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Gatz, S.A. ; Pohla, H.* ; Schendel, D.J.

A PCR-SSP method to specifically select HLA-A*0201 individuals for immunotherapeutic studies.

Tissue Antigens 55, 1-16 (2000)
Open Access Green as soon as Postprint is submitted to ZB.
LA-A*0201 is an important restriction element for peptide presentation to T cells in disease and cancer. Mutation studies and analyses using cytotoxic T lymphocytes have shown the functional relevance of sub type-specific differences in HLA-A2 molecules for peptide binding and T-cell receptor recognition. Therefore, many immunotherapeutic studies need to accurately select HLA-A*0201-positive individuals. We designed an easy, robust approach based on the polymerase chain reaction using sequence-specific primers (PCR-SSP) to specifically distinguish A*0201-positive individuals from other HLA-A2 subtypes described to date. The first step includes reactions that give information whether the sample donor is HLA-A2 and, if so, whether the individual is homozygous or heterozygous for HLA-A2. Further, it is determined whether the sample has an HLA-A*020S or an HLA=A*0201 sequence at the corresponding position in exon 4. Samples that may contain an HLA-A*0201 allele according to the results of this first step are subtyped in a second step nested PCR. Here the strategy is focussed on the discrimination of HLA-A*0201 from the other subtypes by considering divergent nucleotide positions in two ways. One SSP combination amplifies the HLA-A*0201 sequence while a corresponding SSP combination specifically amplifies the subtype or group of subtypes differing from HLA-A*0201 at this position. Thus, at relevant polymorphic nucleotide positions the HLA-A*0201 sequence is both directly and indirectly confirmed. This strategy strongly enhances the reliability of the subtyping and allows better verification of HLA-A*0201-positive patient selection for clinical studies.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords DNA-typing; HLA-A*0201; HLA-A*02011; HLA-A*02 subtyping; immunotherapy; PCR-SSP
ISSN (print) / ISBN 0001-2815
e-ISSN 1399-0039
Journal Tissue Antigens
Quellenangaben Volume: 55, Issue: , Pages: 1-16 Article Number: , Supplement: ,
Publisher Wiley
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Molecular Immunology (IMI)