表达的人乳头瘤病毒-16型L_(1)蛋白质经树突状细胞递呈给淋巴细胞的研究

Dendritic cells present HPV-16-L_(1) protein expression to lymphocytes

目的研究人乳头瘤病毒(HPV)-16型L1基因和单纯疮疹病毒(HSV)-gD基因的2种重组蛋白和HPV-16型L1蛋白与树突状细胞(DC)和淋巴细胞的相互作用。方法带有重组质粒pCDNA3.0和pBV220的病毒基因在DH5α中42℃诱导表达,所获得的重组蛋白质与其抗体进行蛋白印迹。将重组HPV16-L_(1)蛋白与白细胞孵育,在TEM 1011电子显微镜下观察DC与淋巴细胞的相互作用。结果pBV220-HPV-L_(1)和pBV220-HSV-gD重组蛋白的相对分子量分别约为64000和13000。其浓度分别为0.59 mg/ml和0.46 mg/ml。此外,蛋白印迹表明针对HPV-16-L_(1)或HSV-gD制备的抗体能与表达的重组蛋白特异性结合。在电子显微镜TEM 1011视野观察到DC递呈HPV-16-L_(1)表达的蛋白质给淋巴细胞的现象。结论电子显微镜TEM 1011显示DC吞噬了颗粒,可能是病毒样颗粒(VLPs)。DC将HPV-16-L_(1)表达的蛋白可能递呈给了淋巴细胞,进行了信号传递。

Objective To determine HPV 16-L_(1) and HSV-gD recombinant proteins and the interaction of recombinant HPV 16-L_(1) proteinin dendritic cells(DCs)and lymphocytes.Methods The viral genes with recombinant plasmids pCDNA3.0 and pBV220 were induced and expressed at 42°C in DH5α,and the obtained recombinant protein and its antibody were determined by Western blotting.The recombinant HPV16-L_(1) protein was incubated with white blood cells,and the interaction between DCs and lymphocytes was observed under transmission electron microscope(TEM-1011).Results The molecule weights of pBV220-HPVL_(1) and pBV220-HSVgD recombinant proteins were about 64000 and 13000,and their concentrations were 0.59 mg/ml and 0.46 mg/ml,respectively.Western blotting indicated that the antibody prepared to against HPV-16L1 or HSV-gD could bind to the specificity of expressed recombinant protein.Moreover,in the field of TEM 1011,it was observed that DC presented the protein expressed by HPV-16-L_(1) to lymphocytes.Conclusion TEM 1011 shows DCs phagocytizeparticles,which may be virus-like particles(VLPs).DCs may present the protein expressed by HPV-16-L_(1) to lymphocytes for signal transmission.