重组耻垢分枝杆菌经鼻粘膜免疫在小鼠体内的分布与表达

目的：探讨携带编码人天然颗粒溶素（GLS）和小鼠IL-12基因质粒（pZM03）的重组耻垢分枝杆菌经鼻粘膜免疫在小鼠体内的分布与表达。方法：将携带绿色荧光蛋白（GFP）基因质粒（pUV15）的重组耻垢分枝杆菌滴鼻BALB／c小鼠后2周，处死小鼠，观察肺、脾组织中荧光蛋白和耻垢分枝杆菌的分布；将携带GLS和IL-12质粒（pZM03）的重组耻垢分枝杆菌滴鼻免疫BALB／C小鼠3次，第1次免疫后4周，处死小鼠，用免疫组化检测肺、脾组织中GLS表达，用ELISA检测血清IL-12和肺泡灌洗液特异性SIgA的水平。结果：在BALB／c小鼠肺、脾组织中均检测到绿色荧光蛋白和耻垢分枝杆菌的分布，以及GLS的表达，并有血清IL-12水平升高和粘膜特异性SIgA的产生。结论：携带GFP的重组耻垢分枝杆菌可在肺和脾组织分布；携带GLS和IL-12的重组耻垢分枝杆菌经鼻粘膜免疫小鼠后，可引起呼吸道粘膜特异性抗菌免疫，以及GLS和IL-12的体内表达，为新型疫苗的研究奠定了基础.

The distribution and expression of recombinant M. smegmatis in mice receiving intranasal immunization

Objective: To observe the distribution and expression of recombinant Mycobacterium smegmatis carrying the plasmid encoding human granulysin(GLS) and murine IL-12 by intranasal immunization in mice.Methods: BALB/C mice were intranasally immunized with recombinant M.smegmatis carrying the plasmid encoding green fluorescent protein(GFP).The distribution of GFP and recombinant Mycobacterium smegmatis in the lungs and spleens was detected 14 days after the first immunization.BALB/C mice were intranasally immunized three times with recombinant M.smegmatis carrying the plasmid encoding GLS and IL-12.28 days after the first immunization,the expression of GLS in tissue,levels of IL-12 in serum and SIgA in BALF(bronchoalveolar lavage fluid)were detected with immunohistochemistry and ELISA respectively.Results: The distribution of GFP and recombinant Mycobacterium smegmatis in lung and spleen was detected.The expression of GLS,the increased of IL-12 in serum and the specific SIgA in BALF were found.Conclusion: The distribution of recombinant Mycobacterium smegmatis carrying the plasmid encoding of GFP in lung and spleen is detected.The specific anti-bacterium SIgA is generated.GLS and IL-12 are expressed in BALB/C mice after intranasal immunization with recombinant Mycobacterium smegmatis carrying the plasmid encoding GLS and IL-12.The results lay the foundation for new vaccine study.