T cells recognizing a 11mer influenza peptide complexed to H-2D(b) show promiscuity for peptide length.

T-cell repertoire is selected according to self peptide-MHC (major histocompatibility complex) complexes in the thymus. Although most peripheral T cells recognize specific pathogen-derived peptides complexed to self-MHC exclusively, some possess cross-reactivity to other self or foreign peptides presented by self-MHC molecules; a phenomenon often termed T-cell receptor (TCR) promiscuity or degeneracy. TCR promiscuity has been attributed to various autoimmune conditions. On the other hand, it is considered a mechanism for a relatively limited TCR repertoire to deal with a potentially much larger antigenic peptide repertoire. Such property has also been utilized to bypass self-tolerance for cancer vaccine development. Although many studies explored such degeneracy for peptide of the same length, few studies reported such properties for peptides of different length. In this study, we finely characterized the CD8(+) T-cell response specific for a 11mer peptide derived from influenza A viral polymerase basic protein 2. The short-term T-cell line, despite possessing highly biased TCR, was able to react with multiple peptides of different length sharing the same core sequence. Out data clearly showed the importance of detailed and quantitative assessments for such T-cell specificity. Our data also emphasize the importance of biochemical demonstration of the naturally presented minimal peptide.

Influenza A infection enhances cross-priming of CD8+ T cells to cell-associated antigens in a TLR7- and type I IFN-dependent fashion.

The initiation of antitumor immunity relies on dendritic cells (DCs) to cross-present cell-associated tumor Ag to CD8(+) T cells (T(CD8+)) due to a lack of costimulatory molecules on tumor cells. Innate danger signals have been demonstrated to enhance cross-priming of T(CD8+) to soluble as well as virally encoded Ags; however, their effect on enhancing T(CD8+) cross-priming to cell genome-encoded Ags remains unknown. Furthermore, influenza A virus (IAV) has not been shown to enhance antitumor immunity. Using influenza-infected allogeneic cell lines, we show in this study that T(CD8+) responses to cell-associated Ags can be dramatically enhanced due to enhanced T(CD8+) expansion. This enhanced cross-priming in part involves TLR7- but not TLR3-mediated sensing of IAV and is entirely dependent on MyD88 and IFN signaling pathways. We also showed that the inflammasome-induced IL-1 and IFN-γ did not play a role in enhancing cross-priming in our system. We further demonstrated in our ex vivo system that CD8(+) DCs are the only APCs able to prime TCR-transgenic T(CD8+). Importantly, plasmacytoid DCs and CD8(-) DCs were both able to enhance such priming when provided in coculture. These observations suggest that IAV infection of tumor cells may facilitate improved cross-presentation of tumor Ags and may be used to augment clinical vaccine efficacy.

A chimeric TCR-beta chain confers increased susceptibility to EAE.

Autoreactive myelin-specific CD4(+) T cells play an important role in CNS demyelination observed in MS and EAE. Consequently, it is important to understand the mechanisms of T cell receptor signalling leading to the activation of autoreactive T cells. We have previously generated a chimeric T cell receptor beta-chain (betaIII) displaying increased antigen sensitivity by exchanging most of the transmembrane and the intracellular domain of the TCR-beta chain with the corresponding TCR-gamma sequence. To investigate the effect of this "super-signalling" TCR in an autoimmune setting, we generated MOG(35-55) specific TCR transgenic mice expressing either the wild-type or the chimeric betaIII TCR-beta chain. We found that naïve transgenic T cells expressing the chimeric betaIII chain proliferated more extensively than wild-type cells in response to MOG(35-55)in vitro. Likewise, betaIII T cells skewed into a TH1 phenotype maintained the proliferative advantage over wild-type TH1 T cells at low antigen concentration. However, when skewed into a TH2 phenotype, there was no difference in proliferation between wild-type and betaIII T cells. Blocking of Fas-mediated cell death evenly affected wild-type and betaIII TH1 T cells and resulted in increased proliferation of both subsets, suggesting that betaIII T cells did not show defective Fas-FasL signalling. Finally, we found that betaIII TCR transgenic mice are more susceptible to EAE than wild-type TCR transgenic mice. We conclude that the change in the transmembrane domain of the TCR-beta chain affects TH1 T cells and the susceptibility to EAE, but does not affect TH2 cells. Investigating the molecular interaction within the TCR complex will help us to identify signalling pathways that can be manipulated to stop the progression of MS.