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目的 :构建人胸腺素α1(Tα1)真核表达重组体 p BK- Tα1,并研究其转染人外周血淋巴细胞 (PBL)对后者免疫活性的影响。方法 :将已构建的 Tα1克隆重组体 p UC18- Tα1经 Eco R I、Hind 酶切 ,获得 Tα1片段 ,再将其克隆入真核细胞表达载体 p BK- CMV得到 p BK- Tα1重组体 ,p BK- Tα1以脂质体包裹的方法转染 PBL细胞 ,作 RT- PCR在 m RNA水平检测 p BK- Tα1在 PBL细胞中的表达。用 E玫瑰花环试验检测转染 p BK- Tα1后的 PBL细胞中T淋巴细胞表面 E受体的表达情况 ,并以酶联免疫吸附试验检测转染 p BK- Tα1后的 PBL细胞培养上清中的 IFN- γ和 IL- 2值。结果 :构建成功 Tα1真核细胞表达重组体 p BK- Tα1;在 m RNA水平证实 ,p BK- Tα1可在 PBL细胞中得以转录 ;初步证实 p BK- Tα1转染入 PBL细胞可促进 IFN- γ、IL- 2的表达和 T淋巴细胞表面 E受体的产生。结论 :p BK- Tα1在 PBL细胞内的表达可能会增强 PBL细胞的免疫活性
OBJECTIVE: To construct the eukaryotic expression recombinant pBK-Tα1 of human thymosin α1 (Tα1) and study the effect of its transfection on the immunological activity of human peripheral blood lymphocytes (PBL). Methods: The constructed Tα1 clone recombinant pUC18-Tα1 was digested with Eco RI and Hind to obtain the fragment of Tα1, which was then cloned into eukaryotic expression vector pBK-CMV to obtain pBK-Tα1 recombinant. PBK - Tα1 was transfected into PBL cells by liposome, and RT-PCR was used to detect the expression of p BK-Tα1 in PBL cells at m RNA level. The expression of E receptor on T lymphocytes in PBL cells transfected with pBK-Tα1 was detected by E-rosette test. The PBL cell culture supernatants after transfection with pBK-Tα1 were detected by enzyme-linked immunosorbent assay IFN- [gamma] and IL-2 values. Results: The recombinant plasmid pBK-Tα1 was successfully expressed in Tα1 eukaryotic cells; pBK-Tα1 was transcribed in PBL cells at m RNA level. Preliminary transfection of PBK-Tα1 into PBL cells could promote IFN-γ , IL-2 expression and T lymphocyte surface E receptor production. Conclusion: The expression of p BK-Tα1 in PBL cells may enhance the immunological activity of PBL cells