Mucin gene (MUC1) transfer into human dendritic cells by cationic liposomes and recombinant adenovirus.
BACKGROUND
Dendritic cells (DC) as antigen presenting cells play an important role in immunotherapy of cancer. Mucin, encoded by the gene MUC1, is a human tumor antigen expressed in breast, pancreatic and ovarian cancers. Therefore, MUC1-transfected DC would be an attractive tool in constructing cancer vaccines.


METHODS
Using two different cationic liposome preparations and, for comparison, a recombinant adenovirus expressing mucin, we tested the efficiency of mucin gene transfer into DC by flow cytometry. We investigated if these transfected DC were able to specifically stimulate autologous peripheral blood lymphocytes (PBL) from healthy donors.


RESULTS
Flow cytometry revealed that 5-20% of DC transfected with liposomes Lipofectin and 20-40% of DC transduced with adenovirus expressed the relevant mucin epitopes. The expression of mucin on DC was similar to the expression of mucin found on carcinoma cells. After antigen uptake, DC specifically stimulated autologous PBL.


CONCLUSIONS
We have shown that cationic liposomal gene transfer into human DC was feasible. We could obtain antigen specific stimulation of PBL at a similar rate as with adenoviral MUC1-transduced DC.
Pecher
G
G
Charité, Humboldt-University Berlin, AG Molecular Gene- and Immunotherapy, Germany. gpecher@mdc-berlin.de
Spahn
G
G
Schirrmann
T
T
Kulbe
H
H
Ziegner
M
M
Schenk
J A
JA
Sandig
V
V
D000256:Adenoviridae / Q000235:genetics; D015703:Antigens, CD / Q000096:biosynthesis; D018949:Antigens, CD1 / Q000096:biosynthesis; D018122:B7-1 Antigen / Q000096:biosynthesis; D051940:B7-2 Antigen; D002411:Cation Exchange Resins; D002412:Cations; D018076:DNA, Complementary / Q000008:administration & dosage / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000096:biosynthesis; D006801:Humans; D008055:Lipids; D008081:Liposomes; D008213:Lymphocyte Activation / Q000276:immunology; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D018396:Mucin-1 / Q000235:genetics* / Q000276:immunology; D010714:Phosphatidylethanolamines; D010835:Phytohemagglutinins / Q000494:pharmacology; D014162:Transfection / Q000379:methods*
D015703:Antigens, CD; D018949:Antigens, CD1; D018122:B7-1 Antigen; D051940:B7-2 Antigen; C419393:CD1a antigen; C497964:CD86 protein, human; D002411:Cation Exchange Resins; D002412:Cations; D018076:DNA, Complementary; D006684:HLA-DR Antigens; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes; D008562:Membrane Glycoproteins; D018396:Mucin-1; D010714:Phosphatidylethanolamines; D010835:Phytohemagglutinins; C020888:1,2-dielaidoylphosphatidylethanolamine