tetano
Editor, Senior Moderator
J Immunol Methods. 2017 Sep 30. pii: S0022-1759(17)30426-X. doi: 10.1016/j.jim.2017.09.010. [Epub ahead of print]
[h=1]Multiplex peptide-MHC tetramer staining using mass cytometry for deep analysis of the influenza-specific T-cell response in mice.[/h] Fehlings M[SUP]1[/SUP], Chakarov S[SUP]2[/SUP], Simoni Y[SUP]2[/SUP], Shivshankar B[SUP]3[/SUP], Ginhoux F[SUP]2[/SUP], Newell EW[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Antigen-specific T cells play a crucial role for the host protective immunity against viruses and other diseases. The combination of mass cytometry together with a combinatorial multiplex tetramer staining has successfully been applied for probing and characterization of multiple antigen-specific CD8[SUP]+[/SUP] T cells in human blood samples. The present study shows that this approach can also be used to rapidly identify the magnitude of influenza-specific CD8[SUP]+[/SUP] T cell peptide dominance across lymph nodes and lungs in a murine model of a highly pathological influenza infection. Moreover, we also prove feasibility of this approach to be expandable for the concurrent identification of virus specific CD4[SUP]+[/SUP] T cells. By using a double coding approach, we probed for five influenza-specific MHCI-peptide complexes as well as one influenza-specific MHCII-peptide complex in the presence of irrelevant control peptides and show that this approach is capable of tracking antigen-specific T cells across individual lymph nodes and lungs. The simultaneous staining with 26 surface maker molecules further facilitated an in-depth characterization of T cells reacting with influenza epitopes and revealed tissue specific phenotypic differences between CD4[SUP]+[/SUP] T cells targeting the same pathogenic epitope. In conclusion, this approach provides the possibility for a rapid and comprehensive analysis of antigen-specific CD8[SUP]+[/SUP] and CD4[SUP]+[/SUP] T cells in different disease settings that might be advantageous for subsequent vaccine formulation strategies.
Copyright ? 2017. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Influenza; Mass cytometry; Multiplex combinatorial tetramer staining; T cell response
PMID: 28974367 DOI: 10.1016/j.jim.2017.09.010
[h=1]Multiplex peptide-MHC tetramer staining using mass cytometry for deep analysis of the influenza-specific T-cell response in mice.[/h] Fehlings M[SUP]1[/SUP], Chakarov S[SUP]2[/SUP], Simoni Y[SUP]2[/SUP], Shivshankar B[SUP]3[/SUP], Ginhoux F[SUP]2[/SUP], Newell EW[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Antigen-specific T cells play a crucial role for the host protective immunity against viruses and other diseases. The combination of mass cytometry together with a combinatorial multiplex tetramer staining has successfully been applied for probing and characterization of multiple antigen-specific CD8[SUP]+[/SUP] T cells in human blood samples. The present study shows that this approach can also be used to rapidly identify the magnitude of influenza-specific CD8[SUP]+[/SUP] T cell peptide dominance across lymph nodes and lungs in a murine model of a highly pathological influenza infection. Moreover, we also prove feasibility of this approach to be expandable for the concurrent identification of virus specific CD4[SUP]+[/SUP] T cells. By using a double coding approach, we probed for five influenza-specific MHCI-peptide complexes as well as one influenza-specific MHCII-peptide complex in the presence of irrelevant control peptides and show that this approach is capable of tracking antigen-specific T cells across individual lymph nodes and lungs. The simultaneous staining with 26 surface maker molecules further facilitated an in-depth characterization of T cells reacting with influenza epitopes and revealed tissue specific phenotypic differences between CD4[SUP]+[/SUP] T cells targeting the same pathogenic epitope. In conclusion, this approach provides the possibility for a rapid and comprehensive analysis of antigen-specific CD8[SUP]+[/SUP] and CD4[SUP]+[/SUP] T cells in different disease settings that might be advantageous for subsequent vaccine formulation strategies.
Copyright ? 2017. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Influenza; Mass cytometry; Multiplex combinatorial tetramer staining; T cell response
PMID: 28974367 DOI: 10.1016/j.jim.2017.09.010