Nonbilayer Phospholipid Arrangements Are Toll-Like Receptor-2/6 and TLR-4 Agonists and Trigger Inflammation in a Mouse Model Resembling Human Lupus

Figure 5

C3, C5, TLR-4, and IFN-β are overexpressed in the spleens of mice with a disease resembling human lupus. The complement proteins C3 and C5 (a), the fragments C3a and C5a (b) (produced by C3 and C5 cleavage), and IFN-β (c) were detected in a cell-free extract from the spleen of mice injected with TS buffer, Control I group (CI), egg-yolk phosphatidylcholine/egg-yolk phosphatidic acid (50 nmol phosphatidic acid in 50 μL TS) liposomes alone, Control II group (CII), or bearing Mn²⁺-induced nonbilayer phospholipid arrangements (Mn-G) or chlorpromazine-induced nonbilayer phospholipid arrangements (CPZ-G). The detection ranges were C3 (4.69–300 μg/mL), C3a (0.16–10 ng/mL), C5 (0.25–16 ng/mL), C5a (0.062–4.0 ng/mL), and IFN-β (7.8–500 pg/mL). TLR-4 (d) was detected by flow cytometry on cells obtained from fresh spleens from the four treatment groups. Bars represent the mean ± SD of three independent experiments, with three mice in each of the four groups. Data were analyzed with one-way ANOVA followed by Tukey’s multiple comparisons test. , , and . Asterisks indicate statistical significance between the control groups and the Mn or CPZ groups.

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