Summary
After streptococcal pyrogenic exotoxin B (SPE B) induces apoptosis, its fate is unknown. Using confocal time-course microscopy at 37 °C, we detected green fluorescence 20 min after adding FITC-SPE B. Orange fluorescence, an indication of co-localization of SPE B with lysosomes which were labeled with a red fluorescent probe, was maximal at 40 min and absent by 60 min. SPE B was co-precipitated with clathrin, which is consistent with endocytotic involvement. Western blotting assay also indicated that uptake of SPE B was maximal at 40 min and disappeared after 60 min. However, in the presence of chloroquine, a lysosome inhibitor, the uptake of SPE B was not detectable. The disappearance of TCA-precipitated FITC-SPE B was parallel to the appearance of TCA soluble FITC-SPE B; in the presence of chloroquine, however, no SPE B degradation occurred. Chloroquine increased the level of SPE B-induced apoptosis by inhibiting the degradation of SPE B. These results suggest that the internalization and degradation of SPE B in cells may be a host defense system that removes toxic substances by sacrificing the exposed cells.
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This work was supported by Grants NHRI-EX 91-9027SP from the National Health Research Institute and NSC92-3112-B006-006 from the National Science Council, Taiwan.
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Chang, CW., Tsai, WH., Chuang, WJ. et al. The fate of SPE B after internalization and its implication in SPEB-induced apoptosis. J Biomed Sci 14, 419–427 (2007). https://doi.org/10.1007/s11373-007-9154-6
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DOI: https://doi.org/10.1007/s11373-007-9154-6