Novel Therapy for Atherosclerosis Using Recombinant Immunotoxin Against Folate Receptor β–Expressing Macrophages

Abstract

BackgroundFolate receptor β (FRβ) is induced during macrophage activation. A recombinant immunotoxin consisting of the truncatedPseudomonasexotoxin A (PE38) conjugated to an anti‐FRβ antibody (anti–FRβ‐PE38) has been reported to kill activated macrophages in inflammatory diseases. To elucidate the effect of an immunotoxin targetingFRβ on atherosclerosis, we determined the presence ofFRβ‐expressing macrophages in atherosclerotic lesions and administered theFRβ immunotoxin in apolipoprotein E–deficient mice.Methods and ResultsTheFRβ‐expressing macrophages were observed in atherosclerotic lesions of apolipoprotein E–deficient mice. At 15 or 35 weeks of age, the apolipoprotein E–deficient mice were divided into 3 groups and were intravenously administered 0.1 mg/kg of anti–FRβ‐PE38 (immunotoxin group), 0.1 mg/kg of PE38 (toxin group), or 0.1 mL of saline (control group) every 3 days, for a total of 5 times for each age group. The mice were analyzed at 21 or 41 weeks of age. Treatment with the immunotoxin resulted in 31% and 22% reductions in atherosclerotic lesions of the 21‐ and 41‐week‐old mice, respectively (P<0.05). Administration of immunotoxin reduced the numbers ofFRβ‐ and tumor necrosis factor‐α–expressing macrophages, reduced cell proliferation, and increased the number of apoptotic cells (P<0.05). Real‐time polymerase chain reaction demonstrated that the expression ofFRβ and tumor necrosis factor‐αmRNAwas significantly decreased in the immunotoxin group (P<0.05).ConclusionsThese results suggest thatFRβ‐expressing macrophages exist in the atherosclerotic lesions of apolipoprotein E–deficient mice and thatFRβ immunotoxin administration reduces the progression of atherosclerotic lesions in younger and older individuals. The recombinantFRβ immunotoxin targeting activated macrophages could provide a novel therapeutic tool for atherosclerosis.