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Infection and Immunity, August 2002, p. 4158-4164, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4158-4164.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Generation and Characterization of Human Monoclonal scFv Antibodies against Helicobacter pylori Antigens

Nicole Reiche, Andreas Jung, Thomas Brabletz, Tanja Vater, Thomas Kirchner, and Gerhard Faller^*

Institute of Pathology, University of Erlangen-Nuremberg, Erlangen, Germany

Received 10 January 2002/ Returned for modification 20 March 2002/ Accepted 20 May 2002

Infection with Helicobacter pylori is chronic despite a vigorous cellular and humoral immune response and causes severe pathology in some patients. In this study, phage display was used as a new approach in order to investigate the role of the host's humoral immune response in the pathogenesis of H. pylori gastritis. Human monoclonal single-chain Fv (scFv) antibody fragments against H. pylori cell lysate and the H. pylori urease were isolated from an immune phage display library, constructed from peripheral blood lymphocytes of an H. pylori-infected patient. After affinity selection, 23% of the clones tested showed binding activity against a lysate of the H. pylori Sydney strain in enzyme-linked immunosorbent assay (ELISA) and 9% bound the H. pylori urease. Further characterization by PCR-fingerprint analysis and sequencing revealed that two closely related H. pylori binders and one antiurease scFv could be isolated. The selected scFvs were highly specific as analyzed by ELISA and immunoblots using various bacterial lysates and recombinant proteins. Analysis of the humoral immune response following H. pylori infection using human monoclonal antibodies might contribute to a better understanding of the pathogenesis of the disease. Moreover, using immune phage display libraries, it might be possible for relevant epitopes of H. pylori antigens to be determined, which might be of use for vaccine development.

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* Corresponding author. Mailing address: Institute of Pathology, University of Erlangen-Nuremberg, Krankenhausstrasse 8-10, 91054 Erlangen, Germany. Phone: 49 9131 8522524. Fax: 49 9131 8524745. E-mail: gerhard.faller{at}patho.imed.uni-erlangen.de.

Editor: R. N. Moore

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Infection and Immunity, August 2002, p. 4158-4164, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4158-4164.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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