title: Physical and functional bivalency observed among TCR/CD3 complexes isolated from primary T cells.
abstract: Unlike BCR and secreted Ig, TCR expression is not thought to occur in a bivalent form. The conventional monovalent model of TCR/CD3 is supported by published studies of complexes solubilized in the detergent digitonin, in which bivalency was not observed. We revisited the issue of TCR valency by examining complexes isolated from primary αβ T cells after solubilization in digitonin. Using immunoprecipitation followed by flow cytometry, we unexpectedly observed TCR/CD3 complexes that contained two TCRs per complex. Standard anti-TCR Abs, being bivalent themselves, tended to bind with double occupancy to bivalent TCRs; this property masked the presence of the second TCR per complex in certain Ab binding assays, which may partially explain why previous data did not reveal these bivalent complexes. We also found that the prevalence of bivalency among fully assembled, mature TCR/CD3 complexes was sufficient to impact the functional performance of immunoprecipitated TCRs in binding antigenic peptide/MHC-Ig fusion proteins. Both TCR positions per bivalent complex required an Ag-specific TCR to effect optimal binding to these soluble ligands. Therefore, we conclude that in primary T cells, TCR/CD3 complexes can be found that are physically and functionally bivalent. The expression of bivalent TCR/CD3 complexes has implications regarding potential mechanisms by which Ag may trigger signaling. It also suggests the possibility that the potential for bivalent expression could represent a general feature of Ag receptors.
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA. Schrum.Adam@mayo.edu
authors.forename: Adam G
authors.initials: AG
authors.lastname: Schrum
authors.forename: Diana
authors.initials: D
authors.lastname: Gil
authors.forename: Laurence A
authors.initials: LA
authors.lastname: Turka
authors.forename: Ed
authors.initials: E
authors.lastname: Palmer
mesh_terms: D056704:Adaptive Immunity / Q000235:genetics; D000818:Animals; D017252:CD3 Complex / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D005434:Flow Cytometry; D047468:Immunoprecipitation; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D055503:Protein Multimerization / Q000235:genetics / Q000276:immunology; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D016176:T-Lymphocyte Subsets / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism
chemical_list: D017252:CD3 Complex; D017260:Receptor-CD3 Complex, Antigen, T-Cell