       Document 0072
 DOCN  M9460072
 TI    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.
 DT    9404
 AU    Philip R; Brunette E; Kilinski L; Murugesh D; McNally MA; Ucar K;
       Rosenblatt J; Okarma TB; Lebkowski JS; Applied Immune Sciences, Inc.,
       Santa Clara, California 95054.
 SO    Mol Cell Biol. 1994 Apr;14(4):2411-8. Unique Identifier : AIDSLINE
       MED/94187712
 AB    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.
 DE    Acquired Immunodeficiency Syndrome/IMMUNOLOGY  Animal  Bladder Neoplasms
       Blotting, Southern  Breast Neoplasms/*METABOLISM  Cells, Cultured
       Chloramphenicol Acetyltransferase/ANALYSIS/BIOSYNTHESIS
       Dependovirus/*GENETICS  Drug Carriers  DNA/ADMINISTRATION &
       DOSAGE/*GENETICS  Female  *Gene Expression  Genetic Vectors  Human
       Interleukin-2/*BIOSYNTHESIS/GENETICS  Liposomes  Lung
       Neoplasms/*METABOLISM  Male  Ovarian Neoplasms/*METABOLISM
       Plasmids/*ADMINISTRATION & DOSAGE  Prostatic Neoplasms  Rats
       T-Lymphocyte Subsets/METABOLISM  T-Lymphocytes/*METABOLISM
       Transfection/*METHODS  Tumor Cells, Cultured/*METABOLISM  JOURNAL
       ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

