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Cloning the Antibody Response in Humans with Inflammatory Central Nervous System Disease: Analysis of the Expressed IgG Repertoire in Subacute Sclerosing Panencephalitis Brain Reveals Disease-Relevant Antibodies That Recognize Specific Measles Virus Antigens¹

Mark P. Burgoon^*, Gregory P. Owens^*, Tracey Smith-Jensen^*, David Walker^* and Donald H. Gilden^2,*,

Departments of ^* Neurology and Microbiology, University of Colorado Health Sciences Center, Denver, CO 80262.

The presence of increased IgG in the brains of humans with infectious and inflammatory CNS diseases of unknown etiology such as multiple sclerosis may be a clue to the cause of disease. For example, the intrathecally synthesized oligoclonal bands (OGBs) in diseases such as subacute sclerosing panencephalitis (SSPE) or cryptococcal meningitis have been shown to represent Ab directed against the causative agents, measles virus (MV) or Cryptococcus neoformans, respectively. Using SSPE as a model system, we have developed a PCR-based strategy to analyze the repertoire of IgG V region sequences expressed in SSPE brain. We observed abnormal expression of germline V segments, overrepresentation of particular sequences that correspond to the oligoclonal bands, and substantial somatic mutation of most clones from the germline, which, taken together, constitute features of Ag-driven selection in the IgG response. Using the most abundant or most highly mutated H chain and or lambda L chain sequences in various combinations, we constructed functional Abs in IgG mammalian expression vectors. Three Abs specifically stained MV-infected cells. One Ab also stained cells transfected with the MV nucleoprotein, and a second Ab stained cells transfected with the MV-fusion protein. This technique demonstrates that functional Abs produced from putative disease-relevant IgG sequences can be used to recognize their corresponding Ags.

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