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经ＲＴ－ＰＣＲ克隆小鼠淋巴细胞趋化因子（Ｌｙｍｐｈｏｔａｃｔｉｎ，Ｌｔｎ）的全长编码ｃＤＮＡ，将其置于ＣＭＶ启动子下游，插入Ｅ１区替代的腺病毒载体ｐＡｘ１ｃｗ。Ｌｔｎ重组腺病毒载体ｐＡｘ１ｃｗ．ＣＩｍＬｔｎ与经ＥｃｏＴ２２Ｉ酶切的Ａｄ５腺病毒ＤＮＡ－末端肽复合物共转染２９３细胞，制备Ｌｔｎ重组腺病毒，扩增到的病毒滴度为３．６×１０９ｐｆｕ／ｍｌ。用重组ＧＭ－ＣＳＦ从小鼠骨髓中扩增到的树突状细胞本身并不表达Ｌｔｎ；体外感染Ｌｔｎ重组腺病毒后４ｈ即可经ＲＴ－ＰＣＲ检测到Ｌｔｎ的表达，其２４ｈ的细胞培养上清体外对ＣＤ４＋Ｔ细胞和ＣＤ８＋细胞具有显著的趋化活性，表明所制备的Ｌｔｎ重组腺病毒能有效介导Ｌｔｎ在树突状细胞中的表达，为研究Ｌｔｎ基因修饰的树突状细胞诱导Ｔ细胞免疫应答奠定基础。
The full-length cDNA encoding mouse lymphocytic chemokine (Lymphotactin, Ltn) was cloned by RT-PCR and placed downstream of the CMV promoter and inserted into the adenoviral vector pAx1cw that was replaced by the E1 region. Ltn recombinant adenoviral vector pAx1cw. CImLtn was co-transfected with 293 cells with AdT adenovirus Ad5 adenovirus DNA-terminal peptide complex to prepare Ltn recombinant adenovirus. The titer of the amplified virus was 3.6×109 pfu/ml. Dendritic cells that have been amplified from mouse bone marrow with recombinant GM-CSF do not express Ltn by themselves; expression of Ltn can be detected by RT-PCR at 4 h after in vitro infection with recombinant adenovirus of Ltn, and the cell culture supernatant at 24 h can be obtained. The chemotaxis activity of CD4+ T cells and CD8+ cells was markedly increased in vitro, indicating that the prepared Ltn recombinant adenovirus can effectively mediate the expression of Ltn in dendritic cells to study the T cell immunity induced by Ltn gene-modified dendritic cells. Respond to lay the foundation.