Induction and Exhaustion of Lymphocytic Choriomeningitis Virus–specific Cytotoxic T Lymphocytes Visualized Using Soluble Tetrameric Major Histocompatibility Complex Class I–Peptide Complexes

• DOI: 10.1084/jem.187.9.1383
• Publication Date: 2002-07-26T16:49:30Z
• Authors: Awen Gallimore; Ann Glithero; Andrew Godkin; Alain C. Tissot; Andreas Plückthun; Tim Elliott; Hans Hengartner; Rolf Zinkernagel
• Keywords: Arenavirus

This study describes the construction of soluble major histocompatibility complexes consisting mouse class I molecule, H-2Db, chemically biotinylated β2 microglobulin and a peptide epitope derived from glycoprotein (GP; amino acids 33–41) lymphocytic choriomeningitis virus (LCMV). Tetrameric complexes, which were produced by mixing with phycoerythrin-labeled neutravidin, permitted direct analysis virus-specific cytotoxic T lymphocytes (CTLs) flow cytometry. technique was validated (a) staining CD8+ cells in spleens transgenic mice that express cell receptor (TCR) specific for H-2Db association GP33–41, (b) CTLs cerebrospinal fluid C57BL/6 (B6) had been infected intracranially LCMV-DOCILE. Staining spleen isolated B6 revealed up to 40% GP33 tetramer+ during initial phase LCMV infection. In contrast, tetramers did not stain 2 mo previously above background levels found naive mice. The fate analyzed acute infection challenged both intravenously high or low dose results show outcome is determined antigen load alone. Furthermore, data indicate deletion presence excessive preceded TCR downregulation dependent upon perforin.