IL-18 mutant polypeptide as well as preparation and usage
A mutant and expression vector technology, applied in the field of genetic engineering, can solve the problem of not being able to be converted into active
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Embodiment 1
[0042] Example 1 Preparation of IL-18 Mutant Polypeptide IL-18E121R
[0043] (1) Construction of IL-18 mutant expression vector
[0044] IL-18 is structurally similar to the IL-1 family, but not functionally the same. A detailed comparison of the amino acid sequences of IL-18 and IL-1 in 7 strains (human, rat, mouse, horse, cow, pig, dog) shows that IL-18 and IL-1 have high homology The sequence of the two suggests that the two have the same characteristics, while some sequences with low homology may be related to the unique function of IL-18, especially the highly conserved sites in various IL-18 lines that are different from IL-1 . Therefore, Glu121 was selected for mutation in the loop7 region of IL-18. Except for dog IL-18, this site is highly conserved among the six IL-18 strains, and this site has a negative charge, which may bind to the IL-18 receptor through an ionic bond. We mutated Glu121 to Arg, changed the charge properties of this site, and observed the effect...
Embodiment 2
[0066] Example 2 IL-18 Mutant Polypeptide E121R Treats Arthritis in Mice
[0067] (1) Establishment of collagen-induced arthritis (CIA) mouse model
[0068] DBA / 1 mice were immunized with type II natural bovine collagen to induce CIA. From 30 days after immunization, the mice were examined for the incidence of CIA every day. All mice were sacrificed 10 days after the onset of disease, and the degree of swelling of the first paw showing disease symptoms was measured with a precision caliper and the level of IL-6 was detected.
[0069] (2) Treatment of arthritis with IL-18 mutants
[0070] Treatment of DBA / 1 mice immunized with type II native bovine collagen was initiated at the first clinical sign of disease. IL-18E121R was intraperitoneally injected daily with two different concentrations of 1 and 0.1 mg / kg for 10 days. The mice in the control group were divided into two groups, one group was injected with IL-18 binding protein (IL-18BP) as a treatment control, and the othe...
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