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Infection and Immunity, September 1999, p. 4713-4719, Vol. 67, No. 9
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Isolation of Recombinant Protective Helicobacter pylori Antigens

D. Hocking,1,* E. Webb,1 F. Radcliff,2,dagger L. Rothel,1 S. Taylor,1 G. Pinczower,1 C. Kapouleas,1 H. Braley,1 A. Lee,2 and C. Doidge1

Research and Development Division, CSL Limited, Parkville, Victoria, Australia 3052,1 and School of Microbiology and Immunology, University of New South Wales, Sydney, New South Wales, Australia 20522

Received 15 March 1999/Returned for modification 23 April 1999/Accepted 21 June 1999

A total of seven clones producing both new and previously described Helicobacter pylori proteins were isolated from a library of H. pylori genomic DNA. The screening approach by which these proteins were detected relied on the use of antisera raised in mice vaccinated with Helicobacter felis sonicate plus cholera toxin, a regimen which protects mice from H. pylori challenge. This strategy was designed to maximize the possibility of obtaining antigens which might be capable of conferring protection from H. pylori infection. Two of the clones were shown to encode the urease enzyme and the heat shock protein HspB, which have already been identified as protective antigens. The other five clones were sequenced, protein coding regions were deduced, and these sequences were amplified by PCR for incorporation into Escherichia coli expression vectors. The proteins produced from these expression systems were purified to allow testing for protective efficacy in an H. pylori mouse model. All five proteins were able to facilitate the clearance of a challenge with H. pylori, as judged by an assay of gastric urease activity and light microscopy on stomach sections. These results clearly indicate that the screening strategy has successfully identified candidate vaccine antigens.

* Corresponding author. Mailing address: CSL Limited, 45 Poplar Rd., Parkville, Victoria, Australia 3052. Phone: 61-3-9389-1525. Fax: 61-3-9388-2063. E-mail: dianna_hocking{at}csl.com.au.

dagger Present address: Department of Haematology and Oncology, Royal Children's Hospital, Parkville, Victoria, Australia 3052.

Infection and Immunity, September 1999, p. 4713-4719, Vol. 67, No. 9
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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Copyright © 1999 by the American Society for Microbiology. All rights reserved.