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目的 探讨通过树突状细胞 (DC)提呈途径降低肿瘤抗原肽用量的可行性 ,了解热休克蛋白 70 (Hsp70 )和抗原肽修饰DC作用的特点。方法 以Hsp70于体外结合抗原肽 ,并于体外修饰DC ,检测修饰后DC的代谢活性及分泌的细胞因子 ;比较修饰后DC和Hsp70 H2 2肽对淋巴细胞的激活作用 ;检测激活的淋巴细胞对H2 2瘤细胞的细胞毒作用以及注射DC和Hsp70 H2 2肽对小鼠肿瘤的抑制作用。结果 Hsp70结合 0 .15 μgH2 2肽可使 2× 10 5DC成熟 ,4× 10 3 成熟DC可激活 2× 10 6淋巴细胞 ;Hsp70结合 0 .0 0 3μg肽修饰的DC或Hsp70结合 0 .15 μg肽直接刺激 ,可激活相同数量的淋巴细胞 ,产生同样的杀瘤效果。激活DC后再回输的治疗方式与直接注射Hsp70 肽复合物的治疗方式相比 ,抗原肽的用量可以降低 5 0倍。正常肝组织来源的混合肽结合Hsp70后 ,不能使DC成熟 ,亦不能通过DC途径活化脾淋巴细胞。结论 DC提呈抗原肽激活淋巴细胞的途径能够有效降低Hsp70 肿瘤抗原肽复合物使用剂量。正常细胞的混合肽不能通过Hsp70和DC提呈激活淋巴细胞 ,其诱发自身免疫应答的可能性极低
Objective To investigate the feasibility of reducing the amount of tumor antigen peptide by dendritic cell (DC) presentation and to understand the role of heat shock protein 70 (Hsp70) and antigen peptide modified DC. METHODS: Hsp70 was used to bind antigen peptides in vitro and the DCs were modified in vitro to detect the metabolic activity of DCs and the secreted cytokines. The activated DCs and Hsp70 H2 2 peptides were compared to activate lymphocytes. The activated lymphocyte pairs H2 2 tumor cells and the inhibitory effect of DC and Hsp70 H2 2 peptide on tumor in mice. Results 2 × 10 5 DC could be matured with Hsp70 in combination with 0.15 μg H 2 2 peptide and 2 × 10 6 with 4 × 10 3 mature DCs. Hsp70 combined with 0.030 μg peptide modified DC or Hsp70 combined with 0.15 μg Peptides directly stimulate the activation of the same number of lymphocytes, resulting in the same killing effect. Activation of DC and then return to the treatment of treatment compared with direct injection of Hsp70 peptide complex treatment, antigen peptide dosage can be reduced 50 times. The normal liver tissue-derived mixed peptide binds to Hsp70 and can not mature DCs or activate splenic lymphocytes by the DC pathway. Conclusions DC presenting antigen peptide activating lymphocyte pathway can effectively reduce the dosage of Hsp70 tumor antigen peptide complex. The mixed peptide of normal cells can not activate lymphocytes through activation of Hsp70 and DC, and the possibility of inducing autoimmune response is very low