiliation: Department of Molecular Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8904, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Iwanari
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Nakada-Nakura
authors.forename: Osamu
authors.initials: O
authors.lastname: Kusano-Arai
authors.forename: Nobuchika
authors.initials: N
authors.lastname: Suzuki
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kodama
authors.forename: Toshiko
authors.initials: T
authors.lastname: Sakihama
authors.forename: Takao
authors.initials: T
authors.lastname: Hamakubo
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000917:Antibody Formation; D001324:Autoantigens / Q000008:administration & dosage / Q000276:immunology*; D002469:Cell Separation; D005260:Female; D007114:Immunization / Q000379:methods*; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D013154:Spleen / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
chemical_list: D000906:Antibodies; D001324:Autoantigens; D053645:Interleukin-2 Receptor alpha Subunit