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Open Access Research article

Quantification of HLA class I molecules on renal cell carcinoma using Edman degradation

Juliane S Stickel1, Natalie Stickel3, Jörg Hennenlotter2, Karin Klingel4, Arnulf Stenzl2, Hans-Georg Rammensee1 and Stefan Stevanović1*

Author Affiliations

1 Department of Immunology, Institute for Cell Biology, University of Tübingen, Germany

2 Clinic for Urology, University of Tübingen, Germany

3 Department of Hematology Oncology, University of Freiburg, Germany

4 Department of Molecular Pathology, University of Tübingen, Germany

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BMC Urology 2011, 11:1  doi:10.1186/1471-2490-11-1

Published: 20 January 2011

Abstract

Background

Unimpaired HLA class I antigen presentation is a prerequisite for the recognition of tumor cells by cytotoxic T lymphocytes and thus essential for the success of anticancer immunotherapeutic concepts. Several approaches have been taken in the immunotherapy of metastatic renal cell carcinoma (RCC), however of limited success. HLA loss or down-regulation have often been reported and might interfere with immunotherapeutic approaches aimed at the recognition of HLA-presented peptides.

Methods

We employed a quantitative method of molecular analysis for the comparison of HLA amounts on primary tumor, normal kidney and metastases of RCC, using Edman degradation. We analyzed a series of 47 RCC samples including corresponding renal parenchyma, local lymph node metastases and distant metastases.

Results

Results of quantitative Edman degradation revealed significantly higher HLA yields on primary tumor and metastases compared to normal kidney tissue. This effect was shown not to result from infiltrating immune cells, since tumor-infiltrating lymphocytes had no influence on the overall HLA recovery from tumor tissue. Unexpectedly, we found a higher amount of HLA class I molecules on distant metastases compared to local lymph node metastases.

Conclusion

Edman degradation allows the direct quantitative comparison of HLA class I protein expression by tumor or normal tissue and metastases of RCC patients. Our results raise hopes for improving the success and effectiveness of future immunotherapeutic concepts for metastatic RCC.