Interaction of cationic liposomes and their DNA complexes with monocytic leukemia cells

被引:108
作者
Pires, P
Simoes, S
Nir, S
Gaspar, R
Düzgünes, N
de Lima, MCP
机构
[1] Univ Coimbra, Fac Sci & Technol, Dept Biochem, P-3000 Coimbra, Portugal
[2] Univ Coimbra, Ctr Neurosci, P-3000 Coimbra, Portugal
[3] Univ Coimbra, Fac Pharm, Pharmaceut Technol Lab, P-3000 Coimbra, Portugal
[4] Hebrew Univ Jerusalem, Fac Agr, Dept Soil & Water Sci, IL-76100 Rehovot, Israel
[5] Univ Pacific, Sch Dent, Dept Microbiol, San Francisco, CA 94115 USA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 1999年 / 1418卷 / 01期
关键词
cationic liposome; lipoplex; membrane fusion; transfection; THP-1; cell;
D O I
10.1016/S0005-2736(99)00023-1
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cationic liposomes complexed with DNA have been used extensively as non-viral vectors for the intracellular delivery of reporter or therapeutic genes in culture and in vivo. We examined the relationship between the characteristics of the lipoplexes, their mode of interaction with monocytic THP-1 cells and their ability to transfect these cells. We determined the size and zeta potential of cationic liposomes (composed of 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP) and its mixtures with neutral lipids), and lipoplexes at different (+/-) charge ratios. As the (+/-) charge ratio of the lipoplexes decreased to (1/1), a significant reduction in zeta potential and an increase in size was observed. The increase in size resulted from fusion between liposomes promoted by DNA, as demonstrated by a lipid mixing assay, and from aggregation of the complexes. Interaction of liposomes and lipoplexes with THP-1 cells was assessed by monitoring lipid mixing ('fusion') as well as binding and cell association. While no lipid mixing was observed with the 1/2 (+/-) lipid/DNA complexes, lipoplexes with higher (+/-) charge ratios underwent significant fusion in conjunction with extensive cell binding. Liposome binding to cells was dependent on the positive charge of the liposomes, and their fusion could be modulated by the co-lipid. DOTAP/phosphatidylethanolamine (1:1) liposomes fused with THP-1 cells, unlike DOTAP/phosphatidylcholine (1:1) liposomes, although both liposome types bound to the cells to a similar extent. The use of inhibitors of endocytosis indicated that fusion of the cationic liposomes with cells occurred mainly at the plasma membrane level. The presence of serum increased the size of the cationic liposomes, but not that of the lipoplexes. Low concentrations of serum (3%) completely inhibited the fusion of cationic liposomes with cells, while inhibiting binding by only 20%. Our results suggest that binding of cationic liposomes and lipoplexes to cells is governed primarily by electrostatic interactions, whereas their fusion is regulated by the lipid composition and sterically favorable interactions with cell surface molecules. In addition our results indicate no correlation between fusion of the lipoplexes with the plasma membrane and the levels of transfection, (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:71 / 84
页数:14
相关论文
共 57 条
[1]  
BARTLETT GR, 1959, J BIOL CHEM, V234, P466
[2]  
BENNETT CF, 1992, MOL PHARMACOL, V41, P1023
[3]  
BERGMEYER H.U., 1974, METHODS ENZYMATIC AN, P574, DOI DOI 10.1016/B978-0-12-091302-2.50010-4
[4]   STABILITY AND FUSION OF LIPID VESICLES CONTAINING HEADGROUP-MODIFIED ANALOGS OF PHOSPHATIDYLETHANOLAMINE [J].
BROWN, PM ;
SILVIUS, JR .
BIOCHIMICA ET BIOPHYSICA ACTA, 1989, 980 (02) :181-190
[5]   LIPOSOME-MEDIATED CFTR GENE-TRANSFER TO THE NASAL EPITHELIUM OF PATIENTS WITH CYSTIC-FIBROSIS [J].
CAPLEN, NJ ;
ALTON, EWFW ;
MIDDLETON, PG ;
DORIN, JR ;
STEVENSON, BJ ;
GAO, X ;
DURHAM, SR ;
JEFFERY, PK ;
HODSON, ME ;
COUTELLE, C ;
HUANG, L ;
PORTEOUS, DJ ;
WILLIAMSON, R ;
GEDDES, DM .
NATURE MEDICINE, 1995, 1 (01) :39-46
[6]  
Ciccarone V., 1993, FOCUS, V15, P80
[7]   TRANSFER OF GENES TO HUMANS - EARLY LESSONS AND OBSTACLES TO SUCCESS [J].
CRYSTAL, RG .
SCIENCE, 1995, 270 (5235) :404-410
[8]   INHIBITION OF PROTEIN-KINASE C-ALPHA EXPRESSION IN MICE AFTER SYSTEMIC ADMINISTRATION OF PHOSPHOROTHIOATE ANTISENSE OLIGODEOXYNUCLEOTIDES [J].
DEAN, NM ;
MCKAY, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (24) :11762-11766
[9]   STUDIES ON THE MECHANISM OF MEMBRANE-FUSION - ROLE OF HEADGROUP COMPOSITION IN CALCIUM-INDUCED AND MAGNESIUM-INDUCED FUSION OF MIXED PHOSPHOLIPID-VESICLES [J].
DUZGUNES, N ;
WILSCHUT, J ;
FRALEY, R ;
PAPAHADJOPOULOS, D .
BIOCHIMICA ET BIOPHYSICA ACTA, 1981, 642 (01) :182-195
[10]   FUSION OF LIPOSOMES CONTAINING A NOVEL CATIONIC LIPID, N-[2,3-(DIOLEYLOXY)PROPYL]-N,N,N-TRIMETHYLAMMONIUM - INDUCTION BY MULTIVALENT ANIONS AND ASYMMETRIC FUSION WITH ACIDIC PHOSPHOLIPID-VESICLES [J].
DUZGUNES, N ;
GOLDSTEIN, JA ;
FRIEND, DS ;
FELGNER, PL .
BIOCHEMISTRY, 1989, 28 (23) :9179-9184