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In situ delivery of passive immunity by lactobacilli producing single-chain antibodies

Nature Biotechnology volume 20, pages 702–706 (2002)Cite this article

Abstract

Lactobacilli have previously been used to deliver vaccine components for active immunization in vivo. Vectors encoding a single-chain Fv (scFv) antibody fragment, which recognizes the streptococcal antigen I/II (SAI/II) adhesion molecule of Streptococcus mutans, were constructed and expressed in Lactobacillus zeae (American Type Culture Collection (ATCC) 393). The scFv antibody fragments secreted into the supernatant or expressed on the surface of the bacteria showed binding activity against SAI/II in enzyme-linked immunosorbent assay (ELISA), and surface scFv-expressing lactobacilli agglutinated SAI/II-expressing S. mutans in vitro without affecting the corresponding SAI/II knockout strain. Lactobacilli expressing the scFv fragment fused to an E-tag were visualized by scanning electron microscopy (SEM) using beads coated with a monoclonal anti-E-tag antibody, and they bound directly to beads coated with SAI/II. After administration of scFv-expressing bacteria to a rat model of dental caries development, S. mutans bacteria counts and caries scores were markedly reduced. As lactobacilli are generally regarded as safe (GRAS) microorganisms, this approach may be of considerable commercial interest for in vivo immunotherapy.

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Figure 1: Map of Lactobacillus expression vectors.
Figure 2: Detection of the scFv expressed by L. zeae by immunoblotting and fluorescence-activated cell sorting (FACS).
Figure 3: Adherence of transformed lactobacilli to magnetic beads coated with E-tag antibodies or SAI/II.

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Acknowledgements

This work was supported by the Swedish Medical Research Council and by the Foundation for Preventive Caries Research (Patentmedelsfonden).

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Authors and Affiliations

  1. Karolinska Institutet at NOVUM and Division of Clinical Immunology, Center for Oral Biology, Karolinska Institutet at Huddinge Hospital, Huddinge, SE-141 86, Sweden

    Carina Krüger, Yanzhong Hu, Qiang Pan, Harold Marcotte, Anna Hultberg, Dipu Delwar & Lennart Hammarström

  2. Division of Immunology Infectious Disease, TNO Prevention and Health, PO Box 2215, Leiden, 2301 CE, The Netherlands

    Philip J. van Dalen, Peter H. Pouwels & Rob J. Leer

  3. Department of Applied Microbiology Gene Technology, TNO Voeding, Postbox 370, Zeist, 3600 AJ, The Netherlands

    Peter H. Pouwels & Rob J. Leer

  4. Department of Oral Medicine and Pathology, King's College London, Guy's Hospital, London Bridge, London, UK

    Charles G. Kelly, Craig van Dollenweerd & Julian K. Ma

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Correspondence to Lennart Hammarström.

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Krüger, C., Hu, Y., Pan, Q. et al. In situ delivery of passive immunity by lactobacilli producing single-chain antibodies. Nat Biotechnol 20, 702–706 (2002). https://doi.org/10.1038/nbt0702-702

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