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Clinical and Diagnostic Laboratory Immunology, May 2000, p. 510-514, Vol. 7, No. 3
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Apoptosis of Primary-Culture Rat Microglial Cells Induced by Pathogenic Acanthamoeba spp.

Ho-Joon Shin,1,* Myung-Soo Cho,1 Hyung-Il Kim,1 Millina Lee,1 Sun Park,1 Seonghyang Sohn,2 and Kyung-Il Im3

Department of Microbiology1 and Laboratory of Cell Biology, Institute for Medical Science,2 Ajou University School of Medicine, Suwon 442-749, and Department of Parasitology, College of Medicine, Yonsei University, Seoul 121-752,3 Korea

Received 11 August 1999/Returned for modification 26 October 1999/Accepted 18 January 2000

To determine whether trophozoites and lysates of pathogenic Acanthamoeba spp. induce apoptosis in primary-culture microglial cells, transmission electron microscopic (TEM) examinations, assessment of DNA fragmentation by agarose gel electrophoresis, and the TdT-mediated dUTP nick-end labeling assay were performed. When a trophozoite of pathogenic Acanthamoeba culbertsoni came in contact with a microglial cell, the digipodium was observed by TEM. Nuclear chromatin condensation was observed in 10% of microglial cells, while it was not revealed when they were cocultured with weakly pathogenic Acanthamoeba royreba trophozoites. DNA fragmentation in microglial cells cocultured with the A. culbertsoni lysate was detected by electrophoresis, showing DNA ladder formation, whereas it was hardly observed in microglial cells cocultured with A. royreba. DNA fragmentation of microglial cells was also confirmed by flow cytometry analysis. The fluorescence of TdT-stained apoptotic bodies became intensely visible with microglial cells cocultured with the A. culbertsoni lysate. In contrast, with microglial cells cocultured with the A. royreba lysate, only a background level of fluorescence of TdT-stained apoptotic bodies was detected. These results suggest that some rat microglial cells cocultured with pathogenic A. culbertsoni undergo cytopathic changes which show the characteristics of the apoptotic process, such as nuclear condensation and DNA fragmentation.

* Corresponding author. Mailing address: Department of Microbiology, Ajou University School of Medicine, Suwon 442-749, Korea. Phone: (82) 331-219-5076. Fax: (82) 331-219-5079. E-mail: hjshin{at}madang.ajou.ac.kr.

Clinical and Diagnostic Laboratory Immunology, May 2000, p. 510-514, Vol. 7, No. 3
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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