ObjectiveA mammalian expression vector encoding TAG72specific single chain variable region(scFv) fused to the transmembrane and intracellular domains of the signal transducing chain of CD28 was constructed, to generate for targeting of genetically modified T cells to gastrointestinal cancer, and to investigate the cytotoxicity against TAG72 positive target cells.MethodsThe transmembrane and intracellular domains of CD28 cDNA was amplified from human T lymphocytes using RTPCR to clone into a mammalin expression vecter, and CD28 fragments were ligated downstream of the antiTAG72 scFv cDNA and sequence verified.ResultsA 729base pair of antiTAG72 scFv was in accordance with sequence concerned; a 240base pair of cDNA of the transmembrane and intracellular domains of CD28 was confirmed as sequence concerning of Genebank.ConclusionWe constructed a mammalian expression vector encoding fusing gene to activate tumorassociated antigenspecific T lymphocyte, for generation of modified T lymphocytes to gastrointestinal tumors.
Citation：
XU Hongyong,XU Li,Li Kaizong,et al.. Construction of A Mammalian Expression Vector to Activate TumorAssociated AntigenSpecific T Lymphocytes. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2002, 9(6): 398-401. doi:
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Xu M, Real FX, Welt S, et al. Expression of TAG72 in normal colon, transitional mucosa, and colon cancer [Ｊ]. Int J Cancer, 1989; 44(6)∶985.
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| 2. |
Werther JL, RiveraMacMurray S, Bruckner H, et al. Mucinassociated sialosylTn antigen expression in gastric cancer correlates with an adverse outcome [Ｊ]. Br J Cancer,1994; 69(3)∶613.
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Kjeldsen T, Clausen H, Hirohashi S, et al. Preparation and characterization of monoclonal antibodies directed to the tumorassociated Olinked sialosyl26 alphaNacetylgalactosaminyl (sialosylTn) epitope [Ｊ]. Cancer Res,1988; 48(8)∶2214.
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Hombach A, Heuser C, Sircar R, et al. T cell targeting of TAG72+ tumor cells by a chimeric receptor with antibodylike specificity for a carbohydrate epitope [Ｊ]. Gastroenterology, 1997; 113(4)∶1163.
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AlvarezVallina A, Hawkins RE. Antigenspecific targening of CD28 mediated T cell costimulation using chimeric singlechain antibody variable fragmentCD28 receptors [Ｊ]. Eur J Immunol,1996； 26(10)∶2304.
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| 6. |
Hombach A, Wieczarkowiecz A, Marquardt T,et al. Tumorspecific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL2 secretion and can be integrated into one combined CD28/CD3 zeta signaling receptor molecule [Ｊ]. J Immunol， 2001; 167(11)∶6123.
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| 7. |
Hombach A, Sent D, Schneider C,et al. Tcell activation by recombinant receptors: CD28 costimulation is required for interleukin 2 secretion and receptormediated Tcell proliferation but does not affect receptormediated target cell lysis [Ｊ]. Cancer Res，2001; 61(5)∶1976.
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- 1. Xu M, Real FX, Welt S, et al. Expression of TAG72 in normal colon, transitional mucosa, and colon cancer [Ｊ]. Int J Cancer, 1989; 44(6)∶985.
- 2. Werther JL, RiveraMacMurray S, Bruckner H, et al. Mucinassociated sialosylTn 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 tumorassociated Olinked sialosyl26 alphaNacetylgalactosaminyl (sialosylTn) 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 antibodylike specificity for a carbohydrate epitope [Ｊ]. Gastroenterology, 1997; 113(4)∶1163.
- 5. AlvarezVallina A, Hawkins RE. Antigenspecific targening of CD28 mediated T cell costimulation using chimeric singlechain antibody variable fragmentCD28 receptors [Ｊ]. Eur J Immunol,1996； 26(10)∶2304.
- 6. Hombach A, Wieczarkowiecz A, Marquardt T,et al. Tumorspecific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL2 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. Tcell activation by recombinant receptors: CD28 costimulation is required for interleukin 2 secretion and receptormediated Tcell proliferation but does not affect receptormediated target cell lysis [Ｊ]. Cancer Res，2001; 61(5)∶1976.
