title: Isotype modulation of idiotypic expression in recombinant isotypic variants of MOPC 315.
abstract: The influence of the CH1 domains of various isotypes on the expression of four Id of the IgA 2 mouse myeloma protein MOPC 315 was assessed. To this end, mammalian expression vectors containing the rearranged MOPC 315 VH gene along with the H chain genes of various isotypes were constructed. These vectors were then transfected into the L chain-expressing MOPC 315.26 cell line to produce the rIg. The effect of polyvalency on the ability of Ig to bind anti-idiotypic antibodies was tested by comparing idiotypic expression in a competitive ELISA using reduced and nonreduced MOPC 315 IgA and IgM species. Reduction produced a two- to fivefold decrease in their ability to inhibit the binding of three anti-idiotypic antibodies, but not that of the functionally univalent antibody D10. In contrast, reduction of MOPC 315 IgG proteins did not affect the binding of the anti-Id mAb, indicating that reduction of the interchain disulfide bonds did not alter idiotypic expression. The expression of idiotopes on reduced mouse rIgA, IgM and IgG and human IgG MOPC 315 molecules was then compared. The results showed that both human and mouse IgG recombinant antibodies exhibited an enhanced expression of the idiotopes recognized by antibodies D10 and F1, as compared to MOPC 315 IgA and IgM molecules. In contrast, the expression of idiotopes recognized by A2 and G3 mAb was not influenced by the H chain isotype. These data support the hypothesis that the conformation of certain idiotopes is modulated by the isotype of the CH1 domain.
authors.affiliation: Department of Immunology, University of Toronto, Ontario, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Rinfret
authors.forename: C
authors.initials: C
authors.lastname: Horne
authors.forename: H
authors.initials: H
authors.lastname: Boux
authors.forename: A
authors.initials: A
authors.lastname: Marks
authors.forename: K J
authors.initials: KJ
authors.lastname: Dorrington
authors.forename: M
authors.initials: M
authors.lastname: Klein
mesh_terms: D000478:Alkylation; D000818:Animals; D007070:Immunoglobulin A / Q000032:analysis*; D007130:Immunoglobulin Idiotypes / Q000032:analysis*; D007132:Immunoglobulin Isotypes / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis; D051379:Mice; D009194:Myeloma Proteins / Q000276:immunology*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D007070:Immunoglobulin A; D007130:Immunoglobulin Idiotypes; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D009194:Myeloma Proteins; D011994:Recombinant Proteins