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Mapping and expression pattern analysis of key components of the major histocompatibility complex class I antigen processing and presentation pathway in a representative human renal cell carcinoma cell line
Author(s) -
Lichtenfels Rudolf,
Ackermann Angelika,
Kellner Roland,
Seliger Barbara
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200105)22:9<1801::aid-elps1801>3.0.co;2-i
Subject(s) - antigen processing , immunotherapy , major histocompatibility complex , renal cell carcinoma , biology , antigen , antigen presentation , mhc class i , microbiology and biotechnology , cell culture , cancer research , t cell , immunology , medicine , immune system , pathology , genetics
Renal cell carcinoma (RCC) represent approximately 5% of all cancer deaths. At the time of presentation, over 50% of the patients have already developed locally advanced or metastatic disease with five‐year survival rates of less than 20%. Although relative resistant to conventional regimens, RCC are partially susceptible to T cell‐based immunotherapy. To further develop this treatment modality, two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) was applied for both the mapping of the key components of the major histocompatibility complex (MHC) class I antigen processing and presentation machinery (APM) and the characterization of the constitutive and cytokine‐regulated protein expression profiles in a representative human RCC cell line. The latter aspect is based on the fact, that the expression level of some of the APM components can be altered in response to interferon (IFN)‐γ treatment. Total cell lysates from untreated and IFN‐γ‐treated tumor cells were separated on 2‐D PAGE gels using broad range immobilized pH gradient (IPG) strips. Serial Western blot analyses using sets of APM‐specific antibodies were performed to target the relevant protein spots. Protein verification was mostly accomplished via peptide mass fingerprinting using matrix‐assisted laser desorption/ionization‐mass spectrometry (MALDI‐MS). To date, the majority of the APM‐related components have been identified and mapped. In addition, the different protein expression profiles of untreated and IFN‐γ‐treated RCC cells are under investigation.