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Isolation of CD4+ and CD8+ T-Cell Clones from Mice Immunized with Synthetic Peptides on Splenic Dendritic Cells
1Basel Institute for Immunology, Grenzacherstrasse 487, Basel, CH-4005, Switzerland
Splenic dendritic cells (DC) express high levels of MHC, co-stimulator, and adhesion molecules and have been shown to be extremely potent antigen-presenting cells for both CD4(+) and CD8(+) T-cell responses. Previous studies have shown that murine DC can be loaded with exogenous antigens and used to prime CD4(+), Class II-restricted T-cell responses in vivo. This article describes protocols for immunization using DC loaded with peptides bound to Class I or Class II molecules and for the derivation and characterization of CD4(+) and CD8(+) antigen-specific T-cell lines and clones. The rationale for using DC as antigen-presenting cells is discussed, together with the advantages and disadvantages of these cells compared with more conventional methods of immunization.
Splenic dendritic cells (DC) express high levels of MHC, co-stimulator, and adhesion molecules and have been shown to be extremely potent antigen-presenting cells for both CD4(+) and CD8(+) T-cell responses. Previous studies have shown that murine DC can be loaded with exogenous antigens and used to prime CD4(+), Class II-restricted T-cell responses in vivo. This article describes protocols for immunization using DC loaded with peptides bound to Class I or Class II molecules and for the derivation and characterization of CD4(+) and CD8(+) antigen-specific T-cell lines and clones. The rationale for using DC as antigen-presenting cells is discussed, together with the advantages and disadvantages of these cells compared with more conventional methods of immunization.
