胸腺肽α1诱导MUC1特异性Th2型免疫应答

doi:10.13481/j.1671-587x.20180217

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (02): 299-304.doi: 10.13481/j.1671-587x.20180217

胸腺肽α1诱导MUC1特异性Th2型免疫应答

孙敏英¹, 周红月², 接晶², 谢飞², 翟瑞萍², 陈潭秀², 袁红艳², 台桂香²

1. 吉林大学基础医学院医学生物学实验中心, 吉林长春 130021;
2. 吉林大学基础医学院免疫学教研室, 吉林长春 130021

收稿日期:2017-04-25 出版日期:2018-03-28 发布日期:2018-03-30
通讯作者: 台桂香,教授,博士研究生导师(Tel:0431-85619476,E-mail:taiguixiang@163.com) E-mail:taiguixiang@163.com
作者简介:孙敏英(1987-),女,吉林省长春市人,助理工程师,医学硕士,主要从事医学生物学方面的研究。
基金资助:
吉林省科技厅科技攻关双十工程项目资助课题（20140201012YY）；吉林省长春市科技局科技项目资助课题（201622010100025）

Specific Th2 immune response of MUC1 induced by thmosin α1

SUN Minying¹, ZHOU Hongyue², JIE Jing², XIE Fei², ZHAI Ruiping², CHENTanxiu², YUAN Hongyan², TAI Guixiang²

1. Experimental Center of Medical Biology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China;
2. Department of Immunology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China

Received:2017-04-25 Online:2018-03-28 Published:2018-03-30

摘要/Abstract

摘要： 目的：采用黏蛋白与麦芽糖结合蛋白融合蛋白（MUC1-MBP）作为特异性抗原，研究胸腺肽α1（Tα1）加强诱导MUC1特异性免疫应答的类型，探讨其作为佐剂应用的可行性。方法：将C57BL/6小鼠随机分为生理盐水组（注射生理盐水）、MUC1-MBP+BCG组（注射MUC1-MBP＋BCG）和MUC1-MBP和Tα1组（注射MUC1-MBP＋Tα1），分别进行T细胞免疫活性、MUC1特异性抗体效价及亚类、Tα1联合MUC1-MBP抗肿瘤作用检测。第3次免疫后4~7 d无菌取脾脏组织，测定脾指数；淋巴细胞增殖反应测定各组小鼠的刺激指数（SI）；ELISA法测定小鼠脾细胞培养上清中干扰素γ（IFN-γ）、白细胞介素2（IL-2）、白细胞介素4（IL-4）和白细胞介素10（IL-10）的水平及小鼠血清中MUC1特异性抗体水平；第3次免疫后7 d，注射黑色素瘤细胞B16-MUC1，观察各组小鼠生存期。结果：与生理盐水组比较，MUC1-MBP+BCG组和MUC1-MBP+Tα1组小鼠脾指数与SI均升高（P<0.05或P<0.01），MUC1特异性SI明显升高（P<0.05）。与生理盐水组比较，MUC1-MBP+BCG组小鼠脾细胞培养上清中IFN-γ和IL-2水平均明显升高（P<0.05）；IL-4和IL-10水平均略微升高，但差异无统计学意义（P>0.05）。与生理盐水组比较，MUC1-MBP+Tα1组小鼠脾细胞培养上清中IL-4水平明显升高（P<0.01）；IFN-γ、IL-2和IL-10水平均略微升高，但差异无统计学意义（P>0.05）。MUC1-MBP+Tα1免疫小鼠能够产生抗MUC1特异性抗体且效价随浓度升高而升高。抗体亚型检测，与生理盐水组比较，MUC1-MBP+Tα1组IgG1水平明显升高（P<0.05或P<0.01），IgG2a水平无明显变化。肿瘤预防实验，与生理盐水组比较，MUC1-MBP＋BCG组和MUC1-MBP+Tα1组小鼠生存期未见明显差异。结论：MUC1-MBP联合Tα1更倾向于Th2型免疫应答，Tα1可以作为预防性疫苗佐剂使用，不适合治疗性疫苗使用。

关键词: Th2型免疫应答, 佐剂, 胸腺肽α1, 疫苗, 黏蛋白与麦芽糖结合蛋白融合蛋白

Abstract: Objective:To investigate the MUC1 specific immune response enhanced by thmosinα1(Tα1) using MUC1-MBP as the specific antigen,and to discuss the feasibility of MUC1-MBP as an adjuvant. Methods: The C57BL/6 mice were randomly divided into normal saline group,MUC1-MBP+BCG group and MUC1-MBP+Tα1 group.The T cellular immune activities,MUC1 special antibody and subclass,anti-tumor effect of Tα1 combined with MUC1-MBP were detected.4-7d after the 3rd immunization,the spleen indexes of the mice in various groups were measured; the lymphocyte proliferation response was used to detect the stimulate index(SI) of the mice in various groups; the levels of specific cytokines IFN-γ,IL-2,IL-4 and IL-10 in the supernatant of spleen cells of the mice were detected by ELISA; the level of serum MUC1-specific antibody was detected by ELISA.The C57BL/6 mice were inoculated with B16-MUC1 7 d after the last immunization and the survival of the mice was observed. Results: Compared with normal saline group,the spleen index and SI of the mice in MUC1-MBP+Tα1 and MUC1-MBP+BCG groups were significantly increased (P< 0.05 or P<0.01); the specific SI of MUC1 were significantly increased (P< 0.05).Compared with normal saline group,the levels of IFN-γ and IL-2 in the supernatant of spleen cells of the mice in MUC1-MBP+BCG group were obviously increased (P<0.05);the levels of IL-4 and IL-10 were slightly increased,but there were no significant differences (P>0.05); compared with normal saline group,the level of IL-4 in MUC1-MBP+Tα1 group was obviously increased (P< 0.01);the levels of IFN-γ,IL-2 and IL-10 were slightly increased,but there were no significant differences (P>0.05). The titer of MUC1 specific antibody was increased with the increase of concentration of Tα1. The antibody subtype detection results showed that compared with normal saline group,the level of IgG1 in MUC1-MBP+BCG group was significantly increased(P< 0.05 or P< 0.01),and the level of IgG2a had no obvious change.The tumor prevention experiment results showed that compared with normal saline group,the survival rates of the mice in MUC1-MBP+BCG group and MUC1-MBP+Tα1 group had no significant differences. Conclusion: MUC1-MBP combined with Tα1 prefers to Th2 immune responses in the mice.It indicates that Tα1 can be used as an adjuvanted preventive vaccines,but not suitable for therapeutic vaccines.

Key words: thmosin α1, adjuvant, mucin1-maltose brinding protein fusion protein, vaccines, Th2 immune response

中图分类号:

Q574

孙敏英, 周红月, 接晶, 谢飞, 翟瑞萍, 陈潭秀, 袁红艳, 台桂香. 胸腺肽α1诱导MUC1特异性Th2型免疫应答[J]. 吉林大学学报(医学版), 2018, 44(02): 299-304.

SUN Minying, ZHOU Hongyue, JIE Jing, XIE Fei, ZHAI Ruiping, CHENTanxiu, YUAN Hongyan, TAI Guixiang. Specific Th2 immune response of MUC1 induced by thmosin α1[J]. Journal of Jilin University Medicine Edition, 2018, 44(02): 299-304.

参考文献

[1] GoldsteinAL.From lab to bedside:emerging clinical applications of thymosin alpha-1[J].Expert OpinBiolTher,2009,9(5):593-608.
[2] Naylor PH,Quadrini K,Garaci E,et al.Immunopharmacology of thymosin alpha 1 and cytokine synergy[J].Ann NY Acad Sci,2007,1112:235-244.
[3] Romani L,Moretti S,Fallarino F,et al.Jack of all trades:thymosin α1 and its pleiotropy[J].Ann NY Acad Sci,2012,1269(1):1-6.
[4] Schulof RS,Lloyd MJ,Cleary PA,et al.A randomized trial to evaluate the immunorestorative properties of synthetic thymosin-alpha 1 in patients with lung cancer[J].J Biol Response Mod,1985,4(2):147-158.
[5] Maio M,Mackiewicz A,Testori A,et al.Large randomized study of thymosin alpha 1 interferon alfa or both in combination with dacarbazine in patients with metastatic melanoma[J].J Clin Oncol,2010,28:1780-1787.
[6] Stefanini GF,Foschi FG,Castelli E,et al.Alpha-1 thymosin and transcatheter arterial chenoembolization in hepatocellular carinoma patients-a preliminary experience[J].Hepatogastroenterology,1998,45(19):209-215.
[7] Cheng SQ,Wu MC,Chen H,et al.Anti-viral therapy using lamivudine and thymosin is helpful to prevent recurrence in hepatocellular carcinoma with coexisting active hepatitis B[J].Zhonghua Zhong Liu Za Zhi,2005,27(2):114-116.
[8] Acres B,Limacher JM.MUC1 as a target antigen for cancer immunotherapy[J].Expert Rev Vaccines,2005,4(4):493-502.
[9] Nath S,Mukherjee P.MUC1:a multifaceted oncoprotein with a key role in cancerprogression[J].Trends Mol Med,2014,20(6):332-342.
[10] Vassilaros S,Tsibanis A,Tsikkinis A,et al.Up to 15-year clinical follow-up of a pilot Phase Ⅲ immunotherapy study in stage Ⅱ breast cancer patients using oxidized mannan-MUC1[J].Immunotherapy,2013,5(11):1177-1182.
[11] Quoix E,Ramlau R,Westeel V,et al.Therapeutic vaccination with TG4010 and first-line chemotherapy in advanced non-small-cell lung cancer:a controlled phase 2B trial[J].Lancet Oncol,2011,12(12):1125-1133.
[12] Wurz GT,Kao CJ,Wolf M,et al.Tecemotide:Anantigen-specific cancer immunotherapy[J].Hum Vaccin Immunother,2014,10(11):3383-3393.
[13] Hu B,Wang J,Guo Y,et al.Pre-clinical toxicity and immunogenicity evaluation of a MUC1-MBP/BCG anti-tumor vaccine[J].Int Immunopharmacol,2016,33:108-118.
[14] Wang F,Ni W,Liu G,et al.Escherichia coli maltose-binding protein (MBP) directly induces mouseTh1activation through upregulating TLR2 and downregulating TLR4expressions[J].Immunobiology,2015,220(6):782-788.
[15] 台桂香,张凤吉,朱迅.重组人MUC1-MBP融合蛋白的抗肿瘤作用[J].中国肿瘤生物治疗杂志,2003,10(3):202-205.
[16] Fang F,Ma J,Ni W,et al.MUC1 and maltosebinding protein recombinant fusion protein combined with Bacillus Calmette-Guerin induces MUC1-specific and nonspecific anti-tumor immunity in mice[J].Mol Med Rep,2014,10(2):1056-1064.
[17] 赵小霞,马吉春,方芳,等.大肠杆菌麦芽糖结合蛋白(MBP)诱导小鼠Th1细胞的活化作用[J].中国免疫学杂志,2009,25(6):504-507.
[18] Ni W,Wang F,Liu G,et al.TLR9 played a more important role than TLR2 in the combination ofmaltose-bindingprotein and BCG-induced Th1 activation[J].Mol Immunol,2016,79:32-37.
[19] Li Q,Zhang H,Yu L,et al.Down-regulation of Notch signaling pathway reverses the Th1/Th2 imbalance in tuberculosis patients[J].Int Immunopharmacol,2018, 54:24-32.
[20] Li L,Jing YB,Liu J,et al.Study on the correlation of the effect of entecavir on Th1/Th2 cytokines level in the treatment of chronic hepatitis[J].Zhonghua Gan Zang Bing Za Zhi,2017,25(8):606-611.
[21] Kaminuma O,Saeki M,Nishimura T,et al.Differential contribution of adhesion molecules to Th1 and Th2 cell-mediated lung and bowel inflammation[J].Biol Pharm Bull,2017,40(10):1801-1805.

胸腺肽α1诱导MUC1特异性Th2型免疫应答

Specific Th2 immune response of MUC1 induced by thmosin α1

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