Machy P, Pierres M, Barbet J, Leserman L D
J Immunol. 1982 Nov;129(5):2098-102.
Protein A from S. aureus was covalently coupled to small sonicated liposomes containing methotrexate (MTX) and carboxyfluorescein (CF). These liposomes were incubated with mitogen-stimulated CBA T or B cells preincubated with one of a panel of murine IgG2 monoclonal antibodies directed at distinct sites on H-2Kk, I-Ak, or I-Ek molecules. Liposomes became bound to the cells as a consequence of the interaction of protein A on the liposome and antibody on the cell. The number of bound liposomes was measured by fluorescence, and the endocytosis of liposomes was evaluated by the MTX-mediated reduction in radiolabeled deoxyuridine incorporation. T cells were sensitive to MTX in liposomes bound to all anti-H-2Kk antibodies tested, but B cells were essentially insensitive, even though they expressed more H-2Kk, bound more liposomes directed at this molecule, and were equally sensitive to free MTX. By contrast, B cells were sensitive to liposome-encapsulated MTX when liposomes were bound to any antibody directed at I-Ak. The situation was more complex with respect to I-Ek. Antibodies directed at one part of the molecule were quite effective at drug delivery, while antibodies directed at other, spatially separate domains were much less effective, even though the number of liposomes bound to these various determinants was the same. The results suggest differential regulation of the internalization of H-2K by T and B cells, and are compatible with the existence of functionally distinct I-E molecules.
来自金黄色葡萄球菌的蛋白A与含有甲氨蝶呤(MTX)和羧基荧光素(CF)的小型超声处理脂质体共价偶联。将这些脂质体与经丝裂原刺激的CBA T细胞或B细胞一起孵育,这些细胞预先用一组针对H-2Kk、I-Ak或I-Ek分子上不同位点的鼠IgG2单克隆抗体之一进行孵育。由于脂质体上的蛋白A与细胞上的抗体相互作用,脂质体与细胞结合。通过荧光测量结合的脂质体数量,并通过MTX介导的放射性标记脱氧尿苷掺入减少来评估脂质体的内吞作用。T细胞对与所有测试的抗H-2Kk抗体结合的脂质体中的MTX敏感,但B细胞基本不敏感,尽管它们表达更多的H-2Kk,结合更多针对该分子的脂质体,并且对游离MTX同样敏感。相比之下,当脂质体与任何针对I-Ak的抗体结合时,B细胞对脂质体包裹的MTX敏感。关于I-Ek的情况更为复杂。针对分子一部分的抗体在药物递送方面相当有效,而针对其他空间上分开的结构域的抗体效果则要差得多,尽管与这些不同决定簇结合的脂质体数量相同。结果表明T细胞和B细胞对H-2K内化的调节存在差异,并且与功能上不同的I-E分子的存在相一致。