Hi everyone,
Now I am generating CTLs using dendritic cells pulsed specific peptide, according to many protocols and papers, they have generated CTLs for nearly 1 month.
And now I am trying to reduce the time for CTLs generation (as short as I can) using dendritic cells pulsed specific peptide.
So, please help me to solve this issue.
Thank you sincerely!
Dear Cuong Manh Vo, the most probably, simple pulsing of DC by peptide results in inappropriate processing of the antigen. It is well known that externally trapped proteins and peptides successfully rich endosomal processing pathway and are presented by MHC class II molecules capable to stimulate CD4 T cells (not CD8 CTLs). For induction CTLs it would be preferable to use APC with endogenous expression of protein, containing peptide of interest. In this case, antigenic protein will rich proteasomal processing pathway with subsequent transport into EPR and loading onto MHC class I molecules capable to stimulate CD8 CTLs. Second important point is that you can use T lymphocytes from previously immunized organism.
Dear Dmitry B. Kaxansky, thank you so much for your help, your information is so important for me. recently I am using DCs pulsed tumor antigen for cancer treatment. for clinical application, we have to use autologous naive T cells, so that we can not use T lymphocytes from previously immunized organism.
Dear Cuong Manh Vo,
the following publication offers a lot of Information for CTL Generation an dsholud answer your questions:
Wölfl, M., Greenberg, P.D., 2014. Antigen-specific activation and cytokine-facilitated expansion of naive, human CD8+ T cells. Nature Protocols 9, 950–966. doi:10.1038/nprot.2014.064.
Good luck