PMID- 12372891
OWN - NLM
STAT- MEDLINE
DCOM- 20030124
LR  - 20180213
IS  - 0042-1138 (Print)
IS  - 0042-1138 (Linking)
VI  - 69
IP  - 3
DP  - 2002
TI  - Isolation of human leukocyte antigen (HLA)-associated peptide(s) in the absence 
      of HLA-restricted specific cytolytic T lymphocytes.
PG  - 219-26
AB  - BACKGROUND/METHODS: In this study, immunobead purification, dot-blot, 
      immunocytochemical staining, and SDS-PAGE techniques in combination with 
      high-performance liquid chromatography were used to isolate human leukocyte 
      antigen (HLA) class I antigens and associated peptides from a bladder tumour cell 
      line (Fen) before and after gene transfection. RESULTS: The results showed that: 
      (1) Transfection of the class I negative Fen cell line with normal 
      beta-microglobulin (beta(2)-m) gene resulted in the restoration of missing class 
      I antigens. (2) The intact class I antigens could be isolated from lysate of the 
      beta(2)-m gene transfected cells using Sepharose CNBr-W6/32 beads. (3) 
      Dissociation of class I antigens from beads and analysis by the SDS-PAGE showed 
      the presence of both free heavy and light chains of class I antigens. (4) More 
      than 22 class I-associated peptides with a molecular weight of 700-3,000 daltons 
      could be isolated from W6/32-loaded beads but only from lysate of HLA-positive 
      Fen cell line. The data also showed that 1 x 10(6) of positive Fen cells 
      contained about 200 microg total protein of which about 0.10 microg was class I 
      and about 2 ng was class I-associated peptides. CONCLUSIONS: These findings 
      demonstrated that the gene transfection approach could be used to restore missing 
      class I antigens on an otherwise class I negative bladder tumour cell line. The 
      results also showed the feasibility of using above techniques for isolation of 
      HLA-associated peptides. These approaches may provide a realistic possibility for 
      identification of putative tumour-specific peptide(s) from tumour specimens with 
      the long-term aim to use such peptide(s) for immunotherapy in cancer patients.
CI  - Copyright 2002 S. Karger AG, Basel
FAU - Torabi-Pour, N
AU  - Torabi-Pour N
AD  - Uro/Oncology Research Unit, Department of Medical Oncology and Urology, The Royal 
      London Trust, UK. n.torabi-pour@mds.qmw.ac.uk
FAU - Nouri, A M E
AU  - Nouri AM
FAU - Saffie, R
AU  - Saffie R
FAU - Morrow, W J W
AU  - Morrow WJ
FAU - Oliver, R T D
AU  - Oliver RT
LA  - eng
PT  - Journal Article
PL  - Switzerland
TA  - Urol Int
JT  - Urologia internationalis
JID - 0417373
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (Peptides)
SB  - IM
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Histocompatibility Antigens Class I/*isolation & purification
MH  - Humans
MH  - Peptides/*isolation & purification
MH  - Protein Binding
MH  - Sensitivity and Specificity
MH  - T-Lymphocytes/chemistry/*immunology
MH  - Transfection
MH  - Tumor Cells, Cultured
MH  - Urinary Bladder Neoplasms/*immunology/pathology
EDAT- 2002/10/10 04:00
MHDA- 2003/01/25 04:00
CRDT- 2002/10/10 04:00
PHST- 2002/10/10 04:00 [pubmed]
PHST- 2003/01/25 04:00 [medline]
PHST- 2002/10/10 04:00 [entrez]
AID - uin69219 [pii]
AID - 10.1159/000063943 [doi]
PST - ppublish
SO  - Urol Int. 2002;69(3):219-26. doi: 10.1159/000063943.