Construction of novel class I histocompatibility antigens by interspecies exon shuffling.
Human and mouse class I histocompatibility antigens share considerable structural homology at both the protein and DNA sequence level. This homology has allowed the production of hybrid class I molecules by the reciprocal exchange of DNA sequences corresponding to equivalent domains of HLA-B7 and either H-2Ld or H-2Dd. It is shown that these genes give rise to protein products that are stably expressed on the surface of murine L cells after DNA-mediated gene transfer. These proteins express only those monoclonal antibody-defined H-2 determinants that are expected based on their genetic construction. The molecules have allowed the localization of a number of polymorphic and monomorphic HLA-specific epitopes. In all but one case, expression of an epitope on a domain does not appear to be influenced by the replacement of adjacent human domains with their murine equivalents, suggesting a considerable degree of structural independence of the domains. Cells expressing the hybrid molecules have also been tested as targets for a panel of HLA-B7-specific cytotoxic T cell clones. The results show that the polymorphic determinants recognized by these clones map to the alpha 1 and alpha 2 domains of the HLA-B7 molecule. No evidence for an influence of species-related amino acid sequence differences in the third extracellular domain on T cell recognition was seen. The results are discussed in light of the proposed domain structure of the class I proteins and the potential use of such molecules for further functional studies.
Engelhard
V H
VH
Yannelli
J R
JR
Evans
G A
GA
Walk
S F
SF
Holterman
M J
MJ
D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D006183:H-2 Antigens / Q000235:genetics* / Q000276:immunology; D006680:HLA Antigens / Q000235:genetics* / Q000276:immunology; D015793:HLA-B7 Antigen; D064532:Histocompatibility Antigen H-2D; D006801:Humans; D006822:Hybrid Cells / Q000276:immunology; D007739:L Cells / Q000276:immunology; D051379:Mice; D014176:Protein Biosynthesis; D013045:Species Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D000911:Antibodies, Monoclonal; D006183:H-2 Antigens; D006680:HLA Antigens; D015793:HLA-B7 Antigen; D064532:Histocompatibility Antigen H-2D; D004262:DNA Restriction Enzymes