Watts T H, McConnell H M
Proc Natl Acad Sci U S A. 1986 Dec;83(24):9660-4. doi: 10.1073/pnas.83.24.9660.
Membranes isolated from antigen-pulsed, antigen-presenting cells were extensively dialyzed and shown to retain the ability to present antigen to an I-Ad-restricted, antigen-specific T cell, 3DO-54.8. This ability to retain antigen on the membrane was duplicated in lysates of antigen-presenting cells as well as with pure I-A molecules in phospholipid vesicles. Measurement of the concentration of surface-associated fluorescent peptide on planar membranes prepared from antigen-pulsed phospholipid vesicles showed that about 1 peptide molecule was retained per 100 I-A molecules. We calculate that about 1000 I-A-associated peptide molecules are sufficient to stimulate the response of a single 3DO-54.8 cell. The association of fluorescent peptide with the surface depended on the presence of I-A and was blocked by unlabeled ovalbumin peptide or by a digest of hen egg lysozyme, added at the time of the pulse, but not after pulsing.
从经抗原脉冲处理的抗原呈递细胞中分离出的膜进行了广泛透析，并显示出保留了将抗原呈递给I-Ad限制的、抗原特异性T细胞3DO-54.8的能力。在抗原呈递细胞的裂解物以及磷脂囊泡中的纯I-A分子中也重现了在膜上保留抗原的这种能力。对由经抗原脉冲处理的磷脂囊泡制备的平面膜上表面相关荧光肽浓度的测量表明，每100个I-A分子大约保留1个肽分子。我们计算得出，大约1000个与I-A相关的肽分子足以刺激单个3DO-54.8细胞的反应。荧光肽与表面的结合取决于I-A的存在，并在脉冲时添加的未标记卵清蛋白肽或溶菌酶消化物的作用下被阻断，但在脉冲后则不会。
