Immunological Study On Murine Bone Marrow-derived Dendritic Cells Infected With Murine Fibroblast Activation Protein α Recombinant Adenoviral Vectors Against Subcutaneous Transplantation Tumor Of Lewis Lung Carcinoma

Objectives:To transfer FAP αgene into murine dendritic cells by recombinant adenoviruswhich is encoding murine FAP α,and verify the expression of murine FAP αin murinedendritic cells and carcinoma associated fibroblasts.Then to investigate whether themurine bone marrow-derived DCs which infected with murine FAP αrecombinantadenovirus can inhibit or prevent the growth of lewis lung cancer or not.Methods:1. culture the C57Bl/6murine bone marrow-derived DCs and infect them withmurine FAP αrecombinant adenoviral vectors and murine EGFP recombinantadenoviral vectors.Then verify the expression of murine FAP αin murine DCs.2. Using lewis lung cancer cells to establish the lewis lung caner tumor modelsin C57Bl/6murine.3. Immunological study on cancer vaccine(murine dendritic cells infected withmurine FAP αrecombinant adenoviral vectors) against murine tumor model of lewislung cancer.In the prophylactic study,we injected female C57Bl/6murinesubcutaneously with dendritic cells,rAd-FAP α DCs or rAd-EGFP DCs(1time perweek,3times) at the right flanks.7days after the last inoculation,the murine wereinjected with lewis lung cancer cells in the right flank.Then the mice were observedfor56days.In the therapeutic study,we injected female C57Bl/6murinesubcutaneously with lewis lung cancer cells at the right flanks.8days after the lastinjection,the treatment murine were inoculated with dendritic cells,rAd-EGFPdendritic cells or rAd-FAP α dendritic cells(1time per week,3times) at the leftflanks.Then the size of the tumor were measured and recorded every2days(0.52xwidth2xlength).After the inoculation of lewis lung cancer cells,the micewere observed for survival rate in the next49days.Using standard methodology toassess the rate of survivors. Results：1.Murine Fibroblast activation Protein was detected in rAd-FAP α DCs andCarcinoma Association Fibroblast by western blot and was detected byimmunohistochemical method in cancer associate Fibroblasts.2.Western blot showed the expression of rAd-FAP α DCs mouse fibroblastactivation protein (FAP). Showed that rAd-FAP α DCs was prepared. Western blotCAF results were consistent with the results of immunohistochemistry, FAP inC57Bl/6mouse model of subcutaneous transplantation tumor in the tumormicroenvironment. In the prevention experiment, through56days of observation,vaccination with rAd FAP α DCs mice are resistant to Lewis significant lung cancereffect, resulted in prolonged survival of the mice, and vaccination with rAd EGFPDCs mice or DCs immunization of mice survival rate has significant difference(Kplan-Meier analysis, P<0.001)(Fig. C). While the rAd EGFP DCs mice wereinoculated subcutaneously with subcutaneous inoculation of DCs, mice, there was nosignificant difference in tumor volume (P>0.05). Application of rAd FAP α DCsvaccine can inhibit tumor growth, but can not eliminate tumor, the mice finally diedof cancer.3. In the experimental treatment, rAd FAP α DCs vaccine therapy cansignificantly inhibit the growth of Lewis lung cancer (as shown in figure D), in a24day observation period, subcutaneous inoculation of rAd FAP α DCs treatmentinhibited the growth of tumors in mice significantly (day twelfth, P&lt;0.05);and subcutaneous inoculation of rAd EGFP in DCs mice and DCs mice subcutaneousinoculation, there was no significant difference in tumor volume (P>0.05)Conclusion:rAd FAP α DCs vaccine can induce the mice to produce anti Lewis lung cancereffect, inhibit the growth of tumor. However, the antitumor immune response and cannot cause tumor eradication...