title: Discrepancy between ELISPOT IFN-gamma secretion and binding of A2/peptide multimers to TCR reveals interclonal dissociation of CTL effector function from TCR-peptide/MHC complexes half-life.
abstract: Activation of CD8(+) cytolytic T lymphocytes (CTLs) by antigen is triggered by the interaction of clonotypic alphabeta T cell receptors (TCRs) with antigenic peptides bound to MHC class I molecules (pMHC complexes). Fluorescent multimeric pMHC complexes have been shown to specifically stain antigen-specific CTLs by directly binding the TCR. In tumor-infiltrating lymphocytes from a melanoma patient we found a high frequency of tyrosinase(368-376) peptide-specific cells as detected by IFN-gamma ELISPOT, without detectable staining with the corresponding A2/peptide multimers. Surprisingly, these T cells were able to lyse tyrosinase(368-376) peptide-pulsed target cells as efficiently as other specific T cells that were stained by multimers. Analysis of the staining patterns under different conditions of incubation time and temperature revealed that these results were explained by major differences in TCR-multimeric ligand interaction kinetics among the clones. Whereas no direct quantitative correlation between antigenic peptide concentration required for CTL effector functions and equilibrium multimer binding was observed interclonally, the latter was profoundly affected by the kinetics of TCR-ligand interaction. More importantly, our data indicate that similar levels of T cell activation can be achieved by independent CD8(+) T cell clonotypes displaying different TCR/pMHC complex dissociation rates.
authors.affiliation: Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, and Multidisciplinary Oncology Center, University Hospital, 1011 Lausanne, Switzerland.
authors.forename: V
authors.initials: V
authors.lastname: Rubio-Godoy
authors.forename: V
authors.initials: V
authors.lastname: Dutoit
authors.forename: D
authors.initials: D
authors.lastname: Rimoldi
authors.forename: D
authors.initials: D
authors.lastname: Lienard
authors.forename: F
authors.initials: F
authors.lastname: Lejeune
authors.forename: D
authors.initials: D
authors.lastname: Speiser
authors.forename: P
authors.initials: P
authors.lastname: Guillaume
authors.forename: J C
authors.initials: JC
authors.lastname: Cerottini
authors.forename: P
authors.initials: P
authors.lastname: Romero
authors.forename: D
authors.initials: D
authors.lastname: Valmori
mesh_terms: D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D015789:HLA-A2 Antigen / Q000737:chemistry / Q000378:metabolism*; D006207:Half-Life; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology; D046911:Macromolecular Substances; D008545:Melanoma / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D015789:HLA-A2 Antigen; D008024:Ligands; D046911:Macromolecular Substances; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma