Title: Construction and expression of converting enzyme coexpression vector for murine interleukin-18 and interleukin-1β
Abstract: To prepare the tumor vaccine coexpressing precursor interleukin-18 (proIL-18) and IL-1β converting enzyme (ICE) gene, proIL-18 and ICE genes were cloned from murine splenocytes, and their coexpression vector pIRES-proIL-18-ICE was constructed. After confirming the sequence of the insert to be correct, the recombinant vector was transfected into L1210, a murine lymphoid leukemia cell line, by using the Lipofectamine reagent. The stable expression transfectants were selected by G418. RT-PCR analysis of transfectant cells and ELISA assay for IL-18 in the culture supernatants indicated that the transfectant tumor cells could express and secret mature IL-18. The IL-18 secreted to culture supernatants apparently stimulated IFN-γ production in ConA primed murine splenocytes, suggesting IL-18 secreted from transfectants retained its bioactivity. In addition, the cell growth curve and cell cycle test showed that transfection of pIRES, pIRES-proIL-18, pIRES-ICE and pIRES-proIL-18-ICE have little influence on the growth and proliferation of L1210 cells. The successful production of IL-18 leukemia vaccine indicates that, similar to physiological state, coexpression of proIL-18 and ICE in the same tumor cell using pIRES vector can result in efficient cleavage and secretion of bioactive mature IL-18. The IL-18 leukemia vaccine can be used in future study to assess its efficacy of anti-leukemic immunity in a murine model.
Publication Year: 2005
Publication Date: 2005-01-01
Language: en
Type: article
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