Cloning the antibody response in humans with inflammatory CNS disease: isolation of measles virus-specific antibodies from phage display libraries of a subacute sclerosing panencephalitis brain.
We have developed a strategy to identify the disease-relevant antigens in a chronic inflammatory CNS disease exhibiting intrathecally expressed oligoclonal IgG. Using subacute sclerosing panencephalitis (SSPE), a chronic inflammatory measles virus infection of the brain as a model system, we constructed a phage display antibody Fab library from the amplified products of IgG expressed in the brain. Selection of the library against measles virus-infected cell lysates yielded four distinct Fabs which, by ELISA and by immunostaining, reacted specifically with measles virus-infected cells. Three Fabs immunoprecipitated a 72 kDa protein from infected cell cultures corresponding to the measles virus phosphoprotein. The fourth Fab immunoprecipitated and recognized by immunoblotting a 60 kDa protein corresponding to the measles virus nucleoprotein. The results demonstrate that functional antibodies from an inflammatory CNS disease can be expressed in bacteria and used to identify disease-relevant antigens. This approach could be applied to chronic inflammatory CNS diseases of unknown cause such as multiple sclerosis.
Burgoon
M P
MP
Department of Neurology, University of Colorado Health Sciences Center, Denver 80262, USA.
Williamson
R A
RA
Owens
G P
GP
Ghausi
O
O
Bastidas
R B
RB
Burton
D R
DR
Gilden
D H
DH
D000293:Adolescent; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000032:analysis / Q000302:isolation & purification; D001435:Bacteriophages; D001921:Brain / Q000276:immunology / Q000821:virology; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D015723:Gene Library; D006801:Humans; D015151:Immunoblotting; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000235:genetics* / Q000276:immunology*; D007074:Immunoglobulin G / Q000032:analysis / Q000235:genetics / Q000276:immunology; D008297:Male; D008459:Measles virus / Q000276:immunology*; D011233:Precipitin Tests; D013344:Subacute Sclerosing Panencephalitis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D018076:DNA, Complementary; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G