Findelman F D, Shevach E M, Vitetta E S, Green I, Paul W E
J Exp Med. 1975 Jan 1;141(1):27-41. doi: 10.1084/jem.141.1.27.
We have previously demonstrated that guinea pig alloantisera directed at strain 2 and strain 13 membrane antigens block specific lymphocyte activation in immune response gene-controlled systems. In this communication we describe the partial characterization of the antigens against which these antisera are directed (the 2 and 13 antigens) and, in addition, that of the B antigen which by distribution resembles the human HL-A and mouse H-2 major histocompatibility antigens. Lymphoid cells from strain 2 and strain 13 guinea pigs were surface labeled with 125I by the lactoperoxidase technique. Nonidet P-40 extracts of these labeled cells were precipitated by sandwiches of strain 2 antistrain 13, strain 13 antistrain 2, or outbred anti-B antisera, followed by rabbit antiguinea pig immunoglobulin antisera. Precipitates were dissolved in sodium dodecyl sulfate (SDS) and electrophoresed on SDS polyacrylamide gels. Radioactive peaks representing the 2 and B-cell membrane antigens were obtained from strain 2 lymph node cells, as well as from a B-lymphoid cell population (L2C leukemia cells) and a T-lymphocyte population (STRAIN 2 PERITONEAL EXUDATE LYMPHOCYTES [PELs]). Radioactive peaks representing the 13 and B-cell membrane antigens were obtained from strain 13 lymph node cells and strain 13 PELs. All anti-B precipitates produced two peaks when electrophoresed on SDS polyacrylamide gels; one representing an antigen with a mol wt of approximately 45,000, and one representing an antigen with a mol wt of about 12,000. Both may be components of a single protein. All anti-2 and anti-13 precipitates produced a single peak when electrophoresed on SDS polyacrylamide gels. Both the 2 and 13 antigens were found by this technique to have mol wt of approximately 25,000. By molecular weight criteria, as well as by previously investigated distributional criteria, the B antigen is similar to the human LA and Four antigens, and to the mouse D and K antigens, and the 2 and 13 antigens are similar to the mouse Ia antigens.
我们先前已证明,针对2型和13型膜抗原的豚鼠同种抗血清可阻断免疫反应基因控制系统中的特异性淋巴细胞活化。在本通讯中,我们描述了这些抗血清所针对的抗原(2型和13型抗原)的部分特性,此外,还描述了B抗原的特性,其分布类似于人类HL - A和小鼠H - 2主要组织相容性抗原。通过乳过氧化物酶技术,用125I对2型和13型豚鼠的淋巴细胞进行表面标记。这些标记细胞的Nonidet P - 40提取物用2型抗13型、13型抗2型或远交系抗B抗血清的夹心沉淀,随后用兔抗豚鼠免疫球蛋白抗血清沉淀。沉淀物溶解于十二烷基硫酸钠(SDS)中,并在SDS聚丙烯酰胺凝胶上进行电泳。代表2型和B细胞膜抗原的放射性峰,从2型淋巴结细胞以及B淋巴细胞群体(L2C白血病细胞)和T淋巴细胞群体(2型腹膜渗出淋巴细胞[PELs])中获得。代表13型和B细胞膜抗原的放射性峰,从13型淋巴结细胞和13型PELs中获得。所有抗B沉淀物在SDS聚丙烯酰胺凝胶上电泳时产生两个峰;一个代表分子量约为45,000的抗原,另一个代表分子量约为12,000的抗原。两者可能是单一蛋白质的组成部分。所有抗2型和抗13型沉淀物在SDS聚丙烯酰胺凝胶上电泳时产生一个峰。通过该技术发现,2型和13型抗原的分子量均约为25,000。根据分子量标准以及先前研究的分布标准,B抗原类似于人类LA和Four抗原,以及小鼠D和K抗原,而2型和13型抗原类似于小鼠Ia抗原。