源自小鼠胸腺细胞的Ia抗原的部分化学特性分析。
Partial chemical characterization of Ia antigens derived from murine thymocytes.
作者信息
Schwartz B D, Kask A M, Sharrow S O, David C S, Schwartz R H
出版信息
Proc Natl Acad Sci U S A. 1977 Mar;74(3):1195-9. doi: 10.1073/pnas.74.3.1195.
Previous chemical studies attempting to demonstrate Ia antigens on mouse thymocytes have given contradictory results. We attempted to resolve the question of whether Ia antigens exist on thymocytes (defined as thymus cells that bear a T cell marker) by isolating strain C3H thymocytes free of other contaminating cells using the fluorescence-activated cell sorter, and then chemically testing the purified populations for Ia antigens. Immunoglobulin-negative thymus cells and thymus cells selected with a rabbit antiserum to mouse brain were the two populations of thymocytes labeled with [3H]leucine after sorting. Radiolabeled proteins were solubilized with the non-ionic detergent Nonidet P-40, reacted with anti-Ia antiserum, and analyzed by electrophoresis on discontinuous sodium dodecyl sulfate/polyacrylamide gels. Ia antigens were recovered from both cell populations. These antigens were synthesized by thymocytes and were found on molecules composed of two chains of molecular weight 33,000 and 25,000, respectively, similar to Ia antigens derived from spleen cells. Assuming that all thymocytes bear similar amounts of Ia antigens, we estimated that they have approximately 1/50 the amount of Ia antigens that spleen cells do.
以往试图在小鼠胸腺细胞上证明Ia抗原的化学研究得出了相互矛盾的结果。我们试图通过使用荧光激活细胞分选仪分离不含其他污染细胞的C3H品系胸腺细胞，然后对纯化的细胞群体进行Ia抗原化学检测，来解决胸腺细胞（定义为带有T细胞标志物的胸腺细胞）上是否存在Ia抗原的问题。分选后，免疫球蛋白阴性胸腺细胞和用兔抗小鼠脑血清选择的胸腺细胞是两个用[3H]亮氨酸标记的胸腺细胞群体。用非离子去污剂Nonidet P-40溶解放射性标记的蛋白质，与抗Ia抗血清反应，并在不连续的十二烷基硫酸钠/聚丙烯酰胺凝胶上进行电泳分析。从两个细胞群体中都回收了Ia抗原。这些抗原由胸腺细胞合成，存在于分别由分子量为33,000和25,000的两条链组成的分子上，类似于源自脾细胞的Ia抗原。假设所有胸腺细胞携带的Ia抗原量相似，我们估计它们的Ia抗原量约为脾细胞的1/50。
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