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Structure and function of ETAA16: a novel cell surface antigen in Ewing’s tumours

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Abstract

Immunoscreening of an Ewing’s family of tumour (EFT)-derived cDNA library using formerly described EFT-specific antibodies led to the isolation of a 3.5 kb cDNA, named Ewing’s tumour-associated antigen 16 (ETAA16). The ETAA16 cDNA shows no homology to any functionally characterised human gene. Only a bovine cDNA expressed in bovine testis and hepatocytes is functionally characterised as it encodes for a junction plaque associated protein and showed a homology of 69.9% at amino acid level to ETAA16. The human cDNA encodes for a 926 amino acid tumour antigen with a calculated molecular weight of 103 kDa. The epitope of the ETAA16-specific antibody, Ak16, covers the central region of the protein which is part of an extra cellular domain. The human ETAA16 gene locus has been assigned to chromosome 2p13-15 by FISH analyses and is confirmed by the human genome sequencing project. As demonstrated by flow cytometry, the cell surface expression of ETAA16 antigen is restricted to ET cell lines and not expressed on other small blue round cell tumours or other kind of tumour. RT-PCR analysis revealed a high expression of ETAA16 in brain, liver and kidney while lung and heart were negative. Immunohistochemistry showed an intracellular expression of ETAA16 in different kind of non-Ewing tumour tissues. These results suggest that ETAA16 may function as a tumour-specific cell surface antigen in EFTs.

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Abbreviations

DAB:

Diaminobenzidine

EFT:

Ewing’s family of tumours

ET:

Ewing’s tumour

ETAA16:

Ewing’s tumour-associated antigen 16

ETS:

E26 transformation specific

EST:

Expressed sequence tag

EWS:

Ewing’s sarcoma

FISH:

Fluorescence in situ hybridisation

FLI-1:

Friend leukemia integration site-1

MAB:

Monoclonal antibody

MFI:

Mean fluorescence intensity

ORF:

Open reading frame

PALS:

Periarteriolar lymphoid sheath

PNET:

Primitive neuroectodermal tumour

UTR:

Untranslated region

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft DI 747/3–1; Di747/3–2 and the Krebshilfe, Dr. Mildred Scheel Stiftung: 10–1859/Di, W26/94/Schn2 and the Elterninitiative Kinderkrebsklinik e.V. The authors thank Professor Horst Hameister, Department of Human Genetics, University of Ulm for performing the FISH analyses and his contribution in data analysis; stud. Biol. Stefanie Borkens Heinrich Heine University Düsseldorf for excellent technical assistance, Prof. Guido Reifenberger and Prof. Christopher Poremba, Institut for Pathology University Medical Center Düsseldorf for providing various cryopreserved tumour tissues. The authors also wish to thank Bettina Stahl (Anaesthesiology, Ulm) and Whilhelma Langmann (Department of Anatomy, Bonn) for their help with the immunohistochemical staining and also Dr. Kay Hofmann, Memorec, Cologne, Germany, for the bioinformatics analysis.

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Author notes

  1. A. Borowski

    Present address: Department of Anatomy, University of Bonn, Nussallee 10, 53115, Bonn, Germany

Authors and Affiliations

  1. Department of Paediatric Oncology, Haematology and Immunology, University of Dusseldorf, Moorenstrasse 5, 40225, Düsseldorf, Germany

    A. Borowski, U. Dirksen, L. Lixin, R. L. Shi & U. Göbel

  2. Section of Experimental Anaesthesiology, Department of Clinical Anaesthesiology, University of Ulm, Steinhoevelstrasse 9, 89075, Ulm, Germany

    E. M. Schneider

  3. Child and Youth, Department of Paediatric Haematlogy and Oncology, University of Muenster, Albert Schweizerstr. 33, 48129, Muenster, Germany

    U. Dirksen

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  1. A. Borowski
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Correspondence to A. Borowski.

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Borowski, A., Dirksen, U., Lixin, L. et al. Structure and function of ETAA16: a novel cell surface antigen in Ewing’s tumours. Cancer Immunol Immunother 55, 363–374 (2006). https://doi.org/10.1007/s00262-005-0017-6

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  • DOI: https://doi.org/10.1007/s00262-005-0017-6

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