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Interaction of L cells and Chlamydia psittaci: entry of the parasite and host responses to its development
Abstract:
The entry and development of Chlamydia psittaci in the L cell was studied by using purified, infectious parasites at high multiplicity. Entry of the parasite was accomplished by an act of phagocytosis by the host which was independent of an adsorption stage but was temperature-dependent. Kinetic studies of phagocytosis performed with (14)C-amino acid-labeled, purified parasites indicated that the rate of phagocytosis was directly proportional to the multiplicity of inoculation. Electron microscopy of cells infected at high multiplicity with purified infectious C. psittaci showed that phagocytosed chlamydiae were segregated in a host phagocytic vacuole throughout their developmental cycle which consisted of the transition of infecting elementary bodies to reticulate bodies dividing by binary fission, followed by the reemergence of a population of elementary bodies. The process of the transition was examined and a proposed sequence of intermediate bodies is presented. In isopycnic gradients of fractionated, infected L cells, chlamydial phagocytic vacuoles were apparent as a dense band distinct from lysosome and mitochondrion peaks, as indicated by acid phosphatase and cytochrome oxidase activities. Chlamydiae inactivated by heat or neutralized by antiserum were phagocytosed and appeared in lysosomes within 12 hr after infection according to electron microscopy; however, chlamydiae which were continuously inhibited in their development by chloramphenicol were retained intact in the cell for 24 hr without lysosomal response. The possibility of a lysosomal inhibitor on the native parasite is discussed.
Insights
Chlamydia psittaci enters host L cells via temperature-dependent phagocytosis, not adsorption. The bacteria develop within a unique vacuole, avoiding lysosomal degradation, suggesting a potential parasite-derived inhibitor.
Area of Science:
- Cell Biology
- Microbiology
- Infectious Diseases
Background:
- Chlamydia psittaci is an obligate intracellular bacterium.
- Understanding Chlamydia entry and development is crucial for combating infections.
Purpose of the Study:
- To investigate the entry and intracellular development of Chlamydia psittaci in L cells.
- To characterize the host-cell vacuole containing Chlamydia and its developmental stages.
Main Methods:
- Infection of L cells with purified, infectious Chlamydia psittaci at high multiplicity.
- Kinetic studies using radiolabeled parasites.
- Electron microscopy for ultrastructural analysis.
- Cell fractionation and isopycnic gradient centrifugation to identify vacuoles.
Main Results:
- Chlamydia entry is via temperature-dependent phagocytosis, independent of an adsorption stage.
- Phagocytosis rate is proportional to inoculum multiplicity.
- Infectious Chlamydia develop within a distinct phagocytic vacuole, progressing from elementary bodies to reticulate bodies and back.
- Inactivated or neutralized Chlamydia are routed to lysosomes, while developing Chlamydia are not, suggesting a lysosomal inhibitor.
Conclusions:
- Chlamydia psittaci utilizes a unique phagocytic pathway for entry and development.
- The bacteria's developmental cycle occurs within a specialized vacuole, evading lysosomal degradation.
- A potential lysosomal inhibitor produced by native Chlamydia is proposed.