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Infection and Immunity, June 2000, p. 3448-3454, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Immune Response against a Cross-Reactive Epitope on the Heat Shock Protein 60 Homologue of Helicobacter pylori

Hiroyuki Yamaguchi,1,* Takako Osaki,2 Masanori Kai,3 Haruhiko Taguchi,1 and Shigeru Kamiya1

Department of Microbiology1 and Division of Flowcytometry,2 Kyorin University School of Medicine, Mitaka, and Leprosy Research Center, National Institute of Infectious Diseases, Higashimurayama-shi, Tokyo,3 Japan

Received 1 December 1999/Returned for modification 3 February 2000/Accepted 15 March 2000

We previously established a monoclonal antibody (MAb), designated H9, which reacts with the heat shock protein 60 (HSP60) homologue of Helicobacter pylori as well as with other bacterial and human HSP60s. To determine the importance of a cross-reactive epitope on H. pylori HSP60 in H. pylori immunopathogenesis, we performed (i) mapping of an epitope on H. pylori HSP60 recognized by the H9 MAb, (ii) analysis of immunoglobulin G responses of patients with or without H. pylori infection to its epitope region, and (iii) studies of the protective effect of immunization with its epitope region on H. pylori infection in mice. The epitope recognized by the H9 MAb was mapped to the sequence of amino acids 189 to 203 (VEGMQFDRGYLSPYF) on the H. pylori HSP60 molecule. It was confirmed that the synthesized peptide designated pH9 was recognized by the H9 MAb. Enzyme-linked immunosorbent assay analysis showed that patients with H. pylori infection (n = 349) had significantly lower titers of pH9 antibody than did uninfected patients (n = 200) (P < 0.001), but this was not the case with purified H. pylori HSP60 recombinant Escherichia coli GroEL, or recombinant human HSP60. In C57BL/6 mice immunized with the pH9 peptide with Freund's complete adjuvant (FCA), the number of H. pylori organisms colonizing the stomach was significantly lower than that in mice immunized with pCont plus FCA (P < 0.0001) or FCA only (P < 0.005). The results suggest that the immune response to the cross-reactive epitope (pH9 region) on H. pylori HSP60 is unique and might be associated with protection against H. pylori infection.

* Corresponding author. Mailing address: Department of Microbiology, Kyorin University School of Medicine, Mitaka, Tokyo 181-8611, Japan. Phone: 81-422-47-5511, ext. 3464. Fax: 81-422-44-7325. E-mail: hiro-ya{at}ta2.so-net.ne.jp.

Infection and Immunity, June 2000, p. 3448-3454, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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