靶向性腫瘤相關抗原TAG-72的scFv-CD28融合基因的真核表達載體的構建△_《中國普外基礎與臨床雜志》

作者：

1.第四軍醫大學西京醫院肝膽外科（西安710032）;;
2.第四軍醫大學西京醫院消化病研究所;;
3.第四軍醫大學病理學教研室;

關鍵詞：

單鏈抗體共刺激信號 CD28分子腫瘤相關抗原

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目的構建含抗腫瘤相關抗原TAG72單鏈抗體及CD28胞內區及跨膜區融合基因的哺乳細胞表達載體，為制備消化道腫瘤靶向性嵌合錨定T細胞，并探討其對消化道腫瘤的殺傷活性。方法應用PCR法，將抗TAG72單鏈抗體(single chain variable fragment,scFv)克隆入哺乳細胞表達載體pcDNA3.0，在5′端及3′端分別引入相應酶切位點；用RTPCR法從正常人外周血T細胞克隆CD28胞內區及跨膜區的cDNA，然后將其克隆于scFv的下游，并在5′端及3′端引入相應的酶切位點。結果抗TAG-72 scFv cDNA片段為729 bp，與已知的序列相符； CD28胞內區及跨膜區的cDNA片段為240 bp，與Genebank公布的序列一致。重組的表達載體經酶切瓊脂糖電泳測序加以證實。結論完成了抗腫瘤相關抗原TAG72 scFv及CD28胞內區及跨膜區融合基因scFvCD28pcDNA3.0的構建，為制備消化道腫瘤靶向性嵌合錨定T細胞奠定了實驗基礎。

引用本文： 徐宏勇,徐立,李開宗,竇科峰,付由池,劉彥仿. 靶向性腫瘤相關抗原TAG-72的scFv-CD28融合基因的真核表達載體的構建△. 中國普外基礎與臨床雜志, 2002, 9(6): 398-401. doi: 復制

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6.	Hombach A, Wieczarkowiecz A, Marquardt T,et al. Tumorspecific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL2 secretion and can be integrated into one combined CD28/CD3 zeta signaling receptor molecule [Ｊ]. J Immunol， 2001; 167(11)∶6123.
7.	Hombach A, Sent D, Schneider C,et al. Tcell activation by recombinant receptors: CD28 costimulation is required for interleukin 2 secretion and receptormediated Tcell proliferation but does not affect receptormediated target cell lysis [Ｊ]. Cancer Res，2001; 61(5)∶1976.

1. Xu M, Real FX, Welt S, et al. Expression of TAG72 in normal colon, transitional mucosa, and colon cancer [Ｊ]. Int J Cancer, 1989; 44(6)∶985.
2. Werther JL, RiveraMacMurray S, Bruckner H, et al. Mucinassociated sialosylTn antigen expression in gastric cancer correlates with an adverse outcome [Ｊ]. Br J Cancer,1994; 69(3)∶613.
3. Kjeldsen T, Clausen H, Hirohashi S, et al. Preparation and characterization of monoclonal antibodies directed to the tumorassociated Olinked sialosyl26 alphaNacetylgalactosaminyl (sialosylTn) epitope [Ｊ]. Cancer Res,1988; 48(8)∶2214.
4. Hombach A, Heuser C, Sircar R, et al. T cell targeting of TAG72+ tumor cells by a chimeric receptor with antibodylike specificity for a carbohydrate epitope [Ｊ]. Gastroenterology, 1997; 113(4)∶1163.
5. AlvarezVallina A, Hawkins RE. Antigenspecific targening of CD28 mediated T cell costimulation using chimeric singlechain antibody variable fragmentCD28 receptors [Ｊ]. Eur J Immunol,1996； 26(10)∶2304.
6. Hombach A, Wieczarkowiecz A, Marquardt T,et al. Tumorspecific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL2 secretion and can be integrated into one combined CD28/CD3 zeta signaling receptor molecule [Ｊ]. J Immunol， 2001; 167(11)∶6123.
7. Hombach A, Sent D, Schneider C,et al. Tcell activation by recombinant receptors: CD28 costimulation is required for interleukin 2 secretion and receptormediated Tcell proliferation but does not affect receptormediated target cell lysis [Ｊ]. Cancer Res，2001; 61(5)∶1976.

《中國普外基礎與臨床雜志》

靶向性腫瘤相關抗原TAG-72的scFv-CD28融合基因的真核表達載體的構建△

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《中國普外基礎與臨床雜志》

靶向性腫瘤相關抗原TAG-72的scFv-CD28融合基因的真核表達載體的構建△

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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