Laser-capture microdissection of plasma cells from subacute sclerosing panencephalitis brain reveals intrathecal disease-relevant antibodies.
Increased IgG and oligoclonal bands are found in cerebrospinal fluid of humans with chronic infectious CNS disease. Studies have shown that these oligoclonal bands are antibodies directed against the agent that causes disease. Laser-capture microdissection was used to isolate individual CD38+ plasma cells from the brain of a patient with subacute sclerosing panencephalitis, and single-cell RT-PCR was used to analyze individual IgG heavy and light chains expressed by each cell. Based on overrepresented IgG sequences, we constructed functional recombinant antibodies (recombinant IgGs) and determined their specificities. Five of eight recombinant IgGs recognized measles virus, the cause of subacute sclerosing panencephalitis. These results demonstrate that overrepresented IgG sequences in postmortem brains can be used to produce functional recombinant antibodies that recognize their target antigens. This strategy can be used to identify disease-relevant antigens in CNS inflammatory diseases of unknown etiology.
Burgoon
Mark P
MP
Department of Neurology, University of Colorado Health Sciences Center, 4200 East 9th Avenue, Denver, CO 80262, USA. mark.burgoon@uchsc.edu
Keays
Kathryne M
KM
Owens
Gregory P
GP
Ritchie
Alanna M
AM
Rai
Pradeep R
PR
Cool
Carlyne D
CD
Gilden
Donald H
DH
D036541:ADP-ribosyl Cyclase / Q000276:immunology; D051997:ADP-ribosyl Cyclase 1; D000293:Adolescent; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D001921:Brain / Q000276:immunology / Q000473:pathology*; D002460:Cell Line; D003587:Cytomegalovirus; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology*; D007834:Lasers; D008297:Male; D008459:Measles virus / Q000276:immunology; D008562:Membrane Glycoproteins; D042282:Microdissection; D010950:Plasma Cells / Q000276:immunology* / Q000473:pathology; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013344:Subacute Sclerosing Panencephalitis / Q000276:immunology* / Q000473:pathology* / Q000821:virology
D000888:Antibodies, Anti-Idiotypic; D015703:Antigens, CD; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1