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Infection and Immunity, January 2000, p. 30-37, Vol. 68, No. 1
0019-9567/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Increased Expression of Periplasmic Cu,Zn Superoxide Dismutase Enhances Survival of Escherichia coli Invasive Strains within Nonphagocytic Cells

Andrea Battistoni,1,* Francesca Pacello,1 Silvia Folcarelli,1 Maria Ajello,2 Giovanna Donnarumma,2 Rita Greco,2 Maria Grazia Ammendolia,3 Danièle Touati,4 Giuseppe Rotilio,1 and Piera Valenti2

Department of Biology, Università di Roma "Tor Vergata," 00133 Rome,1 Istituto Superiore di Sanità, 00161 Rome,3 and Istituto di Microbiologia, II Università di Napoli, 80138 Naples,2 Italy, and Institut Jacques Monod, CNRS, Universités Paris 6 and 7, Paris, France4

Received 2 August 1999/Returned for modification 10 September 1999/Accepted 21 October 1999

We have studied the influence of periplasmic Cu,Zn superoxide dismutase on the intracellular survival of Escherichia coli strains able to invade epithelial cells by the expression of the inv gene from Yersinia pseudotuberculosis but unable to multiply intracellularly. Intracellular viability assays, confirmed by electron microscopy observations, showed that invasive strains of E. coli engineered to increase Cu,Zn superoxide dismutase production are much more resistant to intracellular killing than strains containing only the chromosomal sodC copy. However, we have found only a slight difference in survival within HeLa cells between a sodC-null mutant and its isogenic wild-type strain. Such a small difference in survival correlates with the very low expression of this enzyme in the wild-type strain. We have also observed that acid- and oxidative stress-sensitive E. coli HB101(pRI203) is more rapidly killed in epithelial cells than E. coli GC4468(pRI203). The high mortality of E. coli HB101(pRI203), independent of the acidification of the endosome, is abolished by the overexpression of sodC. Our data suggest that oxyradicals are involved in the mechanisms of bacterial killing within epithelial cells and that high-level production of periplasmic Cu,Zn superoxide dismutase provides bacteria with an effective protection against oxidative damage. We propose that Cu,Zn superoxide dismutase could offer an important selective advantage in survival within host cells to bacteria expressing high levels of this enzyme.

* Corresponding author. Mailing address: Department of Biology, Università di Roma "Tor Vergata," Via della Ricerca Scientifica, 00133 Rome, Italy. Phone: 39-0672594372. Fax: 39-0672594311. E-mail: andrea.battistoni{at}uniroma2.it.

Infection and Immunity, January 2000, p. 30-37, Vol. 68, No. 1
0019-9567/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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