Prevention of murine experimental autoimmune encephalomyelitis by in vivo expression of a novel recombinant immunotoxin DT390-RANTES.
Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediated autoimmune disease. Chemokine receptor CCR5 has been shown to be essential for the T-cell recruitment to the inflammatory site in EAE. In this study, we assumed that an immunotoxin directed at CCR5+ cells would be able to reduce the disease activity of EAE. A recombinant immunotoxin, DT390-RANTES-SRalpha, was constructed in an eukaryotic cell expression plasmid consisting of regulated on activation normal T cells expressed and secreted (RANTES) as the targeting moiety and DT390 as the toxic moiety. DT390-RANTES was expressed in vitro and was highly toxic to activated mouse T cells with the inhibitory concentration 50 at 0.18 ng/ml. To evaluate whether DT390-RANTES was effective in preventing EAE, C57BL/6 mice were immunized with myelin basic protein, emulsified with complete Freund's adjuvant and were treated by injecting cationic liposome-embedded plasmid DNA into the muscle of hind limbs. Mice treated with DT390-RANTES-SRalpha developed a much milder EAE compared to mice treated with phosphate-buffered saline or the empty plasmid DNA. Much less CCR5+-infiltrating cells were found in the central nervous system in DT390-RANTES-SRalpha-treated mice than in the control mice. This study indicates that recombinant immunotoxin can be expressed in vivo, and targeting CCR5 can attenuate the disease activity of EAE in mice.
Jia
Y
Y
West China School of Preclinical and Forensic Medicine, Sichuan University, Sichuan, People's Republic of China.
Li
H
H
Chen
W
W
Li
M
M
Lv
M
M
Feng
P
P
Hu
H
H
Zhang
L
L
D016475:3T3 Cells; D000818:Animals; D018946:Chemokine CCL5 / Q000235:genetics / Q000276:immunology; D002634:Chemotaxis, Leukocyte; D004247:DNA / Q000008:administration & dosage / Q000378:metabolism; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology / Q000517:prevention & control*; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007114:Immunization; D007167:Immunotherapy / Q000379:methods*; D000922:Immunotoxins / Q000008:administration & dosage* / Q000235:genetics; D007273:Injections, Intramuscular; D051379:Mice; D008810:Mice, Inbred C57BL; D009103:Multiple Sclerosis / Q000276:immunology / Q000517:prevention & control; D018482:Muscle, Skeletal / Q000276:immunology*; D004676:Myelin Basic Protein / Q000008:administration & dosage / Q000378:metabolism; D019713:Receptors, CCR5 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection / Q000379:methods
D018946:Chemokine CCL5; C518733:DT390-RANTES-SRalpha; D000922:Immunotoxins; D004676:Myelin Basic Protein; D019713:Receptors, CCR5; D011993:Recombinant Fusion Proteins; D004247:DNA