Abstract
BackgroundSera from children with post-varicella infections have autoantibodies that react with centrosomes in brain and tissue culture cells. We investigated the sera of children with infections and post-varicella ataxia and related conditions for reactivity to five recombinant centrosome proteins: γγ-enolase, pericentrin, ninein, PCM-1, and Mob1.MethodsSera from 12 patients with acute post-varicella ataxia, 1 with post-Epstein Barr virus (EBV) ataxia, 5 with uncomplicated varicella infections, and other conditions were tested for reactivity to cryopreserved cerebellum tissue and recombinant centrosome proteins. The distribution of pericentrin in the cerebellum was studied by indirect immunofluorescence (IIF) using rabbit antibodies to the recombinant protein. Antibodies to phospholipids (APL) were detected by ELISA.ResultsEleven of 12 children with post-varicella ataxia, 4/5 children with uncomplicated varicella infections, 1/1 with post-EBV ataxia, 2/2 with ADEM, 1/2 with neuroblastoma and ataxia, and 2/2 with cerebellitis had antibodies directed against 1 or more recombinant centrosome antigens. Antibodies to pericentrin were seen in 5/12 children with post-varicella ataxia but not in any of the other sera tested. IIF demonstrated that pericentrin is located in axons and centrosomes of cerebellar cells. APL were detected in 75% of the sera from children with post-varicella ataxia and 50% of children with varicella without ataxia and in none of the controls.ConclusionThis is the first study to show the antigen specificity of anti-centrosome antibodies in children with varicella. Our data suggest that children with post-varicella ataxia have unique autoantibody reactivity to pericentrin.
Highlights
Sera from children with post-varicella infections have autoantibodies that react with centrosomes in brain and tissue culture cells
We investigated the possibility that autoantibodies to specific centrosome proteins are associated with a subset of children with post-varicella acute cerebellar ataxia
Eleven of 12 sera from children with post-varicella ataxia had IgG antibodies that reacted with the centrosome of formalin-fixed HeLa cells in an indirect immunofluorescence (IIF) assay (Figure 1)
Summary
Patients & Sera Children with chicken pox and subsequent cerebellar ataxia were included in this study [8,9,9]. Indirect immunofluorescence (IIF) Conventional IF was performed on commercially available HEp-2 cells (Immuno Concepts Inc., Sacramento, CA) and formalin-fixed HeLa cells using a fluorescein-conjugated goat anti-human IgG (light and heavy chain) as previously described [20]. AFiugtuoraent1ibodies to centrosomes Autoantibodies to centrosomes Indirect immunofluorescence illustrating antibodies to centrosomes as seen in sera of children with post-varicella ataxia on formalin fixed HeLa cell substrate. Bacterial lysates that expressed the recombinant proteins were resuspended in sodium dodecyl sulfate sample buffer and denatured by boiling for 5 minutes as previously described [23]. An anti-human immunoglobulin (IgG,A,M) secondary antibody conjugated to horseradish peroxidase was added and incubated for 15 minutes at room temperature. The reactivities were read at an absorbance of 450 nm on a Biomek 1000 (Beckman) and cutoffs based on graded control positive and negative control sera provided with the kit
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