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Antimicrobial Agents and Chemotherapy, May 1998, p. 1015-1021, Vol. 42, No. 5
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Biological Characterization of Endotoxins Released from Antibiotic-Treated Pseudomonas aeruginosa and Escherichia coli

Teruo Kirikae,1,* Fumiko Kirikae,1 Shinji Saito,1 Kaoru Tominaga,1 Hirohi Tamura,2 Yayoi Uemura,2 Takashi Yokochi,3 and Masayasu Nakano1

Department of Microbiology, Jichi Medical School, Minamikawachi-machi, Tochigi-ken 329-0498,1 Tokyo Research Institute, Seikagaku Cooperation, Higashiyamato, Tokyo 207-0021,2 and Department of Microbiology and Immunology, Aichi Medical University, Aichi 480-1195,3 Japan

Received 28 July 1997/Returned for modification 15 December 1997/Accepted 15 February 1998

The supernatants taken from Pseudomonas aeruginosa and Escherichia coli cultures in human sera or chemically defined M9 medium in the presence of ceftazidime (CAZ) contained high levels of endotoxin, while those taken from the same cultures in the presence of imipenem (IPM) yielded a very low level of endotoxin. The biological activities of endotoxin in the supernatants were compared with those of phenol water-extracted lipopolysaccharide (LPS). The endotoxin released from the organisms as a result of CAZ treatment (CAZ-released endotoxin) contained a large amount of protein. The protein, however, lacked endotoxic activity, since the endotoxin did not show any in vivo toxic effects in LPS-hyporesponsive C3H/HeJ mice sensitized with D-(+)-galactosamine (GalN) or any activation of C3H/HeJ mouse macrophages in vitro. The activities of CAZ- and IPM-released endotoxin (as assessed by a chromogenic Limulus test) were fundamentally the same as those of P. aeruginosa LPS, since their regression lines were parallel. The CAZ-released endotoxin was similar to purified LPS with respect to the following biological activities in LPS-responsive C3H/HeN mice and LPS-hyporesponsive C3H/HeJ mice: lethal toxicity in GalN-sensitized mice, in vitro induction of tumor necrosis factor- and NO production by macrophages, and mitogen-activated protein kinase activation in macrophages. The macrophage activation by CAZ-released endotoxin as well as LPS was mainly dependent on the presence of serum factor and CD14 antigen. Polymyxin B blocked the activity. These findings indicate that the endotoxic activity of CAZ-released endotoxin is due primarily to LPS (lipid A).

* Corresponding author. Mailing address: Department of Microbiology, Jichi Medical School, Minamikawachi-machi, Tochigi-ken 329-0498, Japan. Phone: 81-285-44-2111, ext. 3160. Fax: 81-285-44-1175. E-mail: tkirikae{at}jichi.ac.jp.

Antimicrobial Agents and Chemotherapy, May 1998, p. 1015-1021, Vol. 42, No. 5
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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