Targeting of tumor associated antigens in renal cell carcinoma using proteome-based analysis and their clinical significance

Abstract

The suitability of proteome-based strategies for the targeting of tumor-associated markers along with further analysis regarding their clinical significance were investigated in human renal cell carcinoma (RCC). The immunogenic protein expression profile of normal kidney and RCC cell lines was studied by proteome analysis combined with immunoblotting using sera from healthy donors and RCC patients, also termed PROTEOMEX. Employing this approach, a series of proteins reactive with either RCC patient sera and/or reactive with control sera were identified by microanalysis of tryptic peptides. Some of these candidate antigens represent members of the cytoskeletal family, such as cytokeratins, in particular cytokeratin 8, cytoskeletal tropomyosin, F-actin capping protein, gamma-actin, stathmin, tubulin-alpha, tubulin-beta and vimentin. The expression pattern and clinical significance of three of these antigens, namely cytokeratin 8, stathmin and vimentin, were further analyzed in a large series of surgically removed RCC lesions of distinct subtypes. A heterogeneous expression pattern of cytokeratin 8, stathmin and vimentin was demonstrated in the different RCC subtypes. All epithelial cells of the autologous normal kidney showed a strong cytokeratin 8 staining pattern, whereas they totally lack vimentin expression. Stathmin was expressed in 10% of tubule cells. In conclusion, PROTEOMEX could be employed for the identification of tumor-associated antigens of the cytoskeleton which are differentially expressed in RCC of distinct subtypes as well as in normal renal epithelium.

Application of immunoproteomics to leptospirosis: towards clinical diagnostics and vaccine discovery

“…Some of the identified proteins, i.e. DnaK (spot 1), fructose-bisphosphate aldolase (spots 10 and 11), dihydrolipoamide acyltransferase (spot 19), and conserved hypothetical protein (spots 23 and 24) were immunoreactive only with IgG, whereas the others Figure 1. IgM-immunoreactive proteins.…”

“…renal cell carcinoma. These include the 'Serological and proteomic evaluation of antibody responses' (SPEAR) in the study by Unwin et al [17], 'Serological proteomic analysis' (SERPA) in the study by Klade et al [18] and 'Proteomex' in reports by Seliger and colleagues [19][20][21]. These techniques were named differently but all have similar analytical strategy that combines 2-D PAGE, MS identification, and immunoblotting, using cancer tissues as the antigens and sera from patients as the antibodies.…”

A novel MHC-associated Proteinase 3 peptide isolated from primary chronic myeloid leukaemia cells further supports the significance of this antigen for the immunotherapy of myeloid leukaemias

“…14 Moreover, the detection of a humoral response against Stathmin in renal cell carcinoma (RCC) patients would indicate that this protein is immunogenic in other tumors. 15 In addition to the peptide from Stathmin, an MHC ligand was also identified from an additional protein to which a humoral response has been previously detected in tumour patients, Elongation factor 1-alpha 1. This has been identified in a cutaneous T-cell lymphoma (CTCL).…”

“…CML-85 was A*0201, A*1101, B*1501 and B*27. Karyotyping of CML-83 showed: 46, XX, ins(4;7)(q21;q22q32), t(7;9;22)(q32;q34;q11) [10], and for CML-85: 46, XY, t(9;22) (q34;q11) [15].…”

Immunoproteomics technologies in the discovery of autoantigens in autoimmune diseases

“…The most common method used for profiling autoreactivity of patient sera and identification of antigens in autoimmune diseases is referred to as serological proteome analysis (SERPA) (4) or PROTEOMEX (5). In this method, whole tissue/cell protein preparations containing potential autoantigens are run on 2-dimensional electrophoresis (2DE) gel in triplicate ( Figure 1A).…”