摘要
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,CD8T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CD8T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。
OBJECTIVE To investigate the effects of anti-CD40 antibody on the cervical cancer peptide vaccine. 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 C57 BL /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-γ secretion 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 bearing mice after different vaccination strategies. RESULTS Comparing with peptide vaccine group,peptide vaccine plus 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 ex vivo 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 good candidate for cancer therapeutic adjuvant.
出处
《海峡药学》
2016年第3期37-39,共3页
Strait Pharmaceutical Journal
