Induction and exhaustion of lymphocytic choriomeningitis virus-specific cytotoxic T lymphocytes visualized using soluble tetrameric major histocompatibility complex class I-peptide complexes.
This study describes the construction of soluble major histocompatibility complexes consisting of the mouse class I molecule, H-2Db, chemically biotinylated beta2 microglobulin and a peptide epitope derived from the glycoprotein (GP; amino acids 33-41) of lymphocytic choriomeningitis virus (LCMV). Tetrameric class I complexes, which were produced by mixing the class I complexes with phycoerythrin-labeled neutravidin, permitted direct analysis of virus-specific cytotoxic T lymphocytes (CTLs) by flow cytometry. This technique was validated by (a) staining CD8+ cells in the spleens of transgenic mice that express a T cell receptor (TCR) specific for H-2Db in association with peptide GP33-41, and (b) by staining virus-specific CTLs in the cerebrospinal fluid of C57BL/6 (B6) mice that had been infected intracranially with LCMV-DOCILE. Staining of spleen cells isolated from B6 mice revealed that up to 40% of CD8(+) T cells were GP33 tetramer+ during the initial phase of LCMV infection. In contrast, GP33 tetramers did not stain CD8+ T cells isolated from the spleens of B6 mice that had been infected 2 mo previously with LCMV above the background levels found in naive mice. The fate of virus-specific CTLs was analyzed during the acute phase of infection in mice challenged both intracranially and intravenously with a high or low dose of LCMV-DOCILE. The results of the study show that the outcome of infection by LCMV is determined by antigen load alone. Furthermore, the data indicate that deletion of virus-specific CTLs in the presence of excessive antigen is preceded by TCR downregulation and is dependent upon perforin.
Gallimore
A
A
Institute of Experimental Immunology, CH-8091, Zürich, Switzerland. awen.gallimore@ndm.ox.ac.uk
Glithero
A
A
Godkin
A
A
Tissot
A C
AC
Plückthun
A
A
Elliott
T
T
Hengartner
H
H
Zinkernagel
R
R
D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D005434:Flow Cytometry; D006023:Glycoproteins / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D007371:Interferon-gamma / Q000032:analysis; D008217:Lymphocytic choriomeningitis virus / Q000737:chemistry / Q000276:immunology*; D008285:Major Histocompatibility Complex / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000172:deficiency; D051379:Mice; D008822:Mice, Transgenic; D010455:Peptides / Q000276:immunology; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D011487:Protein Conformation; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D013154:Spleen / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000821:virology*; D001613:beta 2-Microglobulin / Q000378:metabolism
D006023:Glycoproteins; D015395:Histocompatibility Antigens Class I; D008562:Membrane Glycoproteins; D010455:Peptides; D052899:Pore Forming Cytotoxic Proteins; D011948:Receptors, Antigen, T-Cell; D001613:beta 2-Microglobulin; D054353:Perforin; D007371:Interferon-gamma