健康人外周血白念珠菌烯醇化酶抗原特异性T细胞检测  

Detection and Analysis of Candida Albicans Enolase Antigen-Specific T Cells in Heathy Individuals by ELISPOT Assay

作　　者：蒋曦旻 张斌[2] 史利宁 商秀娟 张彩云 王静瑜 胡毓安[3] JIANG Xi-min;ZHANG Bin;SHI Li-ning;SHANG Xiu-juan;ZHANG Cai-yun;WANG Jing-yu;HU Yu-an(a. IT Center;lb. Department of Central Laboratory Medicine, Nanjing General Hospital, PLA , Nanjing 210002, China;2. Department of Pharmacy, the Affiliated Drumtower Hospital of Nanjing University Medical School, Nanjing 210008,Chin)

机构地区：[1]中国人民解放军南京总医院医学信息科,南京210008 [2]南京大学医学院附属鼓楼医院药学部,南京210008 [3]中国人民解放军南京总医院临床中心实验科,南京210002

出　　处：《现代检验医学杂志》2018年第1期10-13,共4页Journal of Modern Laboratory Medicine

基　　金：南京军区医药卫生科研基金课题面上项目(15MS125);国家自然科学基金青年科学基金项目(81302536)

摘　　要：目的研究白念珠菌免疫优势抗原烯醇化酶(Enolase,Eno)的细胞免疫应答类型。方法以白念珠菌重组抗原Eno为刺激物,细胞刺激剂PHA为阳性对照,用酶联免疫斑点技术(enzyme linked immunospot assay,ELISPOT)检测25例健康人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)分泌干扰素γ(interferonγ,IFN-γ)、白细胞介素-4(interleukin-4,IL-4)和白细胞介素-17A(interleukin-17,IL-17A)的阳性斑点形成细胞(spot forming cell,SFC)频数。结果Eno刺激后,25例健康人分泌IFN-γ,IL-4和IL-17A的Eno抗原特异性T淋巴细胞频数分别为14.00(8.50,39.00)、0(0,0)和2(1,4.50)个,三者间差异具有统计学意义(均P<0.05)。IFN-γ,IL-4和IL-17A的应答率分别为100%(25/25),4.00%(1/25)和88.00%(22/25),三者之间差异具有统计学意义(均P<0.05)。Eno刺激后的强IFN-γ(SFC≥20)应答率为40.00%(10/25),无受试者产生强IL-17A和强IL-4应答。受试者对Eno的应答模式以同时产生Th1和Th17型应答为主,占84.00%(21/25);3例研究对象仅产生Th1型细胞免疫应答(12.00%,3/25),4%(1/25)受试者同时产生Th1,Th2和Th17型三种细胞免疫应答,未观察到仅产生Th2型或Th17型细胞免疫应答者。结论白念珠菌免疫优势抗原Eno诱导的T细胞免疫以Th1型和Th17型为主,提示该抗原将产生保护性细胞免疫应答,具有成为理想疫苗的潜能。Objective To observe the T cells immune response to Enolase（Eno）,an immunodominant antigen of Candida albicans.Methods Determined the frequencies of positive spot-forming cells（SFCs）of Eno antigen-specific T cells secreting IFN-γ,IL-4 and IL-17 Ain the PBMCs of 25 healthy individuals by ELISPOT assay.Results After Eno stimulation,the SFCs of IFN-γ,IL-4 and IL-17 Ain 25 healthy persons were 14.00（8.50,39.00）,0（0,0）and 2（1,4.50）,respectively.Either the SFCs of IFN-γor those of IL-17 Awere significantly higher than those of IL-4（P〈0.05）.The difference between SFCs of IFN-γand those of IL-17 A was also significant（P=0）.The response rates of IFN-γ,IL-4 and IL-17 A were 100%（25/25）,4.00%（1/25）and 88.00%（22/25）,respectively.The difference between either IFN-γor IL-17 Aand IL-4 was significant（values all P〈0.05）.Eno induced strong response（SCFs≥20）for IFN-γin 10 healthy individuals（40.00%,10/25）,but failed to induce strong response for IL-17 Aand IL-4 in all the volunteers.Major healthy individuals（84.00%,21/25）showed both Th1 and Th17 cells response against Eno,12.00%（3/25）showing Th1 cells response in isolation,and none showed Th2 or Th17 cells response individually.Conclusion Eno of Candida albicans could induce immunodominant responses of Th1 and Th17 cells,which was considered to provide protection to IC.Eno might be a potential protective vaccine against IC.

关 键 词：烯醇化酶 酶联免疫斑点技术 干扰素-γ 白细胞介素-4 白细胞介素-17 白念珠菌 

分 类 号：R379.4[医药卫生—病原生物学] R392.12[医药卫生—基础医学]