功能化氧化石墨烯GCF的制备及其增强OVA的免疫原性评价  

Preparation of functional oxide graphene and evaluation of its immunogenicity in enhancing OVA

作　　者：宣德春 尹文竹 王海燕[2] 周明旭 邓碧华[2] 马芳 卢宇 张金秋[1,2,3,4] XUAN Dechun;YIN Wenzhu;WANG Haiyan;ZHOU Mingxu;DENG Bihua;MA Fang;LU Yu;ZHANG Jinqiu(School of Pharmacy,Jiangsu University,Zhenjiang,Jiangsu 212013,China;National Research Center of Engineering and Technology for Veterinary Biologicals,Institute of Veterinary Immunology&Engineering,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou,Jiangsu 225009,China;Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base,Ministry of Science and Technology,Nanjing 210014,China)

机构地区：[1]江苏大学药学院,江苏镇江212013 [2]江苏省农业科学院动物免疫工程研究所国家兽用生物制品工程技术研究中心,江苏南京210014 [3]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [4]江苏省食品质量安全重点实验室/省部共建国家重点实验室培育基地,江苏南京210014

出　　处：《中国兽医学报》2023年第3期562-570,共9页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32172895,31772701);江苏省农业科技自主创新基金资助项目(CX(17)3027)。

摘　　要：为制备高效负载、递送和缓释抗原的新型功能化氧化石墨烯衍生材料(GCF)作为佐剂,探索该纳米佐剂对OVA免疫原性影响。以氧化石墨烯(graphene oxide,GO)为母体,采用EDC/NHS共价嫁接壳聚糖(chitosan,CS),并静电吸附泊洛沙姆407(poloxamer,F127)制备出亲水性的氧化石墨烯衍生材料GO-CS-F127(简称GCF)。利用XRD、UV-vis、FI-IR、Zeta电位等多种方法对GCF进行理化特性表征,确定材料的形貌、成分及电荷分布。通过BCA法检测GCF对OVA的最大吸附量及释放率,CCK-8法鉴定GCF新材料在体外安全性评价,ELSIA方法评价小鼠血清中特异性抗体水平,并采集小鼠重要组织制备切片以观察病理变化。结果显示,GCF呈片层结构,常温放置30 d仍能稳定存放,带有17.9 mV的正电荷,能快速吸附抗原,其最大吸附量能达到1.9 g/g,且对抗原具有显著的缓释作用。GCF质量浓度在100 mg/L以下,细胞存活率达85%以上,且对心、脾、肾等组织无损伤,具有良好的生物相容性。与单独OVA组及PBS组相比,发现GCF能显著提高小鼠OVA特异性IgG抗体水平。结果表明,以功能化氧化石墨烯GCF作为佐剂,可有效提高抗原的免疫原性。Hydrophilic oxide graphene(GO)complex nanomaterials(GCF)were fabricated using GO as framework for covalent grafting of chitosan(CS)with EDC/NHS and then electrostatic adsorption of poloxamer 407(F127).The GCF was characterized by XRD,UV-vis,FI-IR and Zeta potential detection.Due to its 2D laminar structure,good biocompatibility and a positive charge of 17.9 mV,the GCF can efficiently load,deliver and slowly release antigens.Using OVA as the mode antigen,GCF can quickly absorb OVA with a maximum adsorption amount of 1.9 g/g and has a significant slow-release effect on the antigen.The result of CCK8 assay showed when the concentration of GCF was below 100mg/L,the cell survival rate was more than 85%.Meanwhile,the H&E results indicated that there was no damage to heart,spleen,kidney and other tissues.As a kind of novel nano-adjuvants,GCF could obviously improve the level of OVA-specific IgG antibodies in mice.Our research results suggested GCF could be used as an adjuvant and effectively improve the immune response of vaccines with good biocompatibility.

关 键 词：氧化石墨烯 壳聚糖 共价键 佐剂 疫苗 

分 类 号：R392[医药卫生—免疫学]