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. 2009 Dec 8;18(2):370–376. doi: 10.1038/mt.2009.271

Figure 3.

Figure 3

Systemic delivery of siCCR5 with LFA-1 I-tsNPs silences CCR5 expression in vivo. (a) siCCR5 uptake was monitored using RT-PCR in peripheral blood T cells (CD3+), B cells (CD19+), and monocytes (CD14+) in humanized mice. n.d., not detected. (b) The CCR5 levels in peripheral blood CD14+ monocytes were measured by RT-PCR before, and 3 and 10 days after siRNA treatment from blood collected. Reduction in CCR5 expression was calculated as a percentage of initial expression level before siRNA injection. siCCR5 (open bar) and siLuc (solid bar). (c) Expression of IFN responsive genes relative to β-actin were analyzed by quantitative RT-PCR in PBMC treated with siLuc delivered as indicated. Poly (I:C) was used as a positive control to induce IFN responses. (d) Freshly isolated PBMCs were treated with LFA-1 I-tsNP-encapsulated siLuc, siCCR5, or siβgal, and TNF-α release was determined after 24 hours of stimulation. Lipofectamine 2000–encapsulated siβgal was used as a positive control. Empty liposomes and naked siRNA did not induce detectable cytokines. Values are mean ± SD of triplicate cultures. IFN, interferon; LFA-1, lymphocyte function–associated antigen-1; TNF-α, tumor necrosis factor-α.