A yeast surface display system for the discovery of ligands that trigger cell activation.
Opposing cells often communicate signalling events using multivalent interactions between receptors present on their cell surface. For example, T cells are typically activated when the T cell receptor (TCR) and its associated costimulatory molecules are multivalently engaged by the appropriate ligands present on an antigen presenting cell. In this report, yeast expressing high cell-surface levels of a TCR ligand (a recombinant antibody to the TCR Vbeta domain) were shown to act as 'pseudo' antigen presenting cells and induce T cell activation as monitored by increased levels of CD25 and CD69 and by downregulation of cell surface TCR. Similar levels of T cell activation could occur even when a 30-fold excess of irrelevant yeast was present, suggesting that such a yeast display system, by virtue of its ability to present ligands multivalently, may be used in highly sensitive procedures to identify novel polypeptides that interact multivalently with cell surface receptors and thereby trigger specific cellular responses.
Cho
B K
BK
Department of Biochemistry, University of Illinois, Urbana 61801, USA.
Kieke
M C
MC
Boder
E T
ET
Wittrup
K D
KD
Kranz
D M
DM
D000818:Animals; D017951:Antigen Presentation; D015703:Antigens, CD / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D015536:Down-Regulation; D005434:Flow Cytometry; D006825:Hybridomas / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D007840:Latex; D037181:Lectins, C-Type; D008024:Ligands; D008213:Lymphocyte Activation / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D008863:Microspheres; D010446:Peptide Fragments / Q000276:immunology* / Q000302:isolation & purification; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology*; D015375:Receptors, Interleukin-2 / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012441:Saccharomyces cerevisiae / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014162:Transfection; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D007840:Latex; D037181:Lectins, C-Type; D008024:Ligands; D010446:Peptide Fragments; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D000099192:CD69 Antigens; C055966:immunoglobulin Fv