Observation of targeting lymphocytes to therapy nude mice bearing KATO<b>Ⅲ</b> gastric cancer cell _CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 XU Hongyong ¹ , XU Li ²

1. Department of Hepatobiliary General Surgery, The 451^th Hospital of PLA, Xi’an 710054, P. R. China;
2. Institute of Digestive Disease, Xijing Hospital, The Fourth Military Medical University, Xi’an 710054, P. R. China;

Corresponding author：

XU Hongyong, Email: hongyongxu@sohu.com

Keywords：

gastric cancer; carcinoembryonic antigen; nude mice; T lymphocytes; single chain antibody fragment

DOI：

10.7507/1007-9424.201709005

Video：

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Objective To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo. Methods The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed. Results ① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05). Conclusion CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.

Citation： XU Hongyong, XU Li. Observation of targeting lymphocytes to therapy nude mice bearing KATOⅢ gastric cancer cell . CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2018, 25(3): 283-288. doi: 10.7507/1007-9424.201709005 Copy

1.	Ychou M, Duffour J, Kramar A, et al. Clinical significance and prognostic value of CA72-4 compared with CEA and CA19-9 in patients with gastric cancer. Dis Markers, 2000, 16(3-4): 105-110.
2.	Baiocchi GL, Marrelli D, Verlato G, et al. Follow-up after gastrectomy for cancer: an appraisal of the Italian research group for gastric cancer. Ann Surg Oncol, 2014, 21(6): 2005-2011.
3.	顾虎, 李正东, 姜志宝, 等. 早期肿瘤标志物反应与胃癌术后患者预后的相关性. 中国肿瘤外科杂志, 2013, 5(6): 347-351.
4.	Nam DH, Lee YK, Park JC, et al. Prognostic value of early postoperative tumor marker response in gastric cancer. Ann Surg Oncol, 2013, 20(12): 3905-3911.
5.	June CH, Bluestone JA, Nadler LM, et al. The B7 and CD28 receptor families. Immunol Today, 1994, 15(7): 321-331.
6.	徐立, 徐宏勇, 樊代明. 胃肠道肿瘤靶向抗癌胚抗原单链抗体与 T 细胞表面受体基因的融合分子构建. 中华消化杂志, 2004, 24(2): 75-77.
7.	徐宏勇, 徐立, 高建宏, 等. 抗癌胚抗原单链抗体的原核表达及对人胃癌的检测. 世界华人消化杂志, 2006, 14(18): 1780-1784.
8.	徐宏勇, 徐立, 高建宏, 等. 嵌合锚定 T 细胞致癌胚抗原阳性胃癌细胞的凋亡效应. 中华医学杂志, 2007, 87(15): 1053-1057.
9.	徐宏勇, 徐立, 李开宗. 肿瘤相关糖蛋白-72 嵌合锚定 T 细胞对肝癌的抑癌效应. 胃肠病学和肝病学杂志, 2014, 23(5): 481-485.
10.	Kucera R, Smid D, Topolcan O, et al. Searching for new biomarkers and the use of multivariate analysis in gastric cancer diagnostics. Anticancer Res, 2016, 36(4): 1967-1971.
11.	Yazaki PJ, Sherman MA, Shively JE, et al. Humanization of the anti-CEA T84.66 antibody based on crystal structure data. Protein Eng Des Sel, 2004, 17(5): 481-489.
12.	Yazaki PJ, Kassa T, Cheung CW, et al. Biodistribution and tumor imaging of an anti-CEA single-chain antibody-albumin fusion protein. Nucl Med Biol, 2008, 35(2): 151-158.
13.	Pavoni E, Flego M, Dupuis ML, et al. Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein. BMC Cancer, 2006, 6: 41.
14.	Hombach A, Heuser C, Marquardt T, et al. CD4⁺ T cells engrafted with a recombinant immunoreceptor efficiently lyse target cells in a MHC antigen- and Fas-independent fashion. J Immunol, 2001, 167(2): 1090-1096.
15.	Walther W, Stein U. Therapeutic genes for cancer gene therapy. Mol Biotechnol, 1999, 13(1): 21-28.
16.	Bethune MT, Joglekar AV. Personalized T cell-mediated cancer immunotherapy: progress and challenges. Curr Opin Biotechnol, 2017, 48: 142-152.
17.	Petersen CC, Petersen MS, Agger R, et al. Accumulation in tumor tissue of adoptively transferred T cells: A comparison between intravenous and intraperitoneal injection. J Immunother, 2006, 29(3): 241-249.
18.	Virgilio E, Proietti A, D'Urso R, et al. Measuring intragastric tumor markers in gastric cancer patients: a systematic literature review on significance and reliability. Anticancer Res, 2017, 37(6): 2817-2821.
19.	Crepeau RL, Ford ML. Challenges and opportunities in targeting the CD28/CTLA-4 pathway in transplantation and autoimmunity. Expert Opin Biol Ther, 2017, 17(8): 1001-1012.
20.	Hombach AA, Abken H. Costimulation by chimeric antigen receptors revisited the T cell antitumor response benefits from combined CD28-OX40 signalling. Int J Cancer, 2011, 129(12): 2935-2944.
21.	Kofler DM, Chmielewski M, Rappl G, et al. CD28 costimulation Impairs the efficacy of a redirected t-cell antitumor attack in the presence of regulatory T cells which can be overcome by preventing Lck activation. Mol Ther, 2011, 19(4): 760-767.
22.	Kakimi K, Karasaki T, Matsushita H, et al. Advances in personalized cancer immunotherapy. Breast Cancer, 2017, 24(1): 16-24.
23.	William-Faltaos S, Rouillard D, Lechat P, et al. Cell cycle arrest and apoptosis induced by oxaliplatin (L-OHP) on four human cancer cell lines. Anticancer Res, 2006, 26(3A): 2093-2099.
24.	Willemsen RA, Weijtens ME, Ronteltap C, et al. Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR. Gene Ther, 2000, 7(16): 1369-1377.
25.	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 Immunol, 2001, 167(11): 6123-6131.

1. Ychou M, Duffour J, Kramar A, et al. Clinical significance and prognostic value of CA72-4 compared with CEA and CA19-9 in patients with gastric cancer. Dis Markers, 2000, 16(3-4): 105-110.
2. Baiocchi GL, Marrelli D, Verlato G, et al. Follow-up after gastrectomy for cancer: an appraisal of the Italian research group for gastric cancer. Ann Surg Oncol, 2014, 21(6): 2005-2011.
3. 顾虎, 李正东, 姜志宝, 等. 早期肿瘤标志物反应与胃癌术后患者预后的相关性. 中国肿瘤外科杂志, 2013, 5(6): 347-351.
4. Nam DH, Lee YK, Park JC, et al. Prognostic value of early postoperative tumor marker response in gastric cancer. Ann Surg Oncol, 2013, 20(12): 3905-3911.
5. June CH, Bluestone JA, Nadler LM, et al. The B7 and CD28 receptor families. Immunol Today, 1994, 15(7): 321-331.
6. 徐立, 徐宏勇, 樊代明. 胃肠道肿瘤靶向抗癌胚抗原单链抗体与 T 细胞表面受体基因的融合分子构建. 中华消化杂志, 2004, 24(2): 75-77.
7. 徐宏勇, 徐立, 高建宏, 等. 抗癌胚抗原单链抗体的原核表达及对人胃癌的检测. 世界华人消化杂志, 2006, 14(18): 1780-1784.
8. 徐宏勇, 徐立, 高建宏, 等. 嵌合锚定 T 细胞致癌胚抗原阳性胃癌细胞的凋亡效应. 中华医学杂志, 2007, 87(15): 1053-1057.
9. 徐宏勇, 徐立, 李开宗. 肿瘤相关糖蛋白-72 嵌合锚定 T 细胞对肝癌的抑癌效应. 胃肠病学和肝病学杂志, 2014, 23(5): 481-485.
10. Kucera R, Smid D, Topolcan O, et al. Searching for new biomarkers and the use of multivariate analysis in gastric cancer diagnostics. Anticancer Res, 2016, 36(4): 1967-1971.
11. Yazaki PJ, Sherman MA, Shively JE, et al. Humanization of the anti-CEA T84.66 antibody based on crystal structure data. Protein Eng Des Sel, 2004, 17(5): 481-489.
12. Yazaki PJ, Kassa T, Cheung CW, et al. Biodistribution and tumor imaging of an anti-CEA single-chain antibody-albumin fusion protein. Nucl Med Biol, 2008, 35(2): 151-158.
13. Pavoni E, Flego M, Dupuis ML, et al. Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein. BMC Cancer, 2006, 6: 41.
14. Hombach A, Heuser C, Marquardt T, et al. CD4⁺ T cells engrafted with a recombinant immunoreceptor efficiently lyse target cells in a MHC antigen- and Fas-independent fashion. J Immunol, 2001, 167(2): 1090-1096.
15. Walther W, Stein U. Therapeutic genes for cancer gene therapy. Mol Biotechnol, 1999, 13(1): 21-28.
16. Bethune MT, Joglekar AV. Personalized T cell-mediated cancer immunotherapy: progress and challenges. Curr Opin Biotechnol, 2017, 48: 142-152.
17. Petersen CC, Petersen MS, Agger R, et al. Accumulation in tumor tissue of adoptively transferred T cells: A comparison between intravenous and intraperitoneal injection. J Immunother, 2006, 29(3): 241-249.
18. Virgilio E, Proietti A, D'Urso R, et al. Measuring intragastric tumor markers in gastric cancer patients: a systematic literature review on significance and reliability. Anticancer Res, 2017, 37(6): 2817-2821.
19. Crepeau RL, Ford ML. Challenges and opportunities in targeting the CD28/CTLA-4 pathway in transplantation and autoimmunity. Expert Opin Biol Ther, 2017, 17(8): 1001-1012.
20. Hombach AA, Abken H. Costimulation by chimeric antigen receptors revisited the T cell antitumor response benefits from combined CD28-OX40 signalling. Int J Cancer, 2011, 129(12): 2935-2944.
21. Kofler DM, Chmielewski M, Rappl G, et al. CD28 costimulation Impairs the efficacy of a redirected t-cell antitumor attack in the presence of regulatory T cells which can be overcome by preventing Lck activation. Mol Ther, 2011, 19(4): 760-767.
22. Kakimi K, Karasaki T, Matsushita H, et al. Advances in personalized cancer immunotherapy. Breast Cancer, 2017, 24(1): 16-24.
23. William-Faltaos S, Rouillard D, Lechat P, et al. Cell cycle arrest and apoptosis induced by oxaliplatin (L-OHP) on four human cancer cell lines. Anticancer Res, 2006, 26(3A): 2093-2099.
24. Willemsen RA, Weijtens ME, Ronteltap C, et al. Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR. Gene Ther, 2000, 7(16): 1369-1377.
25. 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 Immunol, 2001, 167(11): 6123-6131.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Observation of targeting lymphocytes to therapy nude mice bearing KATOⅢ gastric cancer cell

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