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

Figure 3

Antibodies found in the sera of mice with the disease resembling human lupus. Mice (10 per group) were injected with egg-yolk phosphatidylcholine/egg-yolk phosphatidic acid (2 : 1) liposomes (50 nmol phosphatidic acid in 50 μL TS buffer), alone (negative control) or incubated 30 min at 37°C with 5 mM MnCl₂ or 3 mM chlorpromazine (CPZ) to induce nonbilayer phospholipid arrangements. Mice injected with TS buffer were also used as negative control. (a) Anti-nonbilayer phospholipid arrangements, (b) anti-cardiolipin (CL), (c) anti-histone, and (d) lupus anti-coagulant antibodies were measured before and 1–4 months after liposome injection. Histograms represent the mean ± SD of three independent experiments. Control mice injected with TS buffer or with liposomes without nonbilayer phospholipid arrangements did not have detectable levels of any of these four antibodies, 1 or 4 months after the administration of TS or liposomes. ND: not detected. Data were analyzed with Kruskal-Wallis test with Dunn’s post-test (GraphPad Prism). ; . Asterisks indicate statistical significance between the antibody titers induced by liposomes incubated with CPZ and those incubated with Mn.

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