GLS和IL-12偶联重组耻垢分枝杆菌免疫效果的观察  

The effect of immunization by recombinant Mycobacterium smegmatis encoding GLS and IL-12 in mice

作　　者：杨春[1] 李娜[1] 伊正君[1] 何永林[1] 李俊明[1] 朱道银[1] 

机构地区：[1]重庆医科大学微生物学教研室,重庆400016

出　　处：《中国免疫学杂志》2007年第2期112-115,122,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(30400375)

摘　　要：目的:研究携带编码人天然颗粒溶素(GLS)和小鼠IL-12基因质粒(pZM03)偶联重组耻垢分枝杆菌(ATCC607)经鼻黏膜免疫小鼠后,小鼠体内的免疫状况。方法:BALB/c小鼠36只,随机分为生理盐水、pZM03、ATCC607、卡介苗(BCG)、重组ATCC607(即携带pZM03的ATCC607)、灭活重组ATCC607组;采用滴鼻法免疫小鼠,BCG组0、14天各1次,其它组0、4、14天各1次,第0天免疫后4周处死小鼠,用ELISA检测血清中IFN-γ、IL-12、IgG2a、lL-4的分泌水平和淋巴细胞PPD诱导的IFN-γ分泌水平以及肺泡灌洗液特异性SIgA的水平,用MTS法检测免疫小鼠脾淋巴细胞增殖情况。结果:重组ATCC607血清中IFN-γ、IL-12水平与BCG组无明显差异但明显高于其他组(P<0.05),IgG2a水平重组ATCC607组高于BCG组和其他组(P<0.05),各组间IL-4水平差异无统计学意义。重组ATCC607、BCG、灭活重组ATCC607及ATCC607组用PPD均可诱导小鼠脾淋巴细胞增殖,各组间差异无显著意义,但与生理盐水和pZM03组间差异有显著意义(P<0.05)。重组ATCC607组PPD诱导淋巴细胞IFN-γ明显高于其它组(P<0.05),与BCG组比较无显著差异。重组ATCC607等含菌组经鼻黏膜免疫可产生黏膜特异性SIgA。结论:重组ATCC607经鼻黏膜免疫小鼠后,机体特异性免疫特别是细胞免疫和黏膜免疫增强,为重组ATCC607治疗结核病的研究奠定了基础。Objective ：To study the immune effect of recombinant Mycobacterium smegmatis （ATCC607） carrying the plasmid encoding human granulysin（GLS） and murine IL-12 by intranasal immunization in mice. Methods： BALB/c mice were intranasally immunized with normal saline, pZM03, ATCC607, bacille calmette-guerin （ BCG）, recombinant ATCC607 and inactivation recombinant ATCC607 respectively. The mice were sacrificed 4 weeks after the first immunization. The levels of IL-12, IFN-γ, IgG2a, IL-4 in serum, SIgA in BALF（ bronchoalveolar lavage fluid） and IFN-γ released from spleen lymphocytes stimulated with PPD were detected with ELISA. The proliferation of spleen lymphocytes stimulated with PPD were detected with MTS. Results：The levels of IL-12, IFN-γ in serum in recombinant ATCC607 group and BCG group were higher than that of other groups（ P 〈 0. 05 ）, the difference between recombinant ATCC607 group and BCG group was not significant . The levels of IgG2a in recombinant ATCC607 group and BCG group were higher than that of other groups （ P 〈 0. 05 ）, especially in recombinant ATCC607 group（ P 〈 0. 05 ）. No change of IL-4 was found in all groups. The proliferation of spleen lymphocytes stimulated with PPD in ATCC607 group, BCG group, recombinant ATCC607 group and inactivated recombinant ATCC607 group was higher than the normal saline group and pZM03 group（ P 〈 0. 05 ）. IFN-γ released from spleen lymphocytes stimulated with PPD in recombinant ATCC607 group and BCG group was higher than that of other groups （ P 〈 0. 05） ,the difference between recombinant ATCC607 group and BCG group was not significant. The specific SIgA in BALF was detected in all groups made of bactera. Conclusion： The specific immunity, especially cell mediated immunity and mucosal immunity have been enhanced after intranasal immunization with recombinant ATCC607. The results establish the foundation for the therapy against tuberculosis with recombinant Mycobacterium smegmatis.

关 键 词：耻垢分枝杆菌 GLS IL-12 细胞免疫 鼻黏膜免疫 

分 类 号：R378.911[医药卫生—病原生物学]