HIV molecular immunology database
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|Immunogen||in vitro stimulation or selection|
Leggatt1997 G. R. Leggatt, M. A. Alexander-Miller, A. Kumar, S. L. Hoffman, and J. A. Berzofsky. Cytotoxic T Lymphocyte (CTL) Adherence Assay (CAA): A Non-Radioactive Assay for Murine CTL Recognition of Peptide-MHC Class I Complexes. J. Immunol. Methods, 201:1-10, 1997. This paper describes a novel assay, the CTL adhesion assay (CAA), and uses an HIV epitope in a murine system as a model system. CAA is a rapid, simple screening method for identifying cytolytic epitopes for a given CTL line, and may also identify peptides that cause T cell activation and adherence but not cytolytic activity. Cytotoxic T lymphocytes (CTL) form an important immune surveillance system against intracellular pathogens. Here we describe a simple, visual assay for identifying peptides specifically recognized by CTL, based on the discovery that CTL develop increased adhesive properties upon TCR triggering. Several CTL lines were shown to pellet to the bottom of a round bottom 96-well plate in the absence of peptide. In contrast, these same CTL lines incubated with their cognate peptide, allowing them to present peptide to each other, adhered to the sides of the well and were readily distinguished by macroscopic visual examination of the plate after 4-5 h or overnight incubation. This CTL adherence assay (CAA) demonstrated peptide specificity and MHC restriction, and was titratable with peptide concentration. With this technique, a minimal-sized, malaria CTL epitope was correctly identified from a panel of overlapping nonamers, although the adherence pattern of two mono-substituted, variant peptides was less. PubMed ID: 9053406. Show all entries for this paper.