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Department of Cell and Molecular Biology, Section for Immunology, Lund University, Lund, Sweden
Salmonella typhimurium is an intracellular bacterium
that replicates in the spleen and mesenteric lymph nodes (MLN) of
orally infected mice. However, little is known about the Ag
presentation and cytokine production capacity of dendritic cells (DC),
particularly CD8
+,
CD8-CD4-, and
CD8-CD4+ DC, from these organs in response
to Salmonella. Infection of purified splenic DC with
S. typhimiurium expressing green fluorescent protein
(GFP) and OVA revealed that all three splenic DC subsets internalize
bacteria, and splenic as well as MLN DC process
Salmonella for peptide presentation. Furthermore,
presentation of Salmonella Ags on MHC-I and MHC-II was
evident in both CD8+ and CD8- splenic DC
subsets. Direct ex vivo analysis of splenic DC from mice infected with
GFP-expressing Salmonella showed that all three subsets
harbored bacteria, and splenic DC purified from mice given
Salmonella-expressing OVA presented OVA-derived peptides
on MHC-I and MHC-II. Cytokine production analyzed by intracellular
staining of splenic DC infected with GFP-expressing
Salmonella revealed that TNF- was produced by a large
percentage of CD8- DC, while only a minor proportion of
CD8+ DC produced this cytokine following bacterial
exposure. In contrast, the greatest number of IL-12p40-producing DC
were among CD8+ DC. Experiments inhibiting bacterial
uptake by cytochalasin D as well as use of a Transwell system revealed
that bacterial contact, but not internalization, was required for
cytokine production. Thus, DC in sites of Salmonella
replication and T cell activation, spleen and MLN, respond to bacterial
encounter by Ag presentation and produce cytokines in a subset-specific
fashion.
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