Evaluation of particle uptake in human blood monocyte-derived cells in vitro. Does phagocytosis activity of dendritic cells measure up with macrophages?
This work focuses on microparticles as potential antigen delivery systems to target professional antigen-presenting cells. Surface modified polystyrene microparticles were administered to human-derived macrophages (MPhis) and dendritic cells (DCs) in vitro to evaluate the phagocytosis activity of each cell type. To discriminate between internalised particles and those closely attached to the outside of the cells, particle internalisation was verified by confocal laser scanning microscopy. Especially positively charged particles tend to stick to the outer cell membrane and may lead to false positive results when measured by conventional microscopy. In contrast, fluorescence microscopy in combination with an extracellular fluorescence quenching agent (trypan blue) allows the unequivocal assessment of particle uptake for screening purposes. For this assay, the fluorescent label needs to be in direct contact to the quenching agent and cannot be localised inside the particle core. Different types of microparticles varying in size, surface-material and zeta potential resulted in vast differences regarding their uptake by MPhis and DCs as well as the maturation of DCs. Negatively-charged carboxylated and bovine serum albumin-coated particles were phagocytosed by MPhis to a relatively small extent. Interestingly, phagocytosis of these particles was still significantly lower in DCs while positively charged poly-L-lysine (PLL) coated particles induced high phagocytosis activity in both cell types. By comparing our results with literature data, we conclude that phagocytosis activity of DCs and MPhis largely depends on particle size and surface charge and is also influenced by the character of bulk and coating material. PLL can be directed to DCs and MPhis with comparable efficiency and, in addition, induce maturation of DCs.
Thiele
L
L
Department of Applied BioSciences, Swiss Federal Institute of Technology Zurich (ETH), Winterthurerstrasse 190, CH-8057, Zurich, Switzerland.
Rothen-Rutishauser
B
B
Jilek
S
S
Wunderli-Allenspach
H
H
Merkle
H P
HP
Walter
E
E
D015703:Antigens, CD; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000502:physiology*; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D008264:Macrophages / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000032:analysis; D018613:Microscopy, Confocal; D010316:Particle Size; D010587:Phagocytosis; D000097802:CD83 Antigen
D015703:Antigens, CD; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D000097802:CD83 Antigen