FIGURE 9.

Effects of disruption of overall shapes of ICWs on immunomodulatory abilities of ICWs. (A) Scanning electron microscopy of ICWs and DCWs of L. plantarum TUA 5099L. (B) Uptake of ICWs or DCWs. J774.1 cells were incubated with fluorescein-labeled ICWs or disrupted ICWs (DCWs) of L. plantarum TUA 5099 for 30 min and fluorescent signals in the cells were analyzed. Gray and open histograms indicate untreated and fluorescein-labeled cell walls treated cells, respectively. Representative images under florescent microscopy were also shown. (C) Reduction of uptake of DCWs by HF treatment. J774.1 cells were incubated with fluorescein-labeled DCWs (solid line) or HF-treated DCWs (dashed line) for 30 min and fluorescent signals in the cells were analyzed. Gray histograms indicate cells without DCW treatment. (D) Elicitation of IL-12 secretion from macrophages. J774.1 cells were cultured with ICWs (10 μg/ml) or DCWs (100 μg/ml) for 24 h and IL-12 secreted in culture supernatants was determined. Each bar is the mean ± SE for three independent experiments. (E) Effect of cytochalasin D on uptake of DCWs. J774.1 cells were treated with cytochalasin D in complete RPMI1640 containing 0.1% DMSO for 30 min at 37°C. Fluorescein-labeled DCWs were added and the mixture was incubated for 30 min. Fluorescent signals in the cells were analyzed. Solid and dashed lines indicate untreated and cytochalasin D-treated cells, respectively. Gray histograms indicate cells without DCW treatment.