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

Stickel JS, Stickel N, Hennenlotter J, Klingel K, Stenzl A, Rammensee HG, Stevanović S - BMC Urol (2011)

Bottom Line: 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.Our results raise hopes for improving the success and effectiveness of future immunotherapeutic concepts for metastatic RCC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Immunology, Institute for Cell Biology, University of Tübingen, Germany.

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.

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Related in: MedlinePlus

Comparison TILs vs. no TILs. Amounts of immunopurified HLA class I compared between primary tumors containing TILs and tumor tissue without TILs. Yields were normalized to 1 g of tissue. The mean value of the normalized HLA yield of the respective tissue type is given plus standard error of the mean (SEM). Statistical significance of the differences between tissues is reflected in p values determined by the Student`s t-test and indicated in the histogram.
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Figure 2: Comparison TILs vs. no TILs. Amounts of immunopurified HLA class I compared between primary tumors containing TILs and tumor tissue without TILs. Yields were normalized to 1 g of tissue. The mean value of the normalized HLA yield of the respective tissue type is given plus standard error of the mean (SEM). Statistical significance of the differences between tissues is reflected in p values determined by the Student`s t-test and indicated in the histogram.

Mentions: In order to rule out the possibility that tumor infiltrating lymphocytes (TILs) have a strong impact on quantitative HLA analysis by Edman degradation, we compared primary RCCs from which TILs could be cultured with tumors from which no TILs could be isolated and cultured (Figure 2). The mean amount of HLA class I was 230.0 ± 55.9 pmol/g recovered from RCC with TILs (n = 11) compared to 176.0 ± 34.6 pmol/g from tumors without TILs (n = 6). This comparison shows that the HLA amount in tumors containing TILs is slightly higher than in tumors without TILs. However, statistical analysis showed no significance; the p value of 0.51 suggests a nearly identical HLA yield on both tumor groups.


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

Stickel JS, Stickel N, Hennenlotter J, Klingel K, Stenzl A, Rammensee HG, Stevanović S - BMC Urol (2011)

Comparison TILs vs. no TILs. Amounts of immunopurified HLA class I compared between primary tumors containing TILs and tumor tissue without TILs. Yields were normalized to 1 g of tissue. The mean value of the normalized HLA yield of the respective tissue type is given plus standard error of the mean (SEM). Statistical significance of the differences between tissues is reflected in p values determined by the Student`s t-test and indicated in the histogram.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3037347&req=5

Figure 2: Comparison TILs vs. no TILs. Amounts of immunopurified HLA class I compared between primary tumors containing TILs and tumor tissue without TILs. Yields were normalized to 1 g of tissue. The mean value of the normalized HLA yield of the respective tissue type is given plus standard error of the mean (SEM). Statistical significance of the differences between tissues is reflected in p values determined by the Student`s t-test and indicated in the histogram.
Mentions: In order to rule out the possibility that tumor infiltrating lymphocytes (TILs) have a strong impact on quantitative HLA analysis by Edman degradation, we compared primary RCCs from which TILs could be cultured with tumors from which no TILs could be isolated and cultured (Figure 2). The mean amount of HLA class I was 230.0 ± 55.9 pmol/g recovered from RCC with TILs (n = 11) compared to 176.0 ± 34.6 pmol/g from tumors without TILs (n = 6). This comparison shows that the HLA amount in tumors containing TILs is slightly higher than in tumors without TILs. However, statistical analysis showed no significance; the p value of 0.51 suggests a nearly identical HLA yield on both tumor groups.

Bottom Line: 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.Our results raise hopes for improving the success and effectiveness of future immunotherapeutic concepts for metastatic RCC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Immunology, Institute for Cell Biology, University of Tübingen, Germany.

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.

Show MeSH
Related in: MedlinePlus