The fate of SPE B after internalization and its implication in SPEB-induced apoptosis

J Biomed Sci. 2007 May;14(3):419-27. doi: 10.1007/s11373-007-9154-6. Epub 2007 Mar 23.

Abstract

After streptococcal pyrogenic exotoxin B (SPE B) induces apoptosis, its fate is unknown. Using confocal time-course microscopy at 37 degrees 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis / immunology
  • Bacterial Proteins / metabolism
  • Bacterial Proteins / toxicity*
  • Cells, Cultured
  • Exotoxins / metabolism
  • Exotoxins / toxicity*
  • Humans
  • Streptococcus pyogenes / enzymology
  • Streptococcus pyogenes / immunology

Substances

  • Bacterial Proteins
  • Exotoxins
  • erythrogenic toxin