title: Efficient and sustained gene expression in primary T lymphocytes and primary and cultured tumor cells mediated by adeno-associated virus plasmid DNA complexed to cationic liposomes.
abstract: We have used cationic liposomes to facilitate adeno-associated virus (AAV) plasmid transfections of primary and cultured cell types. AAV plasmid DNA complexed with liposomes showed levels of expression several fold higher than those of complexes with standard plasmids. In addition, long-term expression (> 30 days) of the gene, unlike the transient expression demonstrated by typical liposome-mediated transfection with standard plasmids, was observed. Southern analysis of chromosomal DNA further substantiated the hypothesis that the long-term expression was due to the presence of the transgene in the AAV plasmid-transfected group and not in the standard plasmid-transfected group. AAV plasmid-liposome complexes induced levels of transgene expression comparable to those obtained by recombinant AAV transduction. Primary breast, ovarian, and lung tumor cells were transfectable with the AAV plasmid DNA-liposome complexes. Transfected primary and cultured tumor cells were able to express transgene product even after lethal irradiation. High-level gene expression was also observed in freshly isolated CD3+, CD4+, and CD8+ T cells from normal human peripheral blood. Transfection efficiency ranged from 10 to 50% as assessed by intracellular interleukin-2 levels in interleukin-2-transfected cells. The ability to express transgenes in primary tumor and lymphoid cells may be applied toward tumor vaccine studies and protocols which may eventually permit highly specific modulation of the cellular immune response in cancer and AIDS.
authors.affiliation: Applied Immune Sciences, Inc., Santa Clara, California 95054.
authors.forename: R
authors.initials: R
authors.lastname: Philip
authors.forename: E
authors.initials: E
authors.lastname: Brunette
authors.forename: L
authors.initials: L
authors.lastname: Kilinski
authors.forename: D
authors.initials: D
authors.lastname: Murugesh
authors.forename: M A
authors.initials: MA
authors.lastname: McNally
authors.forename: K
authors.initials: K
authors.lastname: Ucar
authors.forename: J
authors.initials: J
authors.lastname: Rosenblatt
authors.forename: T B
authors.initials: TB
authors.lastname: Okarma
authors.forename: J S
authors.initials: JS
authors.lastname: Lebkowski
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D000818:Animals; D015139:Blotting, Southern; D001943:Breast Neoplasms / Q000378:metabolism*; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000096:biosynthesis; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D000229:Dependovirus / Q000235:genetics*; D004337:Drug Carriers; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D008081:Liposomes; D008175:Lung Neoplasms / Q000378:metabolism*; D008297:Male; D010051:Ovarian Neoplasms / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage*; D011471:Prostatic Neoplasms; D051381:Rats; D016176:T-Lymphocyte Subsets / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms
chemical_list: D004337:Drug Carriers; D007376:Interleukin-2; D008081:Liposomes; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase