PMID- 8574469
OWN - NLM
STAT- MEDLINE
DCOM- 19960314
LR  - 20191101
IS  - 1067-5582 (Print)
IS  - 1067-5582 (Linking)
VI  - 18
IP  - 2
DP  - 1995 Aug
TI  - The use of Epstein-Barr virus-transformed B lymphocyte cell lines in a 
      peptide-reconstitution assay: identification of CEA-related HLA-A*0301-restricted 
      potential cytotoxic T-lymphocyte epitopes.
PG  - 77-85
AB  - In the development of cytotoxic T lymphocyte (CTL)-mediated immunotherapy, the 
      identification of CTL epitopes is of crucial importance. Binding of a peptide to 
      major histocompatibility complex (MHC) class I molecules is one of the 
      prerequisites for its function as a CTL epitope. We describe the technique, 
      validation, and application of a simple cellular assay, intended for the 
      screening of peptides for binding, that can be applied to any human leukocyte 
      antigen (HLA) allele. Reconstitution of peptides in MHC class I molecules after 
      elution by acid treatment was previously shown to be possible in specially 
      engineered cell lines expressing only one type of MHC class I, and was applied 
      for the HLA-A*0201 allele. We now report the optimal conditions for application 
      of this type of binding assay to the HLA-A*0301 allele. The adaptations that were 
      necessary to make the technique operational for HLA-A*0301 are shown in detail. 
      These consisted of lowering the pH during acid treatment to 2.9 and lengthening 
      the duration of elution to 90 s. Furthermore, immediate aspiration of eluted 
      peptides appeared to be essential for this allele. We found also that the use of 
      Epstein-Barr virus (EBV)-transformed B cell lines (B-LCL) yields results similar 
      to those of the use of cell lines expressing only one specific MHC class I 
      allele. Homozygosity for the desired HLA allele improves the sensitivity of the 
      assay, but heterozygous cells can also be employed. Finally, we applied this 
      technique to a search for HLA-A*0301 binding peptides derived from 
      carcinoembryonic antigen (CEA). Of a set of 34 CEA-specific peptides that fit 
      with a specified HLA-A*0301-binding motif, we identified a set of six peptides 
      with high binding affinity to this allele. These peptides can be regarded as 
      potential CTL epitopes.
FAU - Bremers, A J
AU  - Bremers AJ
AD  - Department of Surgery, University Hospital Leiden, The Netherlands.
FAU - van der Burg, S H
AU  - van der Burg SH
FAU - Kuppen, P J
AU  - Kuppen PJ
FAU - Kast, W M
AU  - Kast WM
FAU - van de Velde, C J
AU  - van de Velde CJ
FAU - Melief, C J
AU  - Melief CJ
LA  - eng
GR  - R01 CA579333/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Immunother Emphasis Tumor Immunol
JT  - Journal of immunotherapy with emphasis on tumor immunology : official journal of 
      the Society for Biological Therapy
JID - 9418950
RN  - 0 (Carcinoembryonic Antigen)
RN  - 0 (Epitopes)
RN  - 0 (HLA-A Antigens)
SB  - IM
MH  - Amino Acid Sequence
MH  - B-Lymphocytes/*virology
MH  - Biological Assay
MH  - Carcinoembryonic Antigen/*immunology
MH  - Cell Line, Transformed
MH  - Epitopes/*blood
MH  - HLA-A Antigens/*blood/genetics
MH  - Herpesvirus 4, Human/*physiology
MH  - Heterozygote
MH  - Homozygote
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Immunotherapy
MH  - Molecular Sequence Data
MH  - *Peptide Biosynthesis
MH  - T-Lymphocytes, Cytotoxic/*immunology
EDAT- 1995/08/01 00:00
MHDA- 1995/08/01 00:01
CRDT- 1995/08/01 00:00
PHST- 1995/08/01 00:00 [pubmed]
PHST- 1995/08/01 00:01 [medline]
PHST- 1995/08/01 00:00 [entrez]
AID - 10.1097/00002371-199508000-00001 [doi]
PST - ppublish
SO  - J Immunother Emphasis Tumor Immunol. 1995 Aug;18(2):77-85. doi: 
      10.1097/00002371-199508000-00001.