[Primary study of recombinant immunotoxin IL-18-PE38 in treating rheumatoid arthritis].
OBJECTIVE
To establish a kind of gene therapy method of rheumatoid arthritis, to construct the interleukin-18-PE38 fusion gene expression vector and to explore the expression of the fusion gene in the chondrocytes and 3T3 cells.


METHODS
Interleukin-18-PE38 fusion gene was cleaved from plasmid PRKL459k-IL-18-PE38 by restriction enzyme digestion, then linked with vectors PsecTag2B and transformed into competence bacteria, positive clones were selected and confirmed by restrictive enzyme (EcoRI) digestion assay. The rearrangement plasmid PsecTag2B-IL-18-PE38 was transfected into 3T3 cells and mouse chondrocytes by liposome protocol (experimental group), null vector was used as negative control, and the transient expression was identified by fluorescence Restrictive enzymes digestion analysis revealed that the length of the interleukin-immunocytochemical assay.


RESULTS
18-PE38 fusion gene was 6000 bp. Fluorescence immunocytochemical method showed that fluorescence intensity of the experimental group is strong,while fluorescence intensity of the control group is weak.


CONCLUSIONS
The eukaryotic expression vector PsecTag2B-IL-18-PE38 is established successfully which can be expressed in the 3T3 cells and mouse chondrocytes. Our results lay a foundation for the further investigation for rheumatoid arthritis therapy.
Wu
Xia
X
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu Sichuan, 610041, PR China.
Yang
Chunlei
C
Zhang
Lin
L
D016475:3T3 Cells; D000818:Animals; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000628:therapy; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005098:Exotoxins / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D000922:Immunotoxins / Q000235:genetics / Q000378:metabolism*; D020382:Interleukin-18 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D005098:Exotoxins; D000922:Immunotoxins; D020382:Interleukin-18; D011993:Recombinant Fusion Proteins