Construction and preliminary investigation of a plasmid containing a novel immunotoxin DT390-IL-18 gene for the prevention of murine experimental autoimmune encephalomyelitis.
Experimental autoimmune encephalomyelitis (EAE) is a neuropathological animal model for multiple sclerosis. Antigen-presenting cells (APCs) expressing interleukin-18 receptor (IL-18R) were shown to be crucial in the beginning and progress of EAE. In this study we tested the effect of a novel recombinant immunotoxin targeting IL-18R-bearing APC for EAE prevention. The novel eukaryotic plasmid DT390-IL-18-SRalpha, encoding recombinant immunotoxin DT390-IL-18, was constructed. The immunotoxin consisted of IL-18 as the targeting moiety, and a truncated diphtheria toxin (DT) as the toxic moiety. Transfection assay and proliferation inhibition assay proved the immunotoxin could be expressed in vitro and was toxic to the activated mouse T cells. To evaluate the preventive effect of DT390-IL-18-SRalpha on EAE in vivo, cationic liposome-embedded DT390-IL-18-SRalpha was injected into the hind limbs of EAE mice. DT390-IL-18-SRalpha-treated mice showed a delayed manifestation of EAE and decreased symptoms compared to the mice treated with plasmid DT390-SRalpha or phosphate-buffered saline alone. A significant reduction in infiltrating inflammatory cells was detected in the brain tissues from immunotoxin-treated mice as compared with the controls by hematoxylin-eosin staining. This study suggested that the recombinant immunotoxin DT390-IL-18 could be expressed in vitro and in vivo, and prevented murine EAE effectively.
Jia
Jing
J
Department of Forensic Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Li
Hong
H
Tai
Shuhong
S
Lv
Meili
M
Liao
Miao
M
Yang
Zhihui
Z
Zhang
Beilei
B
Zhou
Bin
B
Zhang
Gengqian
G
Zhang
Lin
L
D000818:Animals; D004167:Diphtheria Toxin; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D005260:Female; D015316:Genetic Therapy; D007167:Immunotherapy; D000922:Immunotoxins / Q000627:therapeutic use*; D020382:Interleukin-18; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000627:therapeutic use*; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
D004167:Diphtheria Toxin; D000922:Immunotoxins; D020382:Interleukin-18; D011993:Recombinant Fusion Proteins