Age-dependent alterations in the assembly of signal transduction complexesat the site of T cell/APC interaction

TCR interaction with peptide-MHC complexes triggers migration of protein ki nases, actin-binding proteins, and other accessory molecules to the T cell/ APC synapse. We used confocal immunofluorescence methods to show that the a dapter protein LAT (linker for activation of T cells) and the guanine nucle otide exchange factor Vav also move to the APC interface in mouse CD4 T cel ls conjugated to anti-CD3 hybridoma cells, and in TCR-transgenic CD4 cells conjugated to APC bearing agonist (but not closely related nonagonist) pept ides, The proportion of CD4(+) T cells able to relocalize LAT or Vav, or to relocate cytoplasmic NT-AT (NF-ATc) from cytoplasm to nucleus, declines ab out 2-fold in aged mice. The decline in LAT relocalization is accompanied b y a similar decline in tyrosine phosphorylation of LAT in CD4 cells stimula ted by CD3/CD4 cross-linking. Two-color experiments show that LAT redistrib ution is strongly associated with relocalization of both NF-ATc and protein kinase C-theta among individual cells, LAT migration to the immunological synapse depends on actin polymerization as well as on activity of Src famil y kinases, but aging leads to only a small change in the percentage of CD4 cells that redistribute P-actin to the site of APC contact. These results s uggest that defects in the ability of T cells from aged donors to move kina se substrates and coupling factors, including LAT and Vav, into the T cell/ APC contact region may contribute to the decline with age in NF-ATc-depende nt gene expression, and thus to defects in T cell clonal expansion.