title: Production and characterization of monoclonal antibodies against porcine interleukin-4.
abstract: Interleukin 4 (IL-4) is an important regulatory cytokine produced by activated T lymphocytes and mast cells, and regulates the growth and differentiation of cells such as B and T lymphocytes. In the present study, recombinant thioredoxin (Trx)-porcine IL-4 (pIL-4) fusion protein was prepared by Escherichia coli (E. coli), and by using this protein as an immunogen, monoclonal antibodies (mAbs) against pIL-4 were produced to establish a basis for a research on immune responses in pigs. Six stable hybridoma cell lines were successfully established and specific binding of each mAb to recombinant pIL-4 produced by E. coli and insect cells infected with recombinant baculovirus was shown by enzyme-linked immunosorbent assay (ELISA) and/or immunoblot analysis. Isotype analyses of these mAbs revealed that the subclass of 5 out of 6 mAbs was IgG1 and the rest was IgG2b. Further, assessment of their epitopes by competition binding assay indicated that the mAbs obtained in this study bound to 4 different epitopes. The recombinant proteins and mAbs produced in this study will be useful tools for the assessment of porcine immune system.
authors.affiliation: Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Yoji
authors.initials: Y
authors.lastname: Ishikura
authors.forename: Hirotomo
authors.initials: H
authors.lastname: Kato
authors.forename: Tomomi
authors.initials: T
authors.lastname: Hashimoto
authors.forename: Yorihiro
authors.initials: Y
authors.lastname: Nishimura
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Iwata
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D000939:Epitopes; D004926:Escherichia coli; D006825:Hybridomas; D015847:Interleukin-4 / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D013552:Swine
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D015847:Interleukin-4