Evaluation of RNA Amplification Methods to Improve DC Immunotherapy Antigen Presentation and Immune Response.

ABSTRACT: Dendritic cells (DCs) transfected with total amplified tumor cell RNA have the potential to induce broad antitumor immune responses. However, analytical methods required for quantitatively assessing the integrity, fidelity, and functionality of the amplified RNA are lacking. We have developed a series of assays including gel electrophoresis, northern blot, capping efficiency, and microarray analysis to determine integrity and fidelity and a model system to assess functionality after transfection into human DCs. We employed these tools to demonstrate that modifications to our previously reported total cellular RNA amplification process including the use of the Fast Start High Fidelity (FSHF) PCR enzyme, T7 Powerswitch primer, post-transcriptional capping and incorporation of a type 1 cap re

SUBMITTER: Slagter-Jager JG 

PROVIDER: S-EPMC4817939 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature