Comparative study of peptides isolated from class I antigen groove of urological specimens. a further step toward the future of peptide therapy in bladder cancer patients.
BACKGROUND
The major histocompatibility (MHC) class I antigens act as associative molecules for interaction amongst immuno-competent cells. The grooves of class I antigens are normally loaded with self peptides of between 8 and 11 amino acids. However, when the cells transform to malignant state they may carry peptide(s) of non-self origin within these grooves. Using immuno-bead purification followed by high-performance liquid chromatography (HPLC), this study attempted to isolate peptides from class I antigens of various biological specimens


RESULTS
The combination of immuno-bead purification (BP) and HPLC was reliable for peptide isolation. Class I antigens and associated peptides could be isolated from normal peripheral blood leukocyte (PBL). Under the same conditions, the PBL yielded almost twice as much peptide as that of tumour cell lines. The HPLC profile of peptides (range of 8-10 amino acid residues) isolated from a bladder and a cervical tumour cell line showed unique features. In the case of the bladder line there were at least 22 peptides. In addition, the class I-associated peptides could also be isolated from kidney tumour fragments of three individuals. In each the isolated peptides showed a unique HPLC peak profile with some similarities as well as differences.


CONCLUSIONS
These data indicated a variation in the nature of peptides isolated from different specimens. The approach showed the feasibility of preparing peptide(s) from a relatively small number of cells. The data also showed that peptide isolation could also be carried out from tumour tissue biopsies paving the way for the future of peptide vaccination in cancer patients following the identification of putative tumour-specific replicate(s).
Torabi-Pour
N
N
Uro/Oncology, Research Unit, Bart's and The London Trust, London, UK. n.torabi-pour@mds.qmw.ac.uk
Nouri
A M E
AM
Saffie
R
R
Oliver
R T D
RT
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D018014:Gene Transfer Techniques; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D010455:Peptides / Q000302:isolation & purification*; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000737:chemistry / Q000276:immunology* / Q000473:pathology
D015395:Histocompatibility Antigens Class I; D010455:Peptides