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Infection and Immunity, October 2002, p. 5381-5389, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5381-5389.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Contribution of Membrane-Damaging Toxins to Bacillus Endophthalmitis Pathogenesis

Michelle C. Callegan,1,2,3* Daniel C. Cochran,1,3 Scott T. Kane,1,3 Michael S. Gilmore,1,2,3 Myriam Gominet,4 and Didier Lereclus4

Department of Ophthalmology,1 Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center,2 Molecular Pathogenesis of Eye Infections Research Center, Dean A. McGee Eye Institute, Oklahoma City, Oklahoma,3 Unité de Biochimie Microbienne, Institut Pasteur, Paris cedex, France4

Received 8 April 2002/ Returned for modification 22 May 2002/ Accepted 2 June 2002

Membrane-damaging toxins are thought to be responsible for the explosive clinical course of Bacillus endophthalmitis. This study analyzed the contribution of phosphatidylinositol-specific phospholipase C (PI-PLC) and phosphatidylcholine-specific phospholipase C (PC-PLC) to the pathogenesis of experimental Bacillus endophthalmitis. Isogenic mutants were constructed by insertion of lacZ into Bacillus thuringiensis genes encoding PI-PLC (plcA) and PC-PLC (plcB). Rabbit eyes were injected intravitreally with 2 log10 CFU of strain BT407 (wild type), the PI-PLC mutant (BTplcA::lacZ), or the PC-PLC mutant (BTplcB::lacZ). The rates of decrease in retinal responses of eyes infected with the isogenic mutants were similar to that of wild type, with all infections resulting in elimination of retinal function by 18 h. Strain BT407 caused a significant increase in the latency of retinal responses at 6 h, but strains BTplcA::lacZ and BTplcB::lacZ did not. All strains elicited significant inflammatory cell influx into the anterior chamber by 12 h. Histologically, eyes infected with each strain were indistinguishable throughout the infection course. In this model, neither PI-PLC nor PC-PLC had an effect on the course or severity of experimental Bacillus endophthalmitis. Alterations in retinal responses early in infection may mark the beginnings of specific photoreceptor or glial cell dysfunction.

* Corresponding author. Mailing address: Department of Ophthalmology DMEI 418, 608 Stanton L. Young Blvd., Oklahoma City, OK 73104. Phone: (405) 271-1084. Fax: (405) 271-8781. E-mail: michelle-callegan{at}ouhsc.edu.

Editor: E. I. Tuomanen

Infection and Immunity, October 2002, p. 5381-5389, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5381-5389.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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