Expression of a functional chimeric Ig-MHC class II protein.
We have generated a chimeric protein molecule composed of the alpha- and beta-chains of the MHC class II I-E molecule fused to antibody V regions derived from anti-human CD4 mAb MT310. Expression vectors were constructed containing the functional, rearranged gene segments coding for the V region domains of the antibody H and L chains in place of the first domains of the complete structural genes of the I-E alpha- and beta-chains, respectively. Cells transfected with both hybrid genes expressed a stable protein product on the cell surface. The chimeric molecule exhibited the idiotype of the antibody MT310 as shown by binding to the anti-idiotypic mAb 20-46. A protein of the anticipated molecular mass was immunoprecipitated with anti-mouse IgG antiserum. Furthermore, human soluble CD4 did bind to the transfected cell line, demonstrating that the chimeric protein possessed the binding capacity of the original mAb. Thus, the hybrid molecule retained: 1) the properties of a MHC class II protein with regard to correct chain assembly and transport to the cell surface; as well as 2) the Ag binding capacity of the antibody genes used. The generation of hybrid MHC class II molecules with highly specific, non-MHC-restricted binding capacities will be useful for studying MHC class II-mediated effector functions such as selection of the T cell repertoire in thymus of transgenic mice.
Zwirner
J
J
Department of Immunology, Scripps Research Institute, La Jolla, CA 92037.
Weissenhorn
W
W
Karlsson
L
L
Becker
A
A
Rieber
E P
EP
Riethmüller
G
G
Weiss
E H
EH
Peterson
P A
PA
Widera
G
G
D000818:Animals; D001666:Binding Sites, Antibody; D015704:CD4 Antigens / Q000276:immunology; D002462:Cell Membrane / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D005803:Genes, Immunoglobulin; D005802:Genes, MHC Class II; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000378:metabolism; D066298:In Vitro Techniques; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D015704:CD4 Antigens; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins