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目的 :克隆和表达淋巴细胞相关免疫球蛋白样受体 (leuko cyte associatedimmunoglobulin likereceptor ,LAIR ) ,鉴定抗LAIR 1单克隆抗体 (mAb)与LAIR 2分子的免疫反应性。方法 :利用RT PCR ,从人外周血单个核细胞 (PBMC)及白血病细胞系Jurkat和HL 6 0中克隆了LAIRcDNA。将LAIR 1的胞膜外区基因及LAIR 2cDNA ,克隆入GST融合蛋白表达载体pGEX 4T 3中。以其转化E .coliBL2 1菌株后 ,在IPTG诱导下进行表达 ,表达产物用谷胱甘肽交联的Sepharose 4B纯化。用间接ELISA法 ,鉴定抗LAIR 1mAb对表达蛋白的免疫反应性。结果 :克隆并表达了LAIR 1和LAIR 2cDNA。同时克隆了 5种新的LAIR 1异型 ,其中从HL 6 0细胞克隆的 2种包含LAIR 1开放读框 (ORF) ,从Ju rkat细胞中克隆的 3种为LAIR 1cDNA片段。mAb鉴定表明LAIR 1与LAIR 2的免疫反应性不同。结论 :LAIR基因的转录、转录后的加工及表达 ,可能与个体和疾病状态有关。LAIR 1和LAIR 2免疫反应性的不同与其构象有关
OBJECTIVE: To clone and express leukocyte-associated immunoglobulin like receptor (LAIR) and identify the immunoreactivity of anti-LAIR 1 monoclonal antibody (mAb) with LAIR 2. Methods: LAIR cDNA was cloned from human peripheral blood mononuclear cells (PBMC) and leukemia cell lines Jurkat and HL 60 using RT-PCR. The extracellular domain of LAIR 1 and LAIR 2 cDNA were cloned into the GST fusion protein expression vector pGEX 4T 3. After being transformed into E. coli BL21 strain, the expression was induced by IPTG, and the expressed product was purified by glutathione-cross-Sepharose 4B. Indirect ELISA was used to identify the immunoreactivity of the anti-LAIR 1 mAb to the expressed protein. Results: LAIR 1 and LAIR 2 cDNAs were cloned and expressed. Five new variants of LAIR 1 were also cloned. Among them, two clones from HL 60 cells contained the LAIR 1 open reading frame (ORF) and three clones from Ju rkat cells were LAIR 1 cDNA fragments. mAb identification shows that LAIR 1 differs from LAIR 2 in its immunoreactivity. Conclusion: The transcription and post-transcriptional processing and expression of LAIR gene may be related to individual and disease status. The differences in the immunoreactivity between LAIR 1 and LAIR 2 are related to their conformation