PMID- 23653155
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20130509
LR  - 20220809
IS  - 2162-2531 (Print)
IS  - 2162-2531 (Electronic)
IS  - 2162-2531 (Linking)
VI  - 2
IP  - 5
DP  - 2013 May 7
TI  - Evaluation of RNA Amplification Methods to Improve DC Immunotherapy Antigen 
      Presentation and Immune Response.
PG  - e91
LID - 10.1038/mtna.2013.18 [doi]
AB  - 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 result in amplification of longer transcripts, greater translational 
      competence, and a higher fidelity representation of the starting total RNA 
      population. To study the properties of amplified RNA after transfection into 
      human DCs, we measured protein expression levels of defined antigens coamplified 
      with the starting total RNA populations and measured antigen-specific T cell 
      expansion in autologous DC-T cell co-cultured in vitro. We conclude from these 
      analyses that the improved RNA amplification process results in superior protein 
      expression levels and a greater capacity of the transfected DCs to induce 
      multifunctional antigen-specific memory T cells.Molecular Therapy-Nucleic Acids 
      (2013) 2, e91; doi:10.1038/mtna.2013.18; published online 7 May 2013.
FAU - Slagter-Jager, Jacoba G
AU  - Slagter-Jager JG
AD  - Argos Therapeutics, Durham, North Carolina, USA.
FAU - Raney, Alexa
AU  - Raney A
FAU - Lewis, Whitney E
AU  - Lewis WE
FAU - Debenedette, Mark A
AU  - Debenedette MA
FAU - Nicolette, Charles A
AU  - Nicolette CA
FAU - Tcherepanova, Irina Y
AU  - Tcherepanova IY
LA  - eng
PT  - Journal Article
DEP - 20130507
PL  - United States
TA  - Mol Ther Nucleic Acids
JT  - Molecular therapy. Nucleic acids
JID - 101581621
PMC - PMC4817939
EDAT- 2013/05/09 06:00
MHDA- 2013/05/09 06:01
PMCR- 2013/05/01
CRDT- 2013/05/09 06:00
PHST- 2013/05/09 06:00 [entrez]
PHST- 2013/05/09 06:00 [pubmed]
PHST- 2013/05/09 06:01 [medline]
PHST- 2013/05/01 00:00 [pmc-release]
AID - S2162-2531(16)30150-0 [pii]
AID - 10.1038/mtna.2013.18 [doi]
PST - epublish
SO  - Mol Ther Nucleic Acids. 2013 May 7;2(5):e91. doi: 10.1038/mtna.2013.18.