Affiliation:
1. Departments
of Neurology
2. Ophthalmology
3. Microbiology,University of Colorado Health Sciences Center,
Denver, Colorado 80262
Abstract
ABSTRACT
Infectious
and inflammatory diseases of the CNS are often characterized by a
robust B-cell response that manifests as increased intrathecal
immunoglobulin G (IgG) synthesis and the presence of oligoclonal bands.
We previously used laser capture microdissection and single-cell PCR to
analyze the IgG variable regions of plasma cells from the brain of a
patient with subacute sclerosing panencephalitis (SSPE). Five of eight
human IgG1 recombinant antibodies (rAbs) derived from SSPE brain plasma
cell clones recognized the measles virus (MV) nucleocapsid protein,
confirming that the antibody response in SSPE targets primarily the
agent causing disease. In this study, as part of our work on antigen
identification, we used four rAbs to probe a random phage-displayed
peptide library to determine if epitopes within the MV nucleocapsid
protein could be identified with SSPE brain rAbs. All four of the SSPE
rAbs enriched phage-displayed peptide sequences that reacted
specifically to their panning rAb by enzyme-linked immunosorbent assay.
BLASTP searches of the NCBI protein database revealed clear homologies
in three peptides and different amino acid stretches within the 65
C-terminal amino acids of the MV nucleocapsid protein. The
specificities of SSPE rAbs to these regions of the MV nucleocapsid
protein were confirmed by binding to synthetic peptides or to short
cDNA expression products. These results indicate the feasibility of
using peptide screening for antigen discovery in central nervous system
inflammatory diseases of unknown etiology, such as multiple sclerosis,
neurosarcoidosis, or Behcet's
syndrome.
Publisher
American Society for Microbiology
Subject
Virology,Insect Science,Immunology,Microbiology
Cited by
17 articles.
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