Correa-Freire M C, Powers G A, Engelhard V H
J Immunol. 1984 Jan;132(1):69-75.
Conditions were optimized for surface binding by mouse lymphblastoid cell lines of liposomes containing purified human histocompatibility antigens. Antigens were reconstituted into lipid vesicles by detergent dialysis, and were incubated with the acceptor cell lines. After washing, the amount of HLA antigen on the cell surface was quantitated by double antibody radioimmunoassay. Uptake and surface expression was shown to be dependent upon vesicle phospholipid composition, vesicle:cell ratio, acceptor cell line, and stage of cell growth. These studies indicate that by using vesicles composed of 50% phosphatidylethanolamine/50% phosphatidylserine it is possible to introduce into the mouse EL4 cell surface about 70% of the amount of HLA expressed normally on the human lymphoblastoid line JY. The antigen is stably expressed on the surface for several hours and appears to be integrated into the cell membrane, as assessed by both fluorescence microscopy and susceptibility to lysis by using anti-HLA antibody and complement. The method used has the advantage over previously described procedures of not requiring the use of fusogenic proteins or agents such as polyethylene glycol or lysophospholipids, which might perturb overall membrane structure or properties.
对含有纯化人组织相容性抗原的脂质体与小鼠淋巴母细胞系进行表面结合的条件进行了优化。通过去污剂透析将抗原重组成脂质囊泡，并与受体细胞系一起孵育。洗涤后，通过双抗体放射免疫测定法定量细胞表面的HLA抗原量。摄取和表面表达显示取决于囊泡磷脂组成、囊泡与细胞比例、受体细胞系以及细胞生长阶段。这些研究表明，通过使用由50%磷脂酰乙醇胺/50%磷脂酰丝氨酸组成的囊泡，有可能将约70%正常表达于人类淋巴母细胞系JY上的HLA量引入小鼠EL4细胞表面。通过荧光显微镜以及使用抗HLA抗体和补体进行裂解敏感性评估，该抗原在表面稳定表达数小时，并且似乎整合到细胞膜中。所使用的方法相对于先前描述的程序具有优势，即不需要使用可能扰乱整体膜结构或性质的融合蛋白或试剂，如聚乙二醇或溶血磷脂。
