抗CD40抗体增强宫颈癌肽疫苗的治疗作用

目的：探讨抗CD40抗体对宫颈癌肽疫苗治疗作用的影响。方法将乳头状病毒结构性蛋白 E7表位肽（ E749－57，H－2Db限制性，CD8 T细胞表位）与Toll样受体7配体（Gardiquimod）组成疫苗，C57BL／6小鼠给予疫苗及抗CD40抗体免疫后取得外周血及脾组织CD8 T细胞，利用流式细胞仪分析特异性CD8 T细胞数量，Elisa检测CD8 T细胞与TC－1细胞（表达HPV E7蛋白的小鼠宫颈癌细胞）共孵育后干扰素（ INF－γ）表达水平。另取15只皮下荷瘤小鼠，监测免疫后肿瘤生长速度。结果与单用疫苗组相比，抗 CD40抗体明显增加小鼠外周血肿瘤特异性 CD8 T细胞数量（16．50±0．8185％ vs 9．747±1．834％，P＝0．0282），且内源性CD8 T细胞体外与TC－1细胞共孵育可以分泌INF－γ，体内显著抑制皮下肿瘤生长（ P＜0．001）。结论抗CD40抗体增强肽疫苗诱发的免疫应答，有潜力成为一种良好的佐剂。

Anti-CD40 Antibody Enhanced Therapeutic Effects of CervicaI Cancer Peptide Vaccine

OBJECTIVE To investigate the effects of anti-CD40 antibody on the cervical cancer peptide vac-cine.METHODS Peptide vaccine consists of Human Papillomavirus structural protein E7 epitope ( E749-57 , H-2Db restricted CD8 T cell epitope) and Toll like receptor 7 ligand (Gardiquimod).Blood and spleen tissues from C57BL/6 mice after peptide vaccine and anti-CD40 treatment were harvested for:1) analyzing the number of tumor specific CD8 T cells by flow cytometry,2 ) detecting INF-γsecretion after incubation with TC-1 tumor cells ( mice cervical cancer model expressing HPV E7 antigen).Therapeutic anti-tumor effects were investigated in 15 TC-1 bearingmice after different vaccination strategies.RESULTS Comparing with peptide vaccine group,peptide vaccineplus anti-CD40 significantly increased the number of tumor specific CD8 T cells in blood (16.50 ±0.8185% vs9.747 ±1.834 %,P =0.0282).Furthermore,endogenous tumor specific CD8 T cells can recognize TC-1 cells exvivo as increased IFN-γ secretion was detected after co-incubation,and dramatically inhibited tumor growth in vivo. CONCLUSION Anti-CD40 antibody increased the immune responses induced by peptide vaccine,is a goodcandidate for cancer therapeutic adjuvant.