Prokaryotic Expression Of Recombinant Human Interferon-λ1 And Studies Of Its Anti-virus Effects On Corneal Stromal Cells Of Rabbit

Objective: To obtain the recombinant protein of human interferon-λ1 (hIFN-λ1) and determine its potency. To study the anti-virus effects of hIFN-λ1 on Herpes Simplex virus (HSV) of rabbit corneal stromal cells.Methods: The interested gene of hIFN-λ1 was cloned into prokaryote expression vector pGEX-4T. The recombinant plasmid pGEX-4T-λ1 transfected in E. coli BL21. was induced by IPTG, the target protein was expressed and subsequently obtained by purifying inclusion body. Its potency was assayed by microtitration. The corneal stromal cells of rabbit were cultured by tissue explant. Microdose cytopathic method was used to demonstrate the anti-virus effects of hIFN-λ1,IFN-αstandard protein on rabbit corneal stromal cells.Result: The obtained hIFN-λ1 protein was assayed by SDS-PAGE and a protein electrophoresis strip with a relative molecular mass (Mr) of 33000 was detected. A potency of 125/2 IU/ml was determined by using microtitration technique after calibrated by IFN-αstandard protein. 32, 16, 8 TCID50 of HSV-I virus could be induced by 125/2, 125/4,125/8 IU/ml hIFN-λ1 and IFN-αin 48 hours. 16, 8 ,4 TCID50 of HSV-II virus could be induced by 125/2, 125/4,125/8 IU/ml IFN-α, 8, 4, 2 TCID50 of HSV-II virus could be induced by 125/2, 125/4, 125/8 IU/ml hIFN-λ1 in 48 hours.Conclusions: The hIFN-λ1 protein can be obtained after the recombinant plasmid pGEX-4T-λ1 transfected in E. coli BL21. was induced by IPTG. It has the anti-virus effects on HSV of rabbit corneal stromal cells. The anti-virus effects of hIFN-λ1,IFN-αon rabbit corneal stromal cells are similar. The anti-virus effects of hIFN-λ1 is one time weaker than that of IFN-αon rabbit corneal stromal cells...