Nakagawa T, Nakagawa N, Delsing G A, Volkman D, Kehrl J H
Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
J Cell Biochem. 1989 Aug;40(4):417-30. doi: 10.1002/jcb.240400403.
The expression of interferon-gamma (IFN-gamma) receptors on normal human B cells and four B cell lines was studied. Recombinant human IFN-gamma was labeled with [gamma-32P]ATP using the catalytic subunit of a cAMP-dependent protein kinase. All four B cell lines, although differing in their responsiveness to IFN-gamma, were found to express high-affinity receptors (1,000-11,000 receptors/cell). Normal unactivated B lymphocytes were also found to express constitutively high-affinity receptors, approximately 1,400 receptors per cell with an estimated affinity of 295 pM. Activation of the normal B cells in vitro with the polyclonal B cell activator, Staphylococcus aureus Cowan strain I (SAC), resulted in a slight decline in receptor number and a more pronounced fall in receptor density. One of the B cell lines and unactivated normal B cells were shown to internalize labeled IFN-gamma rapidly. Chemical cross-linking of 32P-IFN-gamma to the CB B cell line and to freshly isolated B lymphocytes revealed one major cross-linked receptor-ligand complex which had an estimated molecular weight of approximately 110 kilodaltons. This complex corresponded to a 93 kD receptor cross-linked to recombinant IFN-gamma. Our data indicate that normal B lymphocytes constitutively express an approximately 93 kD IFN-gamma receptor which is similar to the receptor present on Epstein-Barr virus-transformed B cell lines.
研究了正常人B细胞和四种B细胞系上干扰素-γ(IFN-γ)受体的表达。使用环磷酸腺苷(cAMP)依赖性蛋白激酶的催化亚基,用[γ-32P]ATP标记重组人IFN-γ。发现所有四种B细胞系尽管对IFN-γ的反应性不同,但均表达高亲和力受体(1000 - 11000个受体/细胞)。还发现正常未活化的B淋巴细胞组成性地表达高亲和力受体,每细胞约1400个受体,估计亲和力为295 pM。用多克隆B细胞激活剂金黄色葡萄球菌考恩I株(SAC)在体外激活正常B细胞,导致受体数量略有下降,受体密度更明显下降。其中一种B细胞系和未活化的正常B细胞被证明能快速内化标记的IFN-γ。将32P-IFN-γ化学交联到CB B细胞系和新鲜分离的B淋巴细胞上,揭示出一种主要的交联受体-配体复合物,其估计分子量约为110千道尔顿。该复合物对应于与重组IFN-γ交联的93 kD受体。我们的数据表明,正常B淋巴细胞组成性地表达一种约93 kD的IFN-γ受体,其与爱泼斯坦-巴尔病毒转化的B细胞系上存在的受体相似。