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Infection and Immunity, June 2001, p. 4134-4140, Vol. 69, No. 6
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.6.4134-4140.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

By Releasing ADP, Acanthamoeba castellanii Causes an Increase in the Cytosolic Free Calcium Concentration and Apoptosis in Wish Cells

A. Mattana,1,* M. G. Tozzi,1 M. Costa,1 G. Delogu,2 P. L. Fiori,2 and P. Cappuccinelli2

Department of Pharmacological Sciences1 and Department of Biomedical Sciences, Division of Experimental and Clinical Microbiology,2 University of Sassari, Sassari, Italy

Received 20 November 2000/Returned for modification 29 December 2000/Accepted 28 February 2001

The role played by soluble molecules that may participate in acanthamoebal cytopathogenicity has yet to be fully characterized. We demonstrate here that Acanthamoeba castellanii trophozoites constitutively release ADP in the medium. Cell-free supernatants prepared from A. castellanii, by interaction with specific P2y2 purinoceptors expressed on the Wish cell membrane, caused a biphasic rise in [Ca2+]i, extensive cell membrane blebbing, cytoskeletal disorganization, and the breakdown of nuclei. Cell damage induced by amoebic supernatants was blocked by the P2y2 inhibitor Suramin. The same results were found in Wish cells exposed to purified ADP. These findings suggest that pathogenic free-living A. castellanii may have a cytopathic effect on human epithelial cells through ADP release, by a process that begins with a rise of cytosolic free-calcium concentration, and culminates in apoptosis.

* Corresponding author. Mailing address: Dipartimento di Scienze del Farmaco, Via Muroni 23/A, 07100 Sassari, Italy. Phone: 0039-79-228744/228711. Fax: 0039-79-228712. E-mail: dsfanto{at}ssmain.uniss.it.

Infection and Immunity, June 2001, p. 4134-4140, Vol. 69, No. 6
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.6.4134-4140.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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