PMID- 22082029
OWN - NLM
STAT- MEDLINE
DCOM- 20120509
LR  - 20211021
IS  - 1479-5876 (Electronic)
IS  - 1479-5876 (Linking)
VI  - 9
DP  - 2011 Nov 14
TI  - Optimizing parameters for clinical-scale production of high IL-12 secreting 
      dendritic cells pulsed with oxidized whole tumor cell lysate.
PG  - 198
LID - 10.1186/1479-5876-9-198 [doi]
AB  - BACKGROUND: Dendritic cells (DCs) are the most potent antigen-presenting cell 
      population for activating tumor-specific T cells. Due to the wide range of 
      methods for generating DCs, there is no common protocol or defined set of 
      criteria to validate the immunogenicity and function of DC vaccines. METHODS: 
      Monocyte-derived DCs were generated during 4 days of culture with recombinant 
      granulocyte-macrophage colony stimulating factor and interleukin-4, and pulsed 
      with tumor lysate produced by hypochlorous acid oxidation of tumor cells. 
      Different culture parameters for clinical-scale DC preparation were investigated, 
      including: 1) culture media; 2) culture surface; 3) duration of activating DCs 
      with lipopolysaccharide (LPS) and interferon (IFN)-gamma; 4) method of DC 
      harvest; and 5) cryomedia and final DC product formulation. RESULTS: DCs cultured 
      in CellGenix DC media containing 2% human AB serum expressed higher levels of 
      maturation markers following lysate-loading and maturation compared to culturing 
      with serum-free CellGenix DC media or AIM-V media, or 2% AB serum supplemented 
      AIM-V media. NunclonDelta surface, but not Corning((R)) tissue-culture treated surface 
      and Corning((R)) ultra-low attachment surface, were suitable for generating an 
      optimal DC phenotype. Recombinant trypsin resulted in reduced major 
      histocompatibility complex (MHC) Class I and II expression on mature 
      lysate-loaded DCs, however presentation of MHC Class I peptides by DCs was not 
      impaired and cell viability was higher compared to cell scraping. Preservation of 
      DCs with an infusible cryomedia containing Plasma-Lyte A, dextrose, sodium 
      chloride injection, human serum albumin, and DMSO yielded higher cell viability 
      compared to using human AB serum containing 10% DMSO. Finally, activating DCs for 
      16 hours with LPS and IFN-gamma stimulated robust mixed leukocyte reactions (MLRs), 
      and high IL-12p70 production in vitro that continued for 24 hours after the 
      cryopreserved DCs were thawed and replated in fresh media. CONCLUSIONS: This 
      study examined criteria including DC phenotype, viability, IL-12p70 production 
      and the ability to stimulate MLR as metrics of whole oxidized tumor lysate-pulsed 
      DC immunogenicity and functionality. Development and optimization of this unique 
      method is now being tested in a clinical trial of autologous oxidized tumor 
      lysate-pulsed DC in clinical-scale in recurrent ovarian, primary peritoneal or 
      fallopian tube cancer (NCT01132014).
FAU - Chiang, Cheryl L-L
AU  - Chiang CL
AD  - Ovarian Cancer Research Center, University of Pennsylvania, Philadelphia, 19104, 
      USA.
FAU - Maier, Dawn A
AU  - Maier DA
FAU - Kandalaft, Lana E
AU  - Kandalaft LE
FAU - Brennan, Andrea L
AU  - Brennan AL
FAU - Lanitis, Evripidis
AU  - Lanitis E
FAU - Ye, Qunrui
AU  - Ye Q
FAU - Levine, Bruce L
AU  - Levine BL
FAU - Czerniecki, Brian J
AU  - Czerniecki BJ
FAU - Powell, Daniel J Jr
AU  - Powell DJ Jr
FAU - Coukos, George
AU  - Coukos G
LA  - eng
SI  - ClinicalTrials.gov/NCT01132014
GR  - P50-CA083638/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20111114
PL  - England
TA  - J Transl Med
JT  - Journal of translational medicine
JID - 101190741
RN  - 0 (Cell Extracts)
RN  - 0 (Culture Media)
RN  - 187348-17-0 (Interleukin-12)
RN  - 712K4CDC10 (Hypochlorous Acid)
RN  - EC 3.4.21.4 (Trypsin)
SB  - IM
MH  - Cell Culture Techniques/*methods
MH  - Cell Differentiation/drug effects
MH  - Cell Extracts/*pharmacology
MH  - Cell Line, Tumor
MH  - Cell Survival/drug effects
MH  - Cryopreservation
MH  - Culture Media/pharmacology
MH  - Dendritic Cells/*cytology/drug effects/*metabolism
MH  - Humans
MH  - Hypochlorous Acid/pharmacology
MH  - Interleukin-12/biosynthesis/*metabolism
MH  - Lymphocyte Culture Test, Mixed
MH  - Oxidation-Reduction/drug effects
MH  - Phenotype
MH  - Time Factors
MH  - Trypsin/metabolism
PMC - PMC3283529
EDAT- 2011/11/16 06:00
MHDA- 2012/05/10 06:00
PMCR- 2011/11/14
CRDT- 2011/11/16 06:00
PHST- 2011/06/27 00:00 [received]
PHST- 2011/11/14 00:00 [accepted]
PHST- 2011/11/16 06:00 [entrez]
PHST- 2011/11/16 06:00 [pubmed]
PHST- 2012/05/10 06:00 [medline]
PHST- 2011/11/14 00:00 [pmc-release]
AID - 1479-5876-9-198 [pii]
AID - 10.1186/1479-5876-9-198 [doi]
PST - epublish
SO  - J Transl Med. 2011 Nov 14;9:198. doi: 10.1186/1479-5876-9-198.