Schreiber R E, Buku A, Unkeless J C
Department of Biochemistry, Mount Sinai School of Medicine, New York, NY 10029.
J Immunol. 1990 Jun 15;144(12):4735-41.
There are two distinct genes that encode murine low affinity Fc gamma RII, murine Fc gamma RII alpha, and murine Fc gamma RII beta, which are transcribed in specific cell lineages. Fc gamma RII alpha transcripts are present in macrophages, NK cells, and mesangial cells; Fc gamma RII beta transcripts are expressed in Fc gamma R-bearing B cells, T cells, and macrophages. We have devised a sandwich ELISA to quantify the expression of Fc gamma RII alpha protein. The ELISA is specific for Fc gamma RII alpha, and does not detect the closely related Fc gamma RII beta protein. Upon stimulation with IFN-gamma the Fc gamma RII beta- macrophage cell line J774a expressed a twelvefold enhanced level of Fc gamma RII alpha protein. Peritoneal macrophages synthesized varying amounts of Fc gamma RII alpha. High levels of Fc gamma RII alpha were observed in resident and thioglycollate-elicited peritoneal macrophages, but no Fc gamma RII alpha was detected in Bacillus Calmette Guérin-elicited macrophages. J774a cells stimulated with rIL-6 bound approximately twice as much anti-Fc gamma RII mAb 2.4G2 IgG as did unstimulated controls. However, the Fc gamma RII alpha-specific ELISA showed no change in the amount of Fc gamma RII alpha expressed. A probe encompassing the extracellular coding sequence of Fc gamma RII beta hybridized to two distinct transcripts that were elevated in rIL-6-stimulated J774a cells. One of these transcripts had the same mobility in electrophoresis as Fc gamma RII alpha mRNA and hybridized to an Fc gamma RII alpha-specific probe, whereas the other transcript was larger and did not hybridize to probes specific for either Fc gamma RII alpha or Fc gamma RII beta. Moreover, we confirmed, with an Fc gamma RII beta-specific probe, that J774a cells do not make Fc gamma RII beta mRNA. Thus, the larger transcript appears to encode a novel Fc gamma RII. We suggest that the increased level of binding of the anti-Fc gamma RII mAb 2.4G2 to rIL-6-induced cells represents translation of a Fc gamma R distinct from Fc gamma RII alpha or Fc gamma RII beta.
有两个不同的基因编码小鼠低亲和力FcγRII,即小鼠FcγRIIα和小鼠FcγRIIβ,它们在特定细胞谱系中被转录。FcγRIIα转录本存在于巨噬细胞、自然杀伤细胞和系膜细胞中;FcγRIIβ转录本在表达FcγR的B细胞、T细胞和巨噬细胞中表达。我们设计了一种夹心酶联免疫吸附测定法(ELISA)来定量FcγRIIα蛋白的表达。该ELISA对FcγRIIα具有特异性,不会检测到密切相关的FcγRIIβ蛋白。在用干扰素-γ刺激后,FcγRIIβ巨噬细胞系J774a表达的FcγRIIα蛋白水平提高了12倍。腹腔巨噬细胞合成了不同量的FcγRIIα。在驻留的和经巯基乙酸盐诱导的腹腔巨噬细胞中观察到高水平的FcγRIIα,但在卡介苗诱导的巨噬细胞中未检测到FcγRIIα。用重组白细胞介素-6(rIL-6)刺激的J774a细胞结合的抗FcγRII单克隆抗体2.4G2 IgG大约是未刺激对照细胞的两倍。然而,FcγRIIα特异性ELISA显示表达的FcγRIIα量没有变化。一个包含FcγRIIβ细胞外编码序列的探针与rIL-6刺激的J774a细胞中升高的两种不同转录本杂交。其中一种转录本在电泳中的迁移率与FcγRIIα mRNA相同,并与FcγRIIα特异性探针杂交,而另一种转录本更大,且不与针对FcγRIIα或FcγRIIβ的特异性探针杂交。此外,我们用FcγRIIβ特异性探针证实,J774a细胞不产生FcγRIIβ mRNA。因此,更大的转录本似乎编码一种新的FcγR。我们认为,抗FcγRII单克隆抗体2.4G2与rIL-6诱导细胞结合水平的增加代表了一种不同于FcγRIIα或FcγRIIβ的FcγR的翻译。