Using a baculovirus display library to identify MHC class I mimotopes.
We have developed a baculovirus-based display system for identifying antigen mimotopes for MHC class I-specific T cells. The mouse MHC class I molecule, Dd, was displayed on baculovirus-infected insect cells with a library of 9- and 10-mer peptides tethered via a flexible linker to the N terminus of beta2 microglobulin. As a test case, the library was screened by flow cytometry by using a multimeric fluorescent alphabetaTCR from a mouse T cell specific for Dd plus an unknown self peptide. A mimotope was identified that, when bound to Dd, stimulated the T cell to secret IL-2. The sequence of the mimotope was used to identify a self peptide present in a mouse protein, Spin. The Spin peptide, when complexed with Dd, also activated the T cell. This technique should be generally useful in identifying and manipulating MHC class I peptide mimotopes and epitopes.
Wang
Yibing
Y
Howard Hughes Medical Institute, Integrated Department of Immunology, National Jewish Medical and Research Center, 1400 Jackson Street, Denver, CO 80206, USA.
Rubtsov
Anatoyla
A
Heiser
Ryan
R
White
Janice
J
Crawford
Frances
F
Marrack
Philippa
P
Kappler
John W
JW
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics; D002460:Cell Line; D004274:DNA, Recombinant / Q000737:chemistry / Q000235:genetics; D005434:Flow Cytometry; D015395:Histocompatibility Antigens Class I / Q000737:chemistry* / Q000235:genetics*; D007376:Interleukin-2 / Q000096:biosynthesis; D051379:Mice; D008869:Microtubule-Associated Proteins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D010750:Phosphoproteins / Q000235:genetics; D011487:Protein Conformation; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology
D018797:Cell Cycle Proteins; D004274:DNA, Recombinant; D015395:Histocompatibility Antigens Class I; D007376:Interleukin-2; D008869:Microtubule-Associated Proteins; D019151:Peptide Library; D010750:Phosphoproteins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; C104434:spindlin