PMID- 10986562
OWN - NLM
STAT- MEDLINE
DCOM- 20001026
LR  - 20231213
IS  - 1043-0342 (Print)
IS  - 1043-0342 (Linking)
VI  - 11
IP  - 13
DP  - 2000 Sep 1
TI  - Efficient gene transfer to human peripheral blood monocyte-derived dendritic 
      cells using human immunodeficiency virus type 1-based lentiviral vectors.
PG  - 1901-9
AB  - Dendritic cells (DCs) are potent antigen-presenting cells and are capable of 
      activating naive T cells. Gene transfer of tumor antigen and cytokine genes into 
      DCs could be an important strategy for immunotherapeutic applications. Dendritic 
      cells derived from peripheral blood monocytes do not divide and are therefore 
      poor candidates for gene transfer by Moloney murine leukemia virus 
      (Mo-MuLV)-based retroviral vectors. Lentiviral vectors are emerging as a powerful 
      tool for gene delivery into dividing and nondividing cells. A three-plasmid 
      expression system pseudotyped with the envelope from vesicular stomatitis virus 
      (VSV-G) was used to generate lentiviral vector particles expressing enhanced 
      green fluorescent protein (EGFP). Peripheral blood monocyte-derived DCs were 
      cultured in the presence of GM-CSF and IL-4 and transduced with lentiviral or 
      Mo-MuLV-based vectors expressing EGFP. FACS analysis of lentiviral 
      vector-transduced DCs derived either from normal healthy volunteers or from 
      melanoma patients demonstrated transduction efficiency ranging from 70 to 90% 
      compared with 2-8% using Mo-MuLV-based vectors pseudotyped with VSV-G. Comparison 
      of lentiviral vectors expressing EGFP driven by CMV or human PGK promoters showed 
      similar levels of transgene expression. Lentiviral vector preparations produced 
      in the absence of HIV accessory proteins transduced DCs at efficiencies equal to 
      vectors produced with accessory proteins. Alu-HIV-1 LTR PCR demonstrated the 
      genomic integration of the lentiviral vector in the transduced DCs. Transduced 
      cells showed characteristic dendritic cell phenotype and strong allostimulatory 
      capacity and maintained the ability to respond to activation signals such as CD40 
      ligand and lipopolysaccharide. These results provide evidence that lentiviral 
      vectors are efficient tools for gene transfer and expression in monocyte-derived 
      DCs that could be useful for immunotherapeutic applications.
FAU - Chinnasamy, N
AU  - Chinnasamy N
AD  - Clinical Gene Therapy Branch, NHGRI, Bethesda, MD 20892, USA.
FAU - Chinnasamy, D
AU  - Chinnasamy D
FAU - Toso, J F
AU  - Toso JF
FAU - Lapointe, R
AU  - Lapointe R
FAU - Candotti, F
AU  - Candotti F
FAU - Morgan, R A
AU  - Morgan RA
FAU - Hwu, P
AU  - Hwu P
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Hum Gene Ther
JT  - Human gene therapy
JID - 9008950
RN  - 0 (Antigens, CD)
RN  - 0 (Immunoglobulins)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Viral Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 187348-17-0 (Interleukin-12)
SB  - IM
MH  - Alu Elements/genetics
MH  - Antigens, CD
MH  - Cytomegalovirus/genetics
MH  - Dendritic Cells/immunology/*physiology/virology
MH  - *Gene Transfer Techniques
MH  - Green Fluorescent Proteins
MH  - HIV-1/genetics
MH  - Humans
MH  - Immunoglobulins/metabolism
MH  - Interleukin-12/metabolism
MH  - Lentivirus/*genetics
MH  - Luminescent Proteins/genetics/metabolism
MH  - Melanoma/genetics/pathology
MH  - Membrane Glycoproteins/metabolism
MH  - Moloney murine leukemia virus/genetics
MH  - Monocytes/*cytology/virology
MH  - Promoter Regions, Genetic
MH  - T-Lymphocytes/immunology
MH  - Vesicular stomatitis Indiana virus/genetics
MH  - Viral Proteins/genetics
MH  - CD83 Antigen
EDAT- 2000/09/15 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/09/15 11:00
PHST- 2000/09/15 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/09/15 11:00 [entrez]
AID - 10.1089/10430340050129512 [doi]
PST - ppublish
SO  - Hum Gene Ther. 2000 Sep 1;11(13):1901-9. doi: 10.1089/10430340050129512.