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Objective:To construct recombinant E.coli LLO/OVA and investigate its tumor metastatic inhibition effect in B16 OVA melanoma challenged mice. Methods:Recombinant E.coli LLO/OVA was constructed and the expression of listeriolysin O(LLO) and ovalbumin(OVA) of the vaccine was determined by coomassie brilliant blue staining and western blotting. After 3 subcutaneous injections of E.coli LLO/OVA,the percentages of CD3+CD4+T,CD4+CD25+T,CD3+CD8+T and OVA257-264 SIINFEKL specific CD8+T cells were determined by flow cytomytry,and the tumor metastatic inhibition effect in B16 OVA melanoma challenged mice was observed. Results:Recombinant E.coli LLO/OVA was successfully constructed,and the expression of LLO and OVA of the vaccine was confirmed. After 3 subcutaneous injections of E.coli LLO/OVA and E.coli OVA in mice,the percentages of CD3+CD4+T,CD4+CD25+T and CD3+CD8+T cells were equivalent in the two groups of mice. However,there were significantly more OVA257-264 SIINFEKL specific CD8+T cells in E.coli LLO/OVA vaccinated mice than that in E.coli OVA vaccinated mice. The prophylactic E.coli LLO/OVA vaccination effectively prevented the tumor metastasis to lungs in B16 OVA melanoma challenged mice. Depletion of CD8+T cells significantly impaired the tumor inhibition effect of the vaccine in B16 OVA challenged mice. The therapeutic vaccination of E.coli LLO/OVA significantly prevented melanoma metastasis to lungs in B16 OVA challenged mice too. Conclusion:Recombinant E.coli LLO/OVA vaccination is highly effective in inhibiting murine malignant melanoma metastasis by promoting CD8+T cell immunity.
Objective: To construct recombinant E. coli LLO / OVA and investigate its tumor metastatic inhibition effect in B16 OVA melanoma challenged mice. Methods: Recombinant E. coli LLO / OVA was constructed and the expression of listeriolysin O (LLO) and ovalbumin (OVA) of the vaccine was determined by coomassie brilliant blue staining and western blotting. After 3 subcutaneous injections of E. coli LLO / OVA, the percentages of CD3 + CD4 + T, CD4 + CD25 + T, CD3 + CD8 + T and OVA257-264 SIINFEKL specific CD8 + T cells were determined by flow cytomytry, and the tumor metastatic inhibition effect in B16 OVA melanoma challenged mice was observed. Results: Recombinant E. coli LLO / OVA was successfully constructed, and the expression of LLO and OVA of the vaccine was confirmed. After 3 subcutaneous injections of E. coli LLO / OVA and E. coli OVA in mice, the percentages of CD3 + CD4 + T, CD4 + CD25 + T and CD3 + CD8 + T cells were equivalent in the two groups of However, there were significantly more OVA257-264 SIINFEKL specific CD8 + T cells i n E. coli LLO / OVA vaccinated mice than that in E. coli OVA vaccinated mice. The prophylactic E. coli LLO / OVA vaccination substantially prevented the tumor metastasis to lungs in B16 OVA melanoma challenged mice. Depletion of CD8 + T cells significantly impaired the tumor inhibition effect of the vaccine in B16 OVA challenged mice. The therapeutic vaccination of E. coli LLO / OVA significantly prevented melanoma metastasis to lungs in B16 OVA challenged mice too. Conclusion: Recombinant E. coli LLO / OVA vaccination is highly effective in inhibiting murine malignant melanoma metastasis by promoting CD8 + T cell immunity.