CAJ | 학술논문

目的探讨不同的非人灵长类结核感染动物模型对结核特异性抗原产生免疫应答的差异性,为选择合适的动物模型做疫苗评价提供依据. 方法采用流式细胞术对恒河猴和食蟹猴CD4+T和CD8+T细胞中CD69和HLA-DR阳性细胞进行了比较,并采用PPD和ESAT-6/CFP-10多肽库对2种感染结核分枝杆菌的实验猴外周血单个核细胞( PBMCs )刺激,使用酶联免疫斑点技术( ELISPOT)对IFN-γ进行检测. 结果流式细胞术分析显示,未感染结核分枝杆菌的恒河猴中表达CD69和HLA-DR表面标记分子的CD4+T和CD8+T细胞比例明显大于食蟹猴(P<0.01). PPD刺激感染10、11个月的实验猴PBMCs、恒河猴斑点形成单位分别是食蟹猴的3、3.5倍. ESAT-6/CFP-10多肽库刺激后,恒河猴PBMCs斑点形成单位也高于食蟹猴. 结论恒河猴PBMCs对结核特异性抗原刺激诱导产生IFN-γ比食蟹猴更为敏感,适合运用于对结核疫苗免疫指标的评价.
목적 탐토불동적비인령장류결핵감염동물모형대결핵특이성항원산생면역응답적차이성,위선택합괄적동물모형주역묘평개제공의거. 방법 채용류식세포술대항하후화식해후CD4+T화CD8+T세포중CD69화HLA-DR양성세포진행료비교,병채용PPD화ESAT-6/CFP-10다태고대2충감염결핵분지간균적실험후외주혈단개핵세포( PBMCs )자격,사용매련면역반점기술( ELISPOT)대IFN-γ진행검측. 결과 류식세포술분석현시,미감염결핵분지간균적항하후중표체CD69화HLA-DR표면표기분자적CD4+T화CD8+T세포비례명현대우식해후(P<0.01). PPD자격감염10、11개월적실험후PBMCs、항하후반점형성단위분별시식해후적3、3.5배. ESAT-6/CFP-10다태고자격후,항하후PBMCs반점형성단위야고우식해후. 결론 항하후PBMCs대결핵특이성항원자격유도산생IFN-γ비식해후경위민감,괄합운용우대결핵역묘면역지표적평개.
Objective To analyze the differences of immune responses against Mycobacterium tu-berculosis antigens induced in two different nonhuman primates and to provide rationales for the selection of suitable animal models for vaccine efficacy evaluation.Methods Expression of functional surface markers including CD69 and HLA-DR, the activation markers on CD4+and CD8+T cells from in rhesus macaques and cynomolgus monkeys were measured by flow cytometry analysis.PBMCs were isolated from rhesus ma-caques and cynomolgus monkeys with Mycobacterium tuberculosis infection and stimulated with PPD and pep-tide pools ( ESAT-6/CFP-10) .Enzyme-linked imunospot ( ELISPOT) assay was performed to detect IFN-γproducing lymphocytes.Results The CD4+and CD8+T cells isolated from rhesus macaques without Myco-bacterium tuberculosis infection expressed higher levels of CD69 and HLA-DR than those from healthy cyno-molgus monkeys (P<0.01).The numbers of IFN-γspot forming cells/106 PBMCs in rhesus macaques with Mycobacterium tuberculosis infection for 10 and 11 months were respectively 3 and 3.5 times higher than that of cynomolgus monkeys upon after the stimulation of PBMCs with PPD.The levels of IFN-γproduction by the cells from rhesus macaque group were also higher than those from cynomolgus monkey group upon after the stimulation of PBMCs with ESAT-6 or CFP-10 peptide pools.Conclusion More IFN-γproducing cells were induced in rhesus macaques than that in cynimolgus monkeys after stimulation with Mycobacterium tu-berculosis antigens.Therefore, the rhesus macaques might be a better animal model for evaluating immune responses induced by Mycobacterium tuberculosis vaccines.