Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo splenic DC-specific transfection of mRNA/H 18 NPs and in vitro protein corona analysis of mRNA/H 18 NPs. (A) EGFP protein expression in splenic cell subsets of C57BL/6J mice 24 h post intravenous injection of different formulations. (B) The top 5 most abundant plasma proteins adsorbed on mRNA/H 18 NPs (C3: Complement C3; Ighm: Immunoglobulin heavy constant mu; Hbat1: Alpha-globin; Itih4: Inter alpha-trypsin inhibitor, heavy chain 4; Cnn2: Calponin). (C) Heatmap plot of major proteins in the protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. PBS group was used as a negative control. (D) Quantification of major adsorbed protein categories of different formulations. (E) Complement C3 abundance in protein corona adsorbed on mRNA/MC3-LNPs and mRNA/H 18 NPs. (F) Bioluminescence images of major organs and (G) Quantification of total bioluminescence flux in the spleen from C57BL/6J mice 6 h after intravenous injection of mLuc/H 18 NPs (mLuc dose of 0.25 mg kg −1 ). Mice were pre-treated with cobra venom factor (CVF) or PBS. (H) Fluorescence quantification of Cy5 mRNA delivered by uncoated or complement C3-coated Cy5-mRNA/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. (I) Bioluminescence intensity of luciferase protein translated from mRNA delivered by uncoated or complement C3-coated mLuc/H 18 NPs in BMDCs. BMDCs were pre-incubated with anti-CD11b (CR3) or anti-IgG blocking antibody. Data were shown as mean ± SD (n = 3).

Article Snippet: Cobra venom factor (CVF) was purchased from TargetMol (MA, USA).

Techniques: In Vivo, Transfection, In Vitro, Expressing, Injection, Clinical Proteomics, Negative Control, Combined Bisulfite Restriction Analysis Assay, Fluorescence, Incubation, Blocking Assay, Luciferase

Journal: Journal of Neuroinflammation

Article Title: Complement component C3a plays a critical role in endothelial activation and leukocyte recruitment into the brain

doi: 10.1186/s12974-016-0485-y

Figure Lengend Snippet: Effects of complement depletion on leukocyte recruitment into the brain parenchyma after i.c.v. LPS injection. a WT C57BL/6J mice were treated i.c.v. with LPS, and C3 levels in the brain (without dilution, cleared from blood proteins) and plasma (diluted in 1:100) were determined by Western blotting. b WT C57BL/6J mice received i.p. injections of CVF (10 μg/mouse) or PBS. After 4 h, WT C57BL/6J mice ( n = 4) received i.c.v. LPS injection, and after 4 h, plasma and brain samples were then collected for C3 Western blotting analysis. At 4 h after CVF injection, WT C57BL/6J mice ( n = 4) received i.c.v. LPS injection, and plasma samples were collected for C3a ELISA assay ( c ). At 4 h after i.p. injection of CVF to deplete complement from the blood circulation, mice received i.c.v. LPS injection. Then, after 4 h, rolling and adhesion leukocytes were quantified using intravital microscopy ( d ). e P-selectin and E-selectin mRNA expressions were measured using real-time PCR. f VCAM-1 and E-selectin protein levels in the brain were examined by Western blotting ( n = 4). Images are representative of four experiments. The ratio of VCAM-1 and E-selectin to β-actin was determined through densitometry. The results are shown as the means ± SEM. * P < 0.05, ** P < 0.01; n ≥ 4 for all groups

Article Snippet: LPS injection, hypocomplementemia was induced through the i.p. injection of cobra venom factor (CVF) (Biogen Sci & Tech Co., Kunming, China) in PBS into C57BL/6J mice at a dose of 10 μg/mouse (15 U/mouse).

Techniques: Injection, Clinical Proteomics, Western Blot, Enzyme-linked Immunosorbent Assay, Intravital Microscopy, Real-time Polymerase Chain Reaction