Activation of a cytotoxic T-cell is a complex multistep process, and tools to study the molecular events and their dynamics that result in T-cell activation in situ and in vivo are scarce. Here, we... Show moreActivation of a cytotoxic T-cell is a complex multistep process, and tools to study the molecular events and their dynamics that result in T-cell activation in situ and in vivo are scarce. Here, we report the design and use of conditional epitopes for time-controlled T-cell activation in vivo. We show that trans-cyclooctene-protected SIINFEKL (with the lysine amine masked) is unable to elicit the T-cell response characteristic for the free SIINFEKL epitope. Epitope uncaging by means of an inverse-electron demand Diels–Alder (IEDDA) event restored T-cell activation and provided temporal control of T-cell proliferation in vivo. Show less
The aim of this thesis is to explore the use of Bioorthogonal Antigens to study the cross-presentation pathway. Bioorthogonal Antigens are antigens carrying bioorthogonal groups in specific... Show moreThe aim of this thesis is to explore the use of Bioorthogonal Antigens to study the cross-presentation pathway. Bioorthogonal Antigens are antigens carrying bioorthogonal groups in specific amino acid positions within the epitope region that can be reacted selectively within/on the cell using bioorthogonal ligation strategies. Incorporation of bioorthogonal groups into antigens has an advantage over other methods because most of the groups are stable to proteolysis and are small enough to have a minimal impact on routing and loading onto MHC-I molecules. In the future, these antigens have potential to be applied for imaging of the entire cross-presentation pathway using a single bioorthogonal handle. Show less
