Functional Similarity and Differences Between Selection-Independent CD4−CD8− αβ T Cells and Positively Selected CD8 T Cells Expressing the Same TCR and the Induction of Anergy in CD4−CD8− αβ T Cells in Antigen-Expressing Mice

In TCR-αβ transgenic mice, CD4−CD8− TCR-αβ+ (αβ DN) cells arise in the absence of positively selecting MHC molecules and are resistant to clonal deletion in Ag-expressing mice. In this study the activation requirements and functional properties of αβ double-negative (DN) cells were compared with those of positively selected CD8+ cells expressing equivalent levels of the same MHC class I-restricted transgenic TCR. We found that positively selected CD8+ cells required a lower density of the antigenic ligand for optimal proliferative responses compared with αβ DN cells derived from nonpositively selecting mice. However, when the CD8 coreceptor on CD8+ cells was blocked with an anti-CD8 mAb, both αβ DN and CD8+ cells exhibited the same dose-response curve to the antigenic ligand and the same dependence on CD28/B7 costimulation. Positively selected CD8+ cells also differed from αβ DN cells in that they differentiated into more efficient killers and IL-2 producers after Ag stimulation, even after CD8 blockade. However, Ag-activated αβ DN and CD8+ cells were equally efficient in producing IFN-γ, suggesting that this functional property is independent of positive selection. We also found that αβ DN cells recovered from the lymph nodes of Ag-expressing mice were functionally anergic. This anergic state was associated with defective proliferation and IL-2 production in response to Ag stimulation. These observations indicate that αβ DN cells can be anergized in vivo by physiological levels of the antigenic ligand.