In Vitro Multiparameter Assay Development Strategy toward Differentiating Macrophage Responses to Inhaled Medicines.
Although foamy macrophages (FMΦ) are commonly observed during nonclinical development of medicines for inhalation, there are no accepted criteria to differentiate adaptive from adverse FMΦ responses in drug safety studies. The purpose of this study was to develop a multiparameter in vitro assay strategy to differentiate and characterize different mechanisms of drug-induced FMΦ. Amiodarone, staurosporine, and poly(vinyl acetate) nanoparticles were used to induce distinct FMΦ phenotypes in J774A.1 cells, which were then compared with negative controls. Treated macrophages were evaluated for morphometry, lipid accumulation, gene expression, apoptosis, cell activation, and phagocytosis. Analysis of vacuolization (number/area vacuoles per cell) and phospholipid content revealed inducer-dependent distinctive patterns, which were confirmed by electron microscopy. In contrast to the other inducers, amiodarone increased vacuole size rather than number and resulted in phospholipid accumulation. No pronounced dysregulation of transcriptional activity or apoptosis was observed in response to sublethal concentrations of all inducers. Functionally, FMΦ induction did not affect macrophage activation by lipopolysaccharide, but it reduced phagocytic capacity, with different patterns of induction, severity, and resolution observed with the different inducers. An in vitro multiparameter assay strategy is reported that successfully differentiates and characterizes mechanisms leading to FMΦ induction by different types of agents.
#Medical University of Łódź, Muszyńskiego 1, 90-151 Łódź, Poland.
Ewelina
E
Hoffman
Abhinav
A
Kumar
Varsha
V
Kanabar
Matthew
M
Arno
Lucas
L
Preux
∇GE Healthcare Life Sciences, The Maynard Centre, Forest Farm Road, Whitchurch, Cardiff CF14 7YT, United Kingdom.
Val
V
Millar
Clive
C
Page
∥Division of Immunology, Infection and Inflammatory Diseases, Guy's Campus, King's College London, 15-16 Newcomen Street, London SE1 1UL, United Kingdom.
Helen
H
Collins
Ian
I
Mudway
Lea Ann
LA
Dailey
Ben
B
Forbes
D000280:Administration, Inhalation; D000638:Amiodarone / Q000008:administration & dosage / Q000494:pharmacology*; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D005487:Foam Cells / Q000187:drug effects*; D007928:Lethal Dose 50; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles; D011145:Polyvinyls / Q000008:administration & dosage / Q000494:pharmacology*; D019311:Staurosporine / Q000008:administration & dosage / Q000494:pharmacology*; D014617:Vacuoles / Q000187:drug effects / Q000378:metabolism
D011145:Polyvinyls; C013215:polyvinyl acetate; D019311:Staurosporine; D000638:Amiodarone
alveolar macrophage; foamy macrophage; morphometry; toxicology; vacuolation