Effect of Cationic Liposomes on Intracellular Trafficking and Efficacy of Antisense Oligonucleotides in Mouse Peritoneal Macrophages

Takagi, Toshihide; Hashiguchi, Miwa; Hiramatsu, Tetsuhiko; Yamashita, Fumiyoshi; Takakura, Yoshinobu; Hashida, Mitsuru

doi:10.3109/10611869909085519

Cited by 7 publications

(2 citation statements)

References 25 publications

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“…Indeed, these formulations tended to increase TNF-a secretion by microglia and macrophages or, in the case of polyethylenamine, were cytotoxic at longer transfection times. The use of several cationic lipids to deliver oligonucleotides to macrophages, including some used in the current study, has been shown previously to result in encapsulated oligonucleotide accumulation in lysosomal compartments (Takagi et al, 2000). In contrast, TNF-a ASOs alone, but not a control mixed base oligonucleotide, potently inhibited TNF-a production, resulting in a dramatic morphological change that involved a diminution of cell body size from a large amoeboid shape and the formation of extensions that produced either spindle-or stellate-shaped cells.…”

Section: Discussionmentioning

confidence: 81%

“…Indeed, these formulations tended to increase TNF‐α secretion by microglia and macrophages or, in the case of polyethylenamine, were cytotoxic at longer transfection times. The use of several cationic lipids to deliver oligonucleotides to macrophages, including some used in the current study, has been shown previously to result in encapsulated oligonucleotide accumulation in lysosomal compartments (Takagi et al. , 2000).…”

Section: Discussionmentioning

confidence: 85%

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Inhibition of tumour necrosis factor‐α by antisense targeting produces immunophenotypical and morphological changes in injury‐activated microglia and macrophages

Pearse

Pereira

Stolyarova

et al. 2004

Eur J of Neuroscience

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Microglia respond in a stereotypical pattern to a diverse array of pathological states. These changes are coupled to morphological and immunophenotypical alterations and the release of a variety of reactive species, trophic factors and cytokines that modify both microglia and their cellular environment. We examined whether a microglial-produced cytokine, tumour necrosis factor-alpha (TNF-alpha), was involved in the maintenance of microglial activation after spinal cord injury by selective inhibition using TNF-alpha antisense deoxyoligonucleotides (ASOs). Microglia and macrophages harvested from 3 d post-contused rat spinal cord were large and rounded (86.3 +/- 9.6%). They were GSA-IB4-positive (GSA-IB4(+)) (Griffonia simplicifolia lectin, microglia specific; 94.8 +/- 5.1%), strongly OX-42 positive (raised against a type 3 complement/integrin receptor, CD11b; 78.9 +/- 9.1%), ED-1 positive (a lysosomal marker shown to correlate well with immune cell activation; 97.2 +/- 2.6%) and IIA positive (antibody recognizes major histocompatibility complex II; 57.2 +/- 5.6%), indicative of fully activated cells, for up to 48 h after plating. These cells also secreted significant amounts of TNF-alpha (up to 436 pg/microg total protein, 16 h). Fluoroscein isothiocyanate-labelled TNF-alpha ASOs (5, 50 and 200 nm) added to the culture medium were taken up very efficiently into the cells (> 90% cells) and significantly reduced TNF-alpha production by up to 92% (26.5 pg/microg total protein, 16 h, 200 nm TNF-alpha ASOs). Furthermore, few of the treated cells at this time were round (5.4 +/- 2.7%), having become predominantly spindle shaped (74.9 +/- 6.3%) or stellate (21.4 +/- 2.7%); immunophenotypically, although all of them remained GSA-IB4 positive (91.6 +/- 6.2%), many were weakly OX-42 positive and few expressed either ED-1 (12.9 +/- 2.5%) or IIA (19.8 +/- 7.4%). Thus, the secretion of TNF-alpha early in spinal cord injury may be involved in autoactivating microglia/macrophages. However, at the peak of microglial activation after injury, the activation state of microglia/macrophages is not stable and this process may still be reversible by blocking TNF-alpha.

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Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 85%