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*
Laboratory of Immunology, Istituto Dermopatico dell’Immacolata, Instituto di Ricovero e Cura a Carattere Scientifico, Rome, Italy;
Section of Microbiology, Department of Molecular Biology, University of Siena, Siena, Italy; and
Department of Biotechnology and Bioscience, University of Milan-Bicocca, Milan, Italy
Recombinant Streptococcus gordonii expressing on the
surface the C-fragment of tetanus toxin was tested as an Ag delivery
system for human monocyte-derived dendritic cells (DCs). DCs incubated
with recombinant S. gordonii were much more efficient
than DCs pulsed with soluble C-fragment of tetanus toxin at stimulating
specific CD4+ T cells as determined by cell proliferation
and IFN-
release. Compared with DCs treated with soluble Ag, DCs fed
with recombinant bacteria required 102- to
103-fold less Ag and were at least 102 times
more effective on a per-cell basis for activating specific T cells.
S. gordonii was internalized in DCs by conventional
phagocytosis, and cytochalasin D inhibited presentation of
bacteria-associated Ag, but not of soluble Ag, suggesting that
phagocytosis was required for proper delivery of recombinant Ag.
Bacteria were also very potent inducers of DC maturation, although they
enhanced the capacity of DCs to activate specific CD4+ T
cells at concentrations that did not stimulate DC maturation. In
particular, S. gordonii dose-dependently up-regulated
expression of membrane molecules (MHC I and II, CD80, CD86, CD54, CD40,
CD83) and reduced both phagocytic and endocytic activities.
Furthermore, bacteria promoted in a dose-dependent manner DC release of
cytokines (IL-6, TNF-
, IL-1ß, IL-12, TGF-ß, and IL-10) and of
the chemokines IL-8, RANTES, IFN--inducible protein-10, and monokine
induced by IFN-. Thus, recombinant Gram-positive bacteria appear a
powerful tool for vaccine design due to their extremely high capacity
to deliver Ags into DCs, as well as induce DC maturation and secretion
of T cell chemoattractans.
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