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- Dendritic Cells to Enhance Presentation of a Tumor Peptide In Vivo1
Departments of
*
Experimental Medicine and
Pharmacology, University of Perugia, Perugia, Italy
Previous work has shown that a significant proportion of murine
splenic dendritic cells (DC) express a high affinity receptor for
IL-12, thus accounting for the adjuvanticity of the cytokine when DBA/2
mice are transferred with syngeneic DC exposed in vitro to rIL-12 and
an otherwise poorly immunogenic tumor peptide. In DBA/2 mice, splenic
DC consist of 90–95% CD8- and 5–10% CD8+
cells. To detect any difference in IL-12 responsiveness among
phenotypically distinct DC subtypes, enriched CD8- (>99%
pure) and CD8+ (
80% pure) populations of DC from DBA/2
spleens were assayed for APC function in vivo following exposure to
rIL-12 and tumor peptide in vitro. Unlike unfractionated DC, the
CD8- fraction was capable of effective presentation of the
peptide even when the cells had not been pretreated with IL-12 before
peptide pulsing. The addition of as few as 3% CD8+ cells
during pulsing blocked in vivo priming by the CD8-
fraction. However, pretreatment of CD8- DC with IL-12
before cell mixing and peptide pulsing ablated the inhibitory effect of
the CD8+ fraction. CD8-, but not
CD8+, DC showed significant message expression for the ß1
and ß2 subunits of the IL-12 receptor. These data suggest that a
minority population of CD8+ DC, which appeared to secrete
IL-10 in vitro, negatively regulates the induction of T cell reactivity
by peptide-loaded CD8- DC in DBA/2 mice. However, the
CD8- fraction can be primed by IL-12 to overcome the
inhibitory effect of the CD8+
subtype.
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