Vaccine T-cell epitope selection by a peptide competition assay.

Abstract

The binding of several peptides derived from the Plasmodium falkiparum circumsporozoite protein (CS protein) to the human major histocompatibility complex class II proteins HLA-DR5 and -DRw6 was examined in a competition assay. Fixed antigen-presenting cells (APCs) were incubated with various concentrations of each peptide and suboptimal concentrations of stimulator peptides. The binding of the CS peptides to DR5 or DRw6 proteins was then determined in a proliferation assay using two established DR5 or DRw6-restricted T-cell clones with specificity for the stimulator peptides as responder cells. One offive CS peptides, comprising together about 50% of the CS protein sequence, was found to compete with the binding of the stimulator peptides to DR5 and DRw6. The CS peptide CS-(378-398), binding to DR5 and DRw6, was then shown to be able to induce primary in vitro responses of T cells from donors with DR5 and DRw6 haplotypes. CS-(378-398)-induced T-cell clones responded not only to the homologous peptide but also to the native CS protein in the presence of appropriate APCs. The strategy we have applied is of considerable general interest for the engineering of vaccines against any pathogen, since it greatly facilitates the selection of appropriate T-cell epitopes to be incorporated in the vaccine.

Gold-specific T cells in rheumatoid arthritis patients treated with gold.

“…Competition for antigen presentation has been described elsewhere (20). Briefly, autologous EBV-B cells were fixed by resuspending them in 0.025% glutaraldehyde for 90 s. …”

Induction of antigen‐specific CD8+ cytolytic T cells by the exogenous bacterial antigen streptolysin O in rhesus monkeys

“…5~ 105). In some experiments, APC were glutaraldehyde fixed following a slightly modified version of the described protocol [22]. Before or after incubation at 37°C for 4 h with SLO, APC were extensively washed and were fixed with 0.05% glutaraldehyde (Sigma) in PBS for 90 s. The reaction was then stopped by adding 5 vol of 0.…”

Transcatheter closure of a ruptured ventricular septum following inferior myocardial infarction and cardiogenic shock

“…Trop‐2 is highly homologous to the cell–cell adhesion molecule Trop‐1 (17‐1A, Ep‐CAM, GA733‐2) 23, 24, 25, 26. The latter has been utilized as a target for antibody immunotherapy,27, 28, 29, 30, 31, 32, 33, 34 and anti‐Trop‐1 antibodies can cause a significant reduction in mortality and relapse rates of resected colon cancer patients 33. Cellular anti‐Trop‐1 responses have been demonstrated in tumor patients35 and anti‐Trop‐1 CTL are induced efficiently by peptide‐pulsed APC 36.…”