Nature Biotechnology
20, 702 - 706 (2002)
doi:10.1038/nbt0702-702
In situ delivery of passive immunity by lactobacilli producing
single-chain antibodiesCarina Krüger1, Yanzhong Hu1, Qiang Pan1, Harold Marcotte1, Anna Hultberg1, Dipu Delwar1, Philip J. van Dalen2, Peter H. Pouwels2, 3, Rob J. Leer2, 3, Charles G. Kelly4, Craig van Dollenweerd4, Julian K. Ma4
& Lennart Hammarström11
Center for Oral Biology, Karolinska Institutet at
NOVUM and Division of Clinical Immunology, Karolinska Institutet at Huddinge
Hospital, SE-141 86 Huddinge,
Sweden. 2
TNO Prevention and Health, Division of Immunology
and Infectious Disease, PO Box 2215, 2301 CE
Leiden, The Netherlands. 3
TNO Voeding, Department of Applied Microbiology
and Gene Technology, Postbox 370, 3600 AJ, Zeist,
The Netherlands. 4
Department of Oral Medicine and Pathology, King's
College London, Guy's Hospital, London Bridge,
London, UK.
Correspondence should be addressed to Lennart Hammarström lennart.hammarstrom@biosci.ki.seLactobacilli 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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