MOLECULAR AND BIOLOGICAL CHARACTERIZATION OF HUMAN 4-1BB AND ITS LIGAND
MOLECULAR AND BIOLOGICAL CHARACTERIZATION OF HUMAN 4-1BB AND ITS LIGAND
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DOI：
10.1002/eji.1830240943
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发表时间：
1994-09-01
影响因子：
5.4
通讯作者：
GOODWIN, RG
中科院分区：
文献类型：
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作者：
ALDERSON, MR;SMITH, CA;GOODWIN, RG
4-1BB最初被描述为由活化的鼠T细胞表达的cDNA，随后被证明编码整合膜蛋白的肿瘤坏死因子受体家族的成员。最近，我们确定并克隆了小鼠4-1BB（mu 4 -1BB-L）的配体，并证明它是一个新兴的家族成员的配体与肿瘤坏死因子的结构同源性。为了进一步表征4-1BB在免疫应答中的作用，我们着手克隆4-1BB-L的人同源物。然而，通过与鼠cDNA交叉杂交分离编码人4-1BB-L的cDNA的尝试是不成功的。因此，我们首先利用跨物种杂交来分离编码人4-1BB（hu 4 -1BB）的cDNA。然后使用由hu 4 -1BB的胞外部分偶联至人免疫球蛋白G1的Fc区（hu 4 -1BB.Fc）组成的融合蛋白，使用直接表达克隆策略从活化的CD 4（+）T细胞克隆中鉴定和克隆人4-1BB-L的基因。人4-1BB-L与其鼠对应物显示36%的氨基酸同一性，并定位于染色体19p13.3。Scatchard分析证明hu 4 -1BB.Fc与天然或重组人4-1BB-L的高亲和力结合。hu 4 -1BB的单克隆抗体和用hu 4 -1BB-L转染的细胞都在促分裂原共刺激的原代T细胞中诱导强烈的增殖反应。相比之下，当通过TCR/CD 3复合物的接合触发时，T细胞克隆上的4-1BB的连接增强活化诱导的细胞死亡。
4-1BB was originally described as a cDNA expressed by activated murine T cells and subsequently demonstrated to encode a member of the tumor necrosis factor receptor family of integral membrane proteins. Recently, we identified and cloned a murine ligand for 4-1BB (mu4-1BB-L) and demonstrated it to be a member of an emerging family of ligands with structural homology to tumor necrosis factor. To characterize further the role of 4-1BB in the immune response we undertook to clone the human homologue of 4-1BB-L. However, attempts to isolate a cDNA encoding the human 4-1BB-L by cross-hybridization with the murine cDNA were unsuccessful. Therefore we first utilized cross-species hybridization to isolate a cDNA encoding human 4-1BB (hu4-1BB). A fusion protein consisting of the extracellular portion of hu4-1BB coupled to the Fc region of human immunoglobulin G1 (hu4-1BB.Fc) was then used to identify and clone a gene for human 4-1BB-L from an activated CD4(+) T cell clone using a direct expression cloning strategy. Human 4-1BB-L shows 36% amino acid identity with its murine counterpart and maps to chromosome 19p13.3. Scatchard analysis demonstrated high-affinity binding of hu4-1BB.Fc to either native or recombinant human 4-1BB-L. Both monoclonal antibody to hu4-1BB and cells transfected with hu4-1BB-L induced a strong proliferative response in mitogen co-stimulated primary T cells. In contrast, ligation of 4-1BB on T cell clones enhanced activation-induced cell death when triggered by engagement of the TCR/CD3 complex.