Sarris A H, Palade G E
J Cell Biol. 1982 Jun;93(3):591-602. doi: 10.1083/jcb.93.3.591.
A sialoglycoprotein fraction isolated from murine (DBA/2) erythrocytic ghosts (see companion article, Sarris and Palade, 1982, J. Cell. Biol. 93:583-590) was used to raise antibodies in rabbits. By immune-IgG (serum)-[125I] protein A overlays, the antibodies were found to react positively with the four sialoglycoprotein monomers (gp-2.1, gp-2.2, gp-3.1, and gp-3.2) of the original fraction, with the sialoglycoproteins detected in erythrocytic ghosts (gp-2.1 and gp-3.1), with a diffuse component (probably a macroglycolipid) trailing around gp-3.1 in SDS polyacrylamide gel electrophoretograms of solubilized ghosts, and with a minor sialoglycoprotein hidden under this trail. IgG's isolated from immune and nonimmune rabbit sera were conjugated to tetramethylrhodamine isothiocyanate and used to survey, by fluorescence microscopy, the distribution of the cognate antigens on the three different erythroid lines known to succeed each other during the life span of the mouse. In the peripheral blood of the adult, the antibodies recognized only mature erythrocytes; they did not crossreact with either platelets, monocytes, or different types of granulocytes. In the spleen of adult anemic mice, the antibodies reacted weakly with proerythroblasts and strongly with all types of erythroblasts. In enucleating erythroblasts, antigens were preferentially segregated on the cell membrane of the nascent reticulocyte. In the 10-day-old embryo, antigens were already present on the primitive nucleated erythrocytes (produced by the blood islets of the yolk sack), and in the 14-d fetus they were found on all hepatic erythroblasts and derived non-nucleated erythrocytes. A positive immunoreaction was also obtained on Friend erythroleukemic cells, before or after induction by dimethyl sulfoxide. Nonimmune serum, or nonimmune IgGs gave negative reactions in all cases. The antibodies were species-specific: they did not crossreact with either human or rat erythrocytes.
从鼠（DBA/2）红细胞膜中分离出的一种唾液糖蛋白组分（见配套文章，萨里斯和帕拉德，1982年，《细胞生物学杂志》93:583 - 590）被用于在兔体内产生抗体。通过免疫IgG（血清）-[125I]蛋白A覆盖法，发现这些抗体与原始组分的四种唾液糖蛋白单体（gp - 2.1、gp - 2.2、gp - 3.1和gp - 3.2）、在红细胞膜中检测到的唾液糖蛋白（gp - 2.1和gp - 3.1）、在溶解的红细胞膜的SDS聚丙烯酰胺凝胶电泳图谱中尾随gp - 3.1的一种弥散成分（可能是一种大糖脂）以及位于该条带下方的一种次要唾液糖蛋白发生阳性反应。从免疫和非免疫兔血清中分离出的IgG与异硫氰酸四甲基罗丹明偶联，并通过荧光显微镜观察已知在小鼠寿命期间相继出现的三种不同红细胞系上同源抗原的分布。在成年小鼠的外周血中，抗体仅识别成熟红细胞；它们与血小板、单核细胞或不同类型的粒细胞均无交叉反应。在成年贫血小鼠的脾脏中，抗体与早幼红细胞反应较弱，与所有类型的幼红细胞反应强烈。在去核的幼红细胞中，抗原优先分布在新生网织红细胞的细胞膜上。在10日龄胚胎中，抗原已存在于原始有核红细胞（由卵黄囊血岛产生）上，在14日龄胎儿中，在所有肝幼红细胞和衍生的无核红细胞上均发现有抗原。在用二甲基亚砜诱导之前或之后的弗氏红白血病细胞上也获得了阳性免疫反应。非免疫血清或非免疫IgG在所有情况下均呈阴性反应。这些抗体具有种属特异性：它们与人类或大鼠红细胞均无交叉反应。
