Journal: Journal of Controlled Release

Article Title: Alpha-2-macroglobulin loaded microcapsules enhance human leukocyte functions and innate immune response

doi: 10.1016/j.jconrel.2015.09.021

Figure Lengend Snippet: α2MG-MCs promote monocyte-derived macrophage (MDM) uptake of Zymosan and E. coli particles. Expression of LRP1 on human monocytes and MDM as quantified by flow cytometry in intact and permeabilized cells, along with CD14 for monocytes (0.5 μg/ml) and CD68 (0.5 μg/ml) for MDM. (A) Representative histograms showing augmented levels of LRP1 expression on MDM; bar graphs show cumulative data (mean ± SEM, n = 4; One way ANOVA, Bonferroni post-test, **p < 0.01 vs. MDM surface expression). (B) MDM uptake of fluorescent Zymosan (125 μg/ml, 20 min) or (C) E. coli (1 mg/ml, 1 h) particles upon MC application. Soluble α2MG (94 ng) was used for comparative purposes. Data are mean ± SEM of 4 different human donors. One way ANOVA, Bonferroni post-test, **p < 0.01 vs. vehicle, + p < 0.05 vs. 1 × 10 5 α2MG-MCs. (D) Representative confocal Z-stack images of MDM incubated with E. coli alone (vehicle) (left panel), α2MG-MCs and E. coli (central panel) or empty MCs and E. coli (right panel) as analyzed after phagocytosis. (E) Representative light microscopy images of MDM after 24 h incubation with MCs (1 × 10 5 cells with 1 × 10 5 MCs).

Article Snippet: Monocytes and MDM were assessed for both surface and intracellular expression of α2MG receptor (LRP1 or CD91, 5 μg/ml, clone A2Mr alpha-2, AbDSerotec) along with the lineage specific lineage marker: CD14 (0.5 μg/ml, clone 61D3, eBioscence) for monocytes and CD68 (0.5 μg/ml, clone Y1/82A, eBioscence) for MDM.

Techniques: Derivative Assay, Expressing, Flow Cytometry, Incubation, Light Microscopy