Specific Anti-Hepatoma Immunity Induced By DC Stimulated By HSP70-H22 Peptide Complexes And SCD40L

Objective:To study the specific anti-tumor effect of dendritic cells(DCs) modified by Hsp70-tumor peptide complexes and sCD40L in vitro and in vivo.Method:1. The murine marrow cells were cultured for 7days,then randomized to 4 groups, and stimulated respectively for 24 h by no stimulus,only Hsp70-H22 peptide complexes,sCD40L and Hsp70-H22 peptide complexes combined with sCD40L. IL-12 and IL-10 of supernatant were tested by ELISA assay ,CD40 and CD80 of DCs were tested by FACS and the stimulus index (SI)was tested by MLR . The ability of spleen lymphocytes activated by DCs to H22 cells in vitro was investigated by MTT.2. The models of murine hepatoma was established, then completely randomrized to 5 groups(Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ),and interfered respectively by normal saline and DCs respectively drived from the four stimulated groups above, day 21th,the murines were sacrificed .The weight of tumor was measured . The level of IL-10 and INF-γin blood serum was tested by ELISA assay.Result:1. Compared with the group stimulated by no stimulus , the level of IL-10 of all the other groups decreased remarkably, and the level of CD40,CD80,IL-12,SI and the ratio of spleen lymphocytes activated by DCs to H22 cells in vitro of the groups stimulated respectively by only Hsp70-H22 peptide complexes and Hsp70-H22 peptide complexes combined with sCD40L increased significantly, the differences have statistical significance(P<0.05); all the differences of SI and the ratio of spleen lymphocytes activated by DCs to H22 cells in vitro between the group stimulated by sCD40L and the group stimulated by no stimulus have no statistical significance(P>0.05).2. Compared with the group stimulated by sCD40L,the level of IL-10 of group stimulated by Hsp70-H22 peptide complexes decreased remarkably; SI and the ratio of spleen lymphocytes activated by DCs to H22 cells in vitro increased remarkably, all the differences have statistical significance(P<0.05),but the differences of CD40,CD80,IL-12 have no statistical significance(P>0.05). The level of IL-10 of group stimulated by Hsp70-H22 peptide complexes combined with sCD40L decreased remarkably, and the level of CD40,CD80,IL-12,SI and the ratio of spleen lymphocytes activated by DCs to H22 cells in vitro increased significantly, the differences have statistical significance(P<0.05).3. Compared with the group stimulated by Hsp70-H22 peptide complexes, the level of IL-10 of group stimulated by Hsp70-H22 peptide complexes combined with sCD40L decreased remarkably, the level of CD40,CD80,IL-12,SI and the ratio of spleen lymphocytes activated by DCs to H22 cells in vitro increased significantly,all the differences have statistical significance(P<0.05).4. Compared with groupⅠ, the level of IL-10 and the weight of tumor of groupsⅣandⅤdecreased markedly, and the level of INF-γincreased remarkably, all the differences have statistical significance(P<0.05);all the results of groupsⅡandⅢhave no statistical significance(P>0.05)5. Compared with groupⅡ, the level of IL-10 and the weight of tumor of groupsⅣandⅤdecreased markedly, and the level of INF-γincreased remarkably, all the differences have statistical significance(P<0.05);all the results of groupsⅢhave on statistical significance(P>0.05).6. Compared with groupⅢ, the level of IL-10 and the weight of tumor of groupsⅣandⅤdecreased markedly, and the level of INF-γincreased remarkably, all the differences have statistical significance(P<0.05).7. Compared with groupⅣ, the level of IL-10 and the weight of tumor of groupⅤdecreased markedly, and the level of INF-γincreased remarkably, all the differences have statistical significance(P<0.05).Conclusion:DCs modified by Hsp70-tumor peptide complexes and sCD40L which can be activated sufficiently are able to induce powerful specific antitumor immunity and remarkably killing-effect to the H22 cell, resulting in tumor growth being significantly inhibited.