Serologic subtyping of HLA-DR8 by means of the cytotoxic human monoclonal antibody 5643.
EBV-transformed B cells from a multiparous woman with anti-HLA antibodies were hybridized with CB-F7 heteromyeloma cells. The resulting hybridoma produced the cytotoxic human IgM(kappa) mAb 5643. Testing of the hybridoma supernatant against HLA homozygous cell lines demonstrated positive reactions (titer range 128-512) only with three cell lines that carried the DR (alpha, beta 1*0801) subtype of DR8. There was no reaction with cells expressing other variants of DR8; i.e., carrying the DR beta 1*0802, 0803, or 0804 chains. The only difference between the DR beta 1*0801 and DR beta 1*0802/0804 chains and between the DR beta 1*0801 and 0803 chains is a single aa substitution, a Ser-->Asp substitution at residue 57 and a Phe-->Ile substitution at residue 67, respectively. Residues 57 and 67 are both situated on the alpha-helix of the DR beta chain. The distance between residues 57 and 67 is two full turns of the alpha-helix; i.e., about 1 nm. It is possible that both 57 Ser and 67 Phe are directly involved in the epitope recognition by mAb 5643.
Viken
H D
HD
Institute of Transplantation Immunology, National Hospital, Oslo, Norway.
Thoresen
A B
AB
Knutsen
I
I
Thorsby
E
E
Hansen
T
T
D000483:Alleles; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001171:Arthritis, Juvenile / Q000175:diagnosis / Q000235:genetics / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D006681:HLA-D Antigens / Q000032:analysis / Q000235:genetics; D006684:HLA-DR Antigens / Q000032:analysis* / Q000235:genetics; D059810:HLA-DR Serological Subtypes; D006650:Histocompatibility Testing; D006801:Humans; D006825:Hybridomas; D007075:Immunoglobulin M / Q000097:blood; D008969:Molecular Sequence Data; D009664:Norway; D014407:Tumor Cells, Cultured; D044465:White People / Q000235:genetics
D000911:Antibodies, Monoclonal; D006681:HLA-D Antigens; D006684:HLA-DR Antigens; D059810:HLA-DR Serological Subtypes; C041559:HLA-DR8 antigen; D007075:Immunoglobulin M