Identification of β-Tubulin Isoforms as Tumor Antigens in Neuroblastoma
2000
There is currently
substantial interest in the identification of human tumor antigens for
diagnosis and immunotherapy of cancer. We have implemented a proteomic
approach for the identification of tumor proteins that elicit a humoral
response in cancer patients, which we have applied to neuroblastoma.
Proteins from neuroblastoma tumors and cell lines were separated by
two-dimensional PAGE and transferred to poly(vinylidene
difluoride) membranes. Sera from 23 newly diagnosed patients
with neuroblastoma, from 12 newly diagnosed children with other solid
tumors, and from 13 normal individuals were screened for IgG and IgM
autoantibodies against neuroblastoma proteins by means of Western blot
analysis. Sera from 11 patients with neuroblastoma and from 1 patient
with a primitive neuroectodermal tumor, but none of the other controls
exhibited IgG-based reactivity against a protein constellation with an
estimated M r 50,000.
NH 2 -terminal sequence and mass spectrometric analysis
identified the major constituents of this constellation as β-tubulin
isoforms I and III. The IgG antibodies were additionally characterized
to be of the subclass IgG1. Neuroblastoma patient sera that contained
anti-β-tubulin IgG antibodies also contained IgM antibodies specific
against the full-length β-tubulin molecule and against COOH-terminalβ
-tubulin cleavage products. Neuroblastoma patient sera that reacted
with β-tubulin I and III isoforms in neuroblastoma tissues did not
react with βtubulin I and III isoforms found in normal brain
tissue. Our findings indicate the occurrence of β-tubulin peptides in
neuroblastoma, which are immunogenic. The occurrence of immunogenic
peptides in neuroblastoma may have utility in diagnosis and in
immunotherapy of this aggressive childhood tumor.
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