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

J Neuroimmunol. 1999 Feb 1;94(1-2):204-11. doi: 10.1016/s0165-5728(98)00243-4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adolescent
  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology
  • Antibodies, Viral / immunology
  • Antibodies, Viral / isolation & purification*
  • Antibody Specificity
  • Antigens, Viral / analysis
  • Antigens, Viral / isolation & purification
  • Bacteriophages
  • Brain / immunology
  • Brain / virology
  • Cloning, Molecular
  • DNA, Complementary
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes
  • Gene Library
  • Humans
  • Immunoblotting
  • Immunoglobulin Fab Fragments / analysis
  • Immunoglobulin Fab Fragments / genetics*
  • Immunoglobulin Fab Fragments / immunology*
  • Immunoglobulin G / analysis
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology
  • Male
  • Measles virus / immunology*
  • Precipitin Tests
  • Subacute Sclerosing Panencephalitis / immunology*

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • DNA, Complementary
  • Epitopes
  • Immunoglobulin Fab Fragments
  • Immunoglobulin G