结核杆菌Ag85B基因疫苗与结核杆菌Ag85B和细胞因子GM-CSF共表达基因疫苗在小鼠体内的免疫效应比较

Comparative study on the immunogenicity and protective efficacy of the Mycobacterium tuberculosis Ag85B gene vaccine and the eukaryotic co-expression plasmid containing Mycobacterium tuberculosis Ag85B gene and Murine GM-CSF gene vaccine

目的比较结核杆菌Ag85B基因疫苗与结核杆菌Ag85B和细胞因子GM-CSF共表达基因疫苗在小鼠体内的免疫效应,为研制高效结核杆菌基因疫苗奠定基础。方法将雌性C57BL/6健康小鼠54只,随机分为3组,即结核杆菌Ag85B和细胞因子GM-CSF共表达基因疫苗(pIRES-Ag85B/GM-CSF)组、结核杆菌Ag85B基因疫苗(pIRES-Ag85B)组和空载体(pIRES)组,分别小鼠肌内注射疫苗,检测小鼠体内特异性体液免疫和细胞免疫应答相关指标,同时另取C57BL/6小鼠,随机分成4组,第1、2、3组免疫方法与上述各组免疫方法相对应,第4组每只小鼠背部皮下注射1×106CFU卡介苗(BCG),作为阳性对照,进行动物免疫保护性实验,比较分析两种基因疫苗在小鼠体内的免疫效应。结果结核杆菌Ag85B和细胞因子GM-CSF共表达基因疫苗在小鼠体内的免疫原性和免疫保护性明显强于结核杆菌Ag85B基因疫苗。结论结核杆菌Ag85B和细胞因子GM-CSF共表达基因疫苗能增强结核病基因疫苗的免疫原性和免疫保护性。

Comparative study on the immunogenicity and gene vaccine and the eukaryotic co-expression plasmid containing protective efficacy of the Mycobacterium tuberculosis Ag85B Mycobacterium tuberculosis Ag85B gene and murine GM-CSF gene Was carried out. In which C57BL/6 mice were divided into three groups：the eukaryotic co-expression plasmid containing Mycobacterium tuberculosis Ag85B gene and murine GM-CSF gene （pIRES-Ag85B/GM-CSF）group,Mycobacterium tuberculosis Ag85B gene vaccine（pIRES-Ag85B）group and pIRES group. Each C57BL/6 mouse was injected with different gene vaccine by intramuscular injection. Then, other C57BL/6 mice were divided into four groups, in which the first,second and thrid group was injected differente gene vaccine by intramuscular injection as the same way, and the fourth group injected with BCG. The immunogenicity and protective immunity of the two recombinant plasmids following genetic immunization were compared. All the mice were iniected with plRES-Ag85B/GM-CSF elicited higher antibody titers than that the mice were injected with plRES-Ag85B. Lymphocytes obtained from the spleen of plRES-Ag85B/GM-CSF immunized mice exhibited higher lymphocyte proliferative response, IFN-γ production and CTL activity than that for plRES-Ag85B immunized mice. The protective efficacy was also higher for plRES-Ag85B/GM-CSF immunized mice than that for pIRES-Ag85B immunized mice. However, the protective efficacy for pIRES-Ag85B/GM-CSF immunized mice was lower than that for BCG immunized mice. These results showed that the eukaryotic co-expression plasmid containing Mycobacteriurn tuberculosis Ag85B gene and murine GM-CSF gene vaccine could improve the immunogenicity and the immune protective efficacy of gene vaccine against Mycobacteriurn tuberculosis.