Imai T, Fukudome K, Takagi S, Nagira M, Furuse M, Fukuhara N, Nishimura M, Hinuma Y, Yoshie O
Shionogi Institute for Medical Science, Osaka, Japan.
J Immunol. 1992 Nov 1;149(9):2879-86.
C33 Ag was originally identified by mAb inhibitory to syncytium formation induced by human T cell leukemia virus type 1. The Ag was shown to be a highly heterogeneous glycoprotein consisting of a 28-kDa protein and N-linked oligosaccharides ranging from 10 to 50 kDa. In the present study, cDNA clones were isolated from a human T cell cDNA expression library in Escherichia coli by using mAb C33. The identity of cDNA was verified by immunostaining and immunoprecipitation of transfected NIH3T3 cells with mAb. The cDNA contained an open reading frame of a 267-amino acid sequence which was a type III integral membrane protein of 29.6 kDa with four putative transmembrane domains and three putative N-glycosylation sites. The C33 gene was found to belong to a newly defined family of genes for membrane proteins, such as CD9, CD37, CD53, CD63, and TAPA-1, and was identical to R2, a cDNA recently isolated because of its strong up-regulation after T cell activation. Availability of mAb for C33 Ag enabled us to define its distribution in human leukocytes. C33 Ag was expressed in CD4+ T cells, CD19+ B cells, CD14+ monocytes, and CD16+ granulocytes. Its expression was low in CD8+ T cells and mostly negative in CD16+ NK cells. PHA stimulation enhanced the expression of C33 Ag in CD4+ T cells by about 5-fold and in CD8+ T cells by about 20-fold. PHA stimulation also induced the dramatic size changes in the N-linked sugars previously shown to accompany human T cell leukemia virus type 1-induced transformation of CD4+ T cells.
C33抗原最初是由一种抑制人1型T细胞白血病病毒诱导的合胞体形成的单克隆抗体鉴定出来的。该抗原被证明是一种高度异质性的糖蛋白,由一个28 kDa的蛋白质和10至50 kDa的N-连接寡糖组成。在本研究中,利用单克隆抗体C33从大肠杆菌中的人T细胞cDNA表达文库中分离出cDNA克隆。通过用单克隆抗体对转染的NIH3T3细胞进行免疫染色和免疫沉淀来验证cDNA的身份。该cDNA包含一个267个氨基酸序列的开放阅读框,它是一个29.6 kDa的III型整合膜蛋白,有四个推定的跨膜结构域和三个推定的N-糖基化位点。发现C33基因属于一个新定义的膜蛋白基因家族,如CD9、CD37、CD53、CD63和TAPA-1,并且与R2相同,R2是最近因其在T细胞激活后强烈上调而分离出的一个cDNA。针对C33抗原的单克隆抗体的可用性使我们能够确定其在人白细胞中的分布。C33抗原在CD4+ T细胞、CD19+ B细胞、CD14+单核细胞和CD16+粒细胞中表达。其在CD8+ T细胞中的表达较低,在CD16+ NK细胞中大多为阴性。PHA刺激使CD4+ T细胞中C33抗原的表达增加约5倍,在CD8+ T细胞中增加约20倍。PHA刺激还诱导了先前显示与人1型T细胞白血病病毒诱导的CD4+ T细胞转化相关的N-连接糖的显著大小变化。