Soluble and Microparticle‐Based Delivery of TLR4 and TLR9 Agonists Differentially Modulate 3D Chemotaxis of Bone Marrow‐Derived Dendritic Cells

Abstract Vaccines are commonly administered subcutaneously or intramuscularly, and local immune cells, notably dendritic cells (DCs), play a significant role in transporting vaccine antigens and adjuvants to draining lymph nodes. Here, it is compared how soluble and biomaterial‐mediated delivery of Toll‐like receptor (TLR)‐targeted adjuvants, monophosphoryl lipid A (MPLA, TLR4 ligand) and 5'‐C‐phosphate‐G‐3' DNA (CpG DNA, TLR9 ligand), modulate 3D chemotaxis of bone marrow‐derived dendritic cells (BMDCs) toward lymphatic chemokine gradients. Within microfluidic devices containing 3D collagen‐based matrices to mimic tissue conditions, soluble MPLA increases BMDC chemotaxis toward gradients of CCL19 and CCL21, while soluble CpG has no effect. Delivering CpG on poly(lactic‐ co ‐glycolic) acid microparticles (MPs) enhances BMDC chemotaxis compared to MPLA‐encapsulated MPs, and when co‐delivered, MPLA and CpG do not synergistically enhance BMDC migration. It is concluded that supplementing granulocyte‐macrophage colony stimulating factor‐derived BMDC culture with interleukin‐4 is necessary to induce CCR7 expression and chemotaxis of BMDCs. Different cell subsets in BMDC culture upregulate CCR7 in response to soluble versus biomaterial‐loaded MPLA and CpG, and CCR7 expression does not consistently correlate with functional migration. The results show both adjuvant type and delivery method influence chemotaxis of DCs, and these findings uncover new directions for the rational design of vaccine formulations.