Increased IgG and oligoclonal bands (OGBs) in the CSF and brains of patients with chronic inflammatory or demyelinating CNS disease provide clues to the nature of disease. OGBs appear almost exclusively in chronic infectious diseases of the CNS, and are antibody directed against the disease-causing organism, thus providing a rationale for our hypothesis that in chronic CNS inflammatory or demyelinating diseases of unknown cause, the OGBs target the antigens that trigger disease. We have analyzed the sequences of intrathecally synthesized IgG in subacute sclerosing panencephalitis (SSPE), a fatal chronic measles virus infection of the brain, and demonstrated features of antigen-driven selection. We produced recombinant IgG from these sequences, selected disease-relevant recombinant IgG from phage-displayed Fab libraries, and demonstrated their reactivities to the causative measles virus. We will use SSPE as an experimental paradigm to develop strategies and techniques to identify disease-relevant IgG and their corresponding antigens in inflammatory or demyelinating CNS disease of unknown cause. We will synthesize recombinant IgG from candidate SSPE brain-derived sequences and identify disease-relevant antibody by direct analysis in immunostaining or ELISA with SSPE brain, or display them on phage surfaces to select the appropriate IgG by biopanning on SSPE brain. We will also identify the IgG sequences expressed by individual resident B cells in SSPE brain by single-cell PCR. Finally, we will develop strategies to detect very rare antigens in situ by immunoblotting, immunoprecipitation, or by selection from phage-displayed antigen and peptide libraries. We are uniquely prepared to carry out these studies because we have pathologically-verified SSPE brains, and have already demonstrated an expertise to use disease-relevant recombinant IgG to identify their targets. This paradigm will provide a careful and methodological approach to elucidate the humoral immune response in the human CNS. It will also allow the creation of exquisitely sensitive immunologic techniques to identify disease-relevant IgG and their disease-triggering antigens in other inflammatory or demyelinating CNS diseases of unknown cause, including multiple sclerosis, acute disseminated encephalomyelitis, sarcoidosis, and the CNS vasculitides. Determining the cause of these devastating neurologic diseases will have profound implications not only for early diagnosis but also for prevention of disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS041549-03
Application #
6684095
Study Section
Special Emphasis Panel (ZRG1-BDCN-4 (01))
Program Officer
Utz, Ursula
Project Start
2001-12-15
Project End
2005-11-30
Budget Start
2003-12-01
Budget End
2004-11-30
Support Year
3
Fiscal Year
2004
Total Cost
$218,619
Indirect Cost
Name
University of Colorado Denver
Department
Neurology
Type
Schools of Medicine
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
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Yu, Xiaoli; Barmina, Olga; Burgoon, Mark et al. (2009) Identification of measles virus epitopes using an ultra-fast method of panning phage-displayed random peptide libraries. J Virol Methods 156:169-73
Bennett, Jeffrey L; Haubold, Kurt; Ritchie, Alanna M et al. (2008) CSF IgG heavy-chain bias in patients at the time of a clinically isolated syndrome. J Neuroimmunol 199:126-32
Yu, Xiaoli; Burgoon, Mark P; Shearer, Andrew J et al. (2007) Characterization of phage peptide interaction with antibody using phage mediated immuno-PCR. J Immunol Methods 326:33-40
Owens, Gregory P; Winges, Kimberly M; Ritchie, Alanna M et al. (2007) VH4 gene segments dominate the intrathecal humoral immune response in multiple sclerosis. J Immunol 179:6343-51

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