Plasmid DNA activates murine macrophages to induce inflammatory cytokines in a CpG motif-independent manner by complex formation with cationic liposomes

Yasuda, Kei; Ogawa, Yoshiyuki; Kishimoto, M.; Takagi, Toshihide; Hashida, Mitsuru; Takakura, Yoshinobu

doi:10.1016/s0006-291x(02)00210-3

Cited by 66 publications

(69 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We calculated the amounts of the methylated and unmethylated intact plasmids, and the methylated and unmethylated DNA fragments, at various time points (Table 1). The methylated DNA fragments were 35-, 19-, 9-, and 7-fold more abundant than the methylated full-length plasmid at 4,8,24, and 48 h, respectively, after the administration. Taken together, we concluded that the degraded DNA fraction contained more methylation in the promoter region.…”

Section: Methylation Status Of Intranuclear Dnamentioning

confidence: 94%

“…This method enables the delivery of exogenous DNA into the nuclei of the liver, without the aid of cationic lipids and polymers, which could potentially affect the quantitation of the intranuclear exogenous DNA. The livers were harvested after 4,8,24,48,72,96, and 168 h, and the luciferase activity was measured. The amounts of exogenous DNA at the same time points were examined by quantitative polymerase chain reaction (Q-PCR) after isolation of the nuclei.…”

Section: Time Courses Of the Luciferase Activity And The Amount Ofmentioning

confidence: 99%

See 1 more Smart Citation

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

2006

View full text Add to dashboard Cite

The intranuclear disposition of exogenous DNA is highly important for the therapeutic effects of the administrated DNA. Naked luciferase-plasmid DNA was delivered into mouse liver by a hydrodynamics-based injection, and the amounts of intranuclear plasmid DNA, luciferase, and its mRNA were quantitated at various time points. Methylation of the promoter of the luciferase gene was also analyzed. Expression efficiency from one copy of the exogenous DNA dramatically decreased over time, and the DNA was methylated and degraded into fragments. Unexpectedly, methylation of the intact plasmid DNA was low and did not increase over time. Rather, the fragmented DNA was methylated more frequently than the intact plasmid. These results suggest that the CpG methylation and the degradation of exogenous DNA, and its 'silencing', occurred in parallel in the nucleus.

show abstract

Section: Methylation Status Of Intranuclear Dnamentioning

confidence: 94%

Section: Time Courses Of the Luciferase Activity And The Amount Ofmentioning

confidence: 99%

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

2006

View full text Add to dashboard Cite

show abstract

“…DNA devoid of CpG motifs does not induce proinflammatory cytokine synthesis by macrophages in vitro. 10 In contrast, CpG motifs have immunostimulatory effects including activation of the NFkB pathway, enhanced synthesis by monocytes of cytokines such as TNF-a or IL-6, and activation of NK cells. 11 Unmethylated CpG motifs also open up the adaptive immunity pathway by inducing the synthesis of IFNg, IL-12, and IL-18 and by activating the Th1 pathway.…”

Section: Introductionmentioning

confidence: 99%

Immune responses to gene therapy vectors: influence on vector function and effector mechanisms

2004

View full text Add to dashboard Cite

“…Yasuda et al showed that lipoplexes activated murine macrophages to induce the cytokines in a CpG motif-independent manner. 38 McLachlan et al demonstrated that lipoplexes containing methylated plasmid DNA, as well as lipoplexes containing unmethylated plasmid DNA, caused severe inflammation in the lung, suggesting that other factors involved in the inflammation are unaffected by methylation of plasmid DNA. 39 Other methylated motifs different from the conventional CpG motif (5 0 -Pur-Pur-CG-Pyr-Pyr-3 0 ) may also contribute to lipoplex-mediated inflammatory response.…”

Section: Discussionmentioning

confidence: 99%

Therapeutic effect of intravenous delivery of lipoplexes containing the interferon-β gene and poly I: poly C in a murine lung metastasis model

et al. 2003

Self Cite

View full text Add to dashboard Cite

We have evaluated and compared the efficacy of systemic administration of lipoplex formulations containing plasmids encoding IFN-b or IFN-g, and a synthetic double-strand RNA poly I:poly C (pI:pC), a type I IFN inducer, in a lung metastasis model in which colon carcinoma CT-26 cells were inoculated intravenously into immunocompatible mice. Injection of lipoplexes containing plasmid DNA, regardless of IFN gene insertion, stimulated a transient increase in the serum concentration of proinflammatory cytokines such as tumor necrosis factor (TNF)-a and IFN-g, while injection of lipoplexes containing pI:pC led to a low level of TNF-a and undetectable IFN-g production. Furthermore, injection of these lipoplexes containing plasmids resulted in the production of a mixture of type I and type II IFNs, partly derived from the inserted IFN genes, in lung tissue cultures. In tumor-prophylactic experiments, intravenous injection of lipoplexes containing plasmid, regardless of IFN gene insertion, showed a significant reduction in lung metastatic nodules probably due to proinflammatory cytokines such as TNF-a and IFN-g nonspecifically induced by the CpG motifs in the plasmid and the type I IFNs produced. On the other hand, the antimetastatic effect of pI:pC-lipoplex seemed to be due mainly to IFN-b induced by pI:pC. In established lung metastasis experiments, a single intravenous administration of lipoplexes containing IFN-b gene or pI:pC, but not other lipoplexes, showed a significant therapeutic effect on the tumor metastasis: reduction in tumor nodules and prolongation of survival time of tumor-burden mice. The therapeutic effects were specifically impaired by anti-IFN-b antibody treatment, indicating that IFN-b produced by the lipoplexes played an important role in the suppression of established metastatic lung tumors. Thus, the local IFN-b in the lung delivered by intravenous administration of lipoplex containing IFN-b gene or pI:pC may be a convenient and useful method of inhibiting established metastatic lung tumors.

show abstract

Plasmid DNA activates murine macrophages to induce inflammatory cytokines in a CpG motif-independent manner by complex formation with cationic liposomes

Cited by 66 publications

References 27 publications

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

Immune responses to gene therapy vectors: influence on vector function and effector mechanisms

Therapeutic effect of intravenous delivery of lipoplexes containing the interferon-β gene and poly I: poly C in a murine lung metastasis model

Contact Info

Product

Resources

About