以Candin为佐剂的人乳头瘤病毒多肽治疗疫苗刺激朗格汉斯细胞分泌IL-12的研究

Secretion of IL-12 by Langerhans cells stimulated with a human papillomavirus peptide-based therapeutic vaccine containing Candin as an adjuvant

目的探讨佐剂Candin及包含佐剂Candin和HPV多肽的疫苗对朗格汉斯细胞(langerhans cells,LCs)分泌IL-12的影响。方法采用淋巴细胞分离液分离人外周血PBMCs,采用磁珠分选系统分选CD14+细胞,经细胞因子rhGM-CSF(1 000IU/ml)、rhIL-4(1 000IUs)、TGF-β1(10ng/ml)诱导后获得LCs,采用流式细胞术鉴定。用佐剂Candin(50、100、150μl/ml)、包含佐剂Candin(50、100、150μl/ml)和HPV16E6多肽(10μmol/ml)疫苗刺激LCs 24h,ELISA法检测IL-12p70水平。结果诱导培养7d的细胞高表达LCs特征性的表面分子Langerin。佐剂Candin及HPV多肽疫苗均能不同程度刺激LCs分泌IL-12p70。佐剂Candin刺激LCs分泌IL-12p70最高达175ng/ml,HPV多肽疫苗刺激组最高达298ng/ml。含有Candin 100μl/ml的HPV多肽疫苗刺激LCs分泌IL-12p70水平较相应剂量的单独佐剂Candin组高。结论佐剂Candin及HPV多肽疫苗均能刺激LCs分泌IL-12。HPV多肽疫苗刺激LCs分泌IL-12的能力较单独佐剂Candin更强。

Objective To investigate the effect of the adjuvant Candin and a HPV peptide-based therapeutic vaccine containing Candin and HPV peptides on the secretion of IL-12 by Langerhans cells. Methods PBMCs were separated and purified using Ficoll gradient centrifugation and CD14＋cells were separated using a CD14 microbead kit.LCs were induced using rhGM-CSF（1,000IU/ml）,rhIL-4（1,000IUs）,and TGF-β1（10ng/ml）,and LCs were identified with FCM.LCs were stimulated with Candin（50,100,and 150μl/ml）and a HPV peptide-based vaccine containing Candin（50,100,150μl/ml）and HPV16E6peptide（10μmol/ml）for 24 h.IL-12p70 secreted by LCs was measured with ELISA. Results The LC-specific molecule langerin was highly expressed in induced cells.IL-12p70 was secreted by LCs stimulated with Candin or the vaccine.IL-12p70 reached 175ng/ml in LCs stimulated with Candin and 298ng/ml in LCs stimulated with the vaccine.More IL-12p70 was secreted by LCs stimulated with the vaccine containing Candin 100μl/ml compared to LCs stimulated with the corresponding amount of Candin. Conclusion Both Candin and the vaccine have the ability to stimulate LCs to secrete IL-12 and the vaccine is better able to stimulate those cells than Candin.