Identification of MIC 11 antigen as an epitope of the CD59 molecule.
The MIC 11 antigen is expressed on human cells and is characterized by reaction with a monoclonal antibody (mAb), 16.3A5. The gene controlling MIC 11 was recently mapped to the p13 region of chromosome 11 within 500 kb of the gene encoding CD59, a complement regulatory protein. The present report investigates the antigenic relationship between these cell-membrane determinants and sets out evidence that MIC 11 and CD59 are encoded by the same gene. Western blotting of human erythrocyte membrane proteins and purified membrane CD59 showed that 16.3A5 anti-MIC 11 antibody bound to a 19-24,000 MW band with the characteristic appearance of CD59 protein, and gave staining patterns identical to those obtained with the CD59 antibody, BRIC 229. The binding of 16.3A5 monoclonal IgG to purified urine-derived CD59 in enzyme-linked immunosorbent assay (ELISA) was inhibited by YTH 53.1 rat CD59 antibody, indicating that the MIC 11 epitope is the same as, or close to, that recognized by CD59 antibodies such as YTH 53.1, BRIC 229 and 2/24. Prior exposure of erythrocytes to 16.3A5 anti-MIC 11 also reduced the ability of the CD59 antibodies, BRIC 229 and YTH 53.1, to block the complement-inhibiting function of membrane CD59. Anti-MIC 11 antibody alone, however, had no inhibitory effect on CD59 function. This may be due to its relatively low binding affinity or to some slight difference in epitope specificity. Further studies using immunofluorescence showed that the MIC 11 epitope, like CD59, is absent from EBV-B cells lacking GPI-anchored proteins and from a B-cell line specifically deficient in CD59 protein. Overall, the results provide strong evidence that MIC 11 is a determinant on the CD59 molecule.
Davies
A
A
Molecular Immunopathology Unit, Medical Research Council Centre, Cambridge, UK.
Wilson
A B
AB
Bramley
J C
JC
Willers
C
C
Van Heyningen
V
V
Bickmore
W A
WA
Lachmann
P J
PJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D015703:Antigens, CD / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D001667:Binding, Competitive; D015153:Blotting, Western; D018957:CD59 Antigens; D002461:Cell Line, Transformed; D003169:Complement Inactivator Proteins / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D004910:Erythrocyte Membrane / Q000276:immunology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D004854:Herpesvirus 4, Human; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018957:CD59 Antigens; D003169:Complement Inactivator Proteins; D000939:Epitopes; D008562:Membrane Glycoproteins