Maturation and Trafficking of Monocyte-Derived Dendritic Cells in Monkeys: Implications for Dendritic Cell-Based Vaccines

Abstract

Human dendritic cells (DC) have polarized responses to chemokines as a function of maturation state, but the effect of maturation on DC trafficking in vivo is not known. We have addressed this question in a highly relevant rhesus macaque model. We demonstrate that immature and CD40 ligand-matured monocyte-derived DC have characteristic phenotypic and functional differences in vitro. In particular, immature DC express CC chemokine receptor 5 (CCR5) and migrate in response to macrophage inflammatory protein-1α (MIP-1α), whereas mature DC switch expression to CCR7 and respond exclusively to MIP-3β and 6Ckine. Mature DC transduced to express a marker gene localized to lymph nodes after intradermal injection, constituting 1.5% of lymph node DC. In contrast, cutaneous DC transfected in situ via gene gun were detected in the draining lymph node at a 20-fold lower frequency. Unexpectedly, the state of maturation at the time of injection had no influence on the proportion of DC that localized to draining lymph nodes, as labeled immature and mature DC were detected in equal numbers. Immature DC that trafficked to lymph nodes underwent a significant up-regulation of CD86 expression indicative of spontaneous maturation. Moreover, immature DC exited completely from the dermis within 36 h of injection, whereas mature DC persisted in large numbers associated with a marked inflammatory infiltrate. We conclude that in vitro maturation is not a requirement for effective migration of DC in vivo and suggest that administration of Ag-loaded immature DC that undergo natural maturation following injection may be preferred for DC-based immunotherapy.

Pharmacokinetic Differences Between a T Cell-Tolerizing and a T Cell-Activating Peptide

“…Furthermore, adjuvants provide an Ag depot, allowing professional APCs to enter the vaccine site and collect the peptide. These Ag-loaded APCs then migrate to the lymph nodes and initiate T cell activation (22). This study presents evidence that the activating HPV16 E7 peptide persists in all organs longer than the tolerizing Ad5 E1A peptide as a result of a slow release of the HPV16 E7 peptide from the vaccine site (Fig.…”

The Duration of Signaling through CD40 Directs Biological Ability of Dendritic Cells to Induce Antitumor Immunity

“…It is demonstrated that CD40 stimulation induces the migration of DCs into secondary lymphoid tissues by up-regulating the expression of CCR7 (13,14). Indeed, CD40 stimulation for DCs is essential in the priming phase, since mice lacking CD40L showed defective priming ability accompanied by a disrupted migration of DCs for regional LNs (15).…”

Direct transfection and activation of human cutaneous dendritic cells

“…To effectively deliver gold particles to the cytoplasm of epidermal LC it was necessary to use a He discharge pressure of 800 psi, which is four times higher than the pressure commonly used to transfect mouse or nonhuman primate skin. 8,21 Even with the application of a high pressure, gold particles were mostly found in the epidermis and only a few beads were detected in the epidermal/dermal junction. Accordingly, transgenic proteins were expressed mainly in the epidermis.…”