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Identification of HLA a*0201 glioblastoma
multiforme cell lines for immunotherapy by PCR-SSP and DNA sequencing
Wu AH, Hall WA, Low WC.
Department of Neurosurgery, University of Minnesota Medical
School, Minneapolis, MN 55455, USA.
Most tumor specific antigens characterized to date are restricted by HLA a*0201,
which is the major HLA subtype in many ethnic groups.
Cancer cells that express tumor antigens in association with the HLA a*0201
subtype have been shown to be responsive to various immunotherapies.
We therefore sought to identify glioma cell lines that also express this HLA
subtype and determine whether they had the molecular properties needed for
tumor-peptide presentation.
The HLA a*0201 allele was identified with PCR using sequence-specific primers
followed by DNA sequencing.
With this method, we screened 15 glioma cell lines to determine if they were of
the HLA a*0201 genotype.
Glioma cell lines that express the HLA a*0201 subtype were further studied for
the expression of MHC class I and beta-2-microglobulin (beta2m) molecules by
flow cytometry, and peptide presentation molecules TAP-1, TAP-2, and tapasin by
RT-PCR.
We identified six out of fifteen cell lines that were of the HLA a*0201
subtype.
These cell lines are U87, T98, U373, U138, CRL2365 and UMN-4.
All these six cell lines exhibited high levels of MHC class I and beta2m
molecules.
In addition, these cell lines all expressed molecules required for peptide
presentation as shown by the presence of peptide presentation-related molecules
TAP-1, TAP-2 and tapasin.
The identification of glioma cell lines that express the HLA a*0201 subtype
along with the necessary molecules for peptide-presentation will enable their
use in developing new immunotherapeutic approaches for treating brain
tumors.
The method used to identify HLA a*0201 glioma cells is rapid and inexpensive,
and suitable for screening tumor cells.
PMID: 15015764 [PubMed - indexed for MEDLINE]
Source: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15015764
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