Mannan-Binding Lectin Inhibits Candida albicans-Induced Cellular Responses in PMA-Activated THP-1 Cells through Toll-Like Receptor 2 and Toll-Like Receptor 4
Figure 5
MBL decreases C. albicans-stimulated NF-κB binding activity and translocation in PMA-activated THP-1 cells.
(A) MBL inhibits C. albicans-stimulated NF-κB DNA-binding activity. PMA-activated THP-1 cells (5×105 cells/sample) were stimulated with heat-inactivated Y or H cells of C. albicans at a Candida/THP-1 ratio of 2:1 in the presence of the indicated concentrations of MBL, anti-MBL pAb and MBL, or HSA for 1 h, then harvested to prepare nuclear extracts. The nuclear extracts were mixed with radiolabeled NF-κB oligonucleotide probe and analyzed with EMSA. (B) NF-κB translocation is inhibited by MBL in PMA-activated THP-1 cell. PMA-activated THP-1 cells (5×105 cells/sample) were stimulated with HK Y or H cells of C. albicans at a Candida/THP-1 ratio of 2:1 in the presence of 20 µg/ml MBL, anti-MBL pAb and MBL, or HSA for 1 h, then the cells were harvested to prepare nuclear extracts. The proteins in the nuclei-free supernatants were separated by 10% SDS-PAGE, followed by transfer to a nitrocellulose membrane. After blocking, the membrane was incubated with the NF-κB-specific mouse anti-human mAb p65, followed by HRP-conjugated secondary antibody. ECL was used to visualize the protein bands. β-actin was used as an internal control.