Characterization of the Chlamydia trachomatis vacuole and its interaction with the host endocytic pathway in HeLa cells.
Chlamydia trachomatis, an obligate intracellular parasite and a major human pathogen, invades eukaryotic host cells and replicates within a membrane-bound compartment (termed the vacuole or inclusion) in the cytoplasm of the host cell. In this report, we describe in detail the characteristics of the vacuole throughout the chlamydial life cycle in terms of the endocytic pathway, as determined by epifluorescent and confocal immunofluorescence microscopy. By indirect immunofluorescence, the transferrin receptor (TfR), a component of early endosomes, and the cation-independent mannose-6-phosphate receptor (CI-M6PR), a component of late endosomes, were found in close association with the chlamydial vacuole as early as 4 h postinfection (hpi) and as late as 20 hpi. Fluorescein isothiocyanate (FITC)-labeled Tf was also found to colocalize with the vacuole at 4, 12, and 20 hpi, indicating that exogenously added ligands can be transported to the region of the vacuole. Antibodies to several different lysosomal proteins failed to label the chlamydial vacuole at any time point during the life cycle. Indirect immunofluorescence of cells infected with chlamydiae stained with an antibody to the trans-Golgi network (TGN) protein TGN38 demonstrated that in infected cells, the integrity and structure of the TGN was altered. The rates of Tf recycling in infected and uninfected cells were compared by fluorescence microscopy and quantitated with 125I-Tf. While the rate of FITC-Tf recycling from endocytic compartments in chlamydia-infected cells did not appear different from that of uninfected cells, a small pool of FITC-Tf that had accumulated adjacent to the chlamydial vacuole recycled at a slower rate. Quantitation of Tf recycling with 125I-Tf showed that Tf was recycled more slowly in infected cells than in uninfected cells. The altered distribution of several endocytic pathway markers and the slowed Tf recycling are consistent with the hypothesis that the chlamydial vacuole interacts with the endocytic pathway of the host. These results furthermore suggest that the chlamydial vacuole does not correspond to a canonical endocytic compartment but that it is a unique and dynamic organelle that shares several characteristics with recycling endosomes of the host cell. Interactions with the early and/or late endosomal compartments, in addition to the Golgi apparatus, may provide a source of membrane or nutrients for the replicating organisms.
van Ooij
C
C
Biomedical Sciences Program, University of California, San Francisco 94143-0654, USA.
Apodaca
G
G
Engel
J
J
D015415:Biomarkers; D002690:Chlamydia Infections / Q000378:metabolism / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000166:cytology* / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism / Q000382:microbiology; D006023:Glycoproteins; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000378:metabolism / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000378:metabolism; D008565:Membrane Proteins; D011990:Receptors, Transferrin / Q000378:metabolism; D013045:Species Specificity; D014617:Vacuoles / Q000378:metabolism* / Q000382:microbiology*
D015415:Biomarkers; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011990:Receptors, Transferrin; C072237:TGOLN2 protein, human