Visualization of p53(264-272)/HLA-A*0201 complexes naturally presented on tumor cell surface by a multimeric soluble single-chain T cell receptor.
Intracellular Ags are processed into small peptides that are presented on cell surfaces in the context of HLA class I molecules. These peptides are recognized by TCRs displayed by CD8+ T lymphocytes (T cells). To date, direct identification and quantitation of these peptides has relied primarily on mass spectrometry analysis, which is expensive and requires large quantities of diseased tissues to obtain useful results. Here we demonstrate that multimerization of a soluble single-chain TCR (scTCR), recognizing a peptide from p53 presented in the context of HLA-A2.1, could be used to directly visualize and quantitate peptide/MHC complexes on unmanipulated human tumor cells. Tumor cells displaying as few as 500 peptide/MHC complexes were readily detectable by flow cytometry. The scTCR/multimers exhibited exquisite recognition capability and could distinguish peptides differing in as little as a single amino acid. We also demonstrate that scTCR/multimers could specifically stain human tumors generated in mice, as well as tumors obtained from patient biopsies. Thus, scTCR/multimers represent a novel class of immunostaining reagents that could be used to validate, quantitate, or monitor epitope presentation by cancer cells.
Altor BioScience Corp., Miramar, FL 33025, USA.
Xiaoyun
X
Zhu
Heather J
HJ
Belmont
Shari
S
Price-Schiavi
Bai
B
Liu
Hyung-il
HI
Lee
Marilyn
M
Fernandez
Richard L
RL
Wong
Janette
J
Builes
Peter R
PR
Rhode
Hing C
HC
Wong
D000368:Aged; D000818:Animals; D017951:Antigen Presentation; D016466:CHO Cells; D019731:Carbon-Nitrogen Ligases / Q000737:chemistry / Q000235:genetics; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D006224:Cricetinae; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics; D005260:Female; D015234:HLA-A Antigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D015789:HLA-A2 Antigen; D019073:HT29 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*
D029968:Escherichia coli Proteins; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli