刚地弓形虫排泄分泌抗原刺激NK细胞过继转输对小鼠B16F10黑色素瘤生长的抑制作用

doi:10.12140/j.issn.1000-7423.2019.04.013

中国寄生虫学与寄生虫病杂志 ›› 2019, Vol. 37 ›› Issue (4): 444-447.doi: 10.12140/j.issn.1000-7423.2019.04.013

刚地弓形虫排泄分泌抗原刺激NK细胞过继转输对小鼠B16F10黑色素瘤生长的抑制作用

焦玉萌(), 夏惠, 王雪梅, 陶志勇, 朱琳, 方强^*()

蚌埠医学院病原生物学教研室,安徽省感染与免疫重点实验室,蚌埠 233030

收稿日期:2019-01-09 出版日期:2019-08-30 发布日期:2019-09-05
通讯作者: 方强
作者简介:
作者简介：焦玉萌（1978-）,女,硕士,副教授,从事病原生物学研究工作。E-mail: jymbbyxy@163.com
基金资助:
安徽省高校科研创新平台团队项目（No. 2016-40）;安徽省高校自然科学重点研究项目（No. KJ2015A235）;蚌埠医学院科研基金项目（No. BYKL1407ZD）;安徽省高校大学生创新训练计划项目（No. 201610367021,No. 201510367047）

Therapeutic effect of NK cells stimulated by Toxoplasma gondii excretory/secretory antigens on B16F10 melanoma in mice

Yu-meng JIAO(), Hui XIA, Xue-mei WANG, Zhi-yong TAO, Lin ZHU, Qiang FANG^*()

Department of Microbiology and Parasitology, Bengbu Medical College, Anhui Key Laboratory of Infection and Immunity, Bengbu 233030, China

Received:2019-01-09 Online:2019-08-30 Published:2019-09-05
Contact: Qiang FANG
Supported by:
Supported by the Scientifc Research Innovation Team Project of Anhui Colleges and Universities(No. 2016-40), Key Research Project of Natural Science in Colleges and Universities of Anhui Province (No. KJ2015A235), Bengbu Medical College Scientific Research Fund (No. BYKL1407ZD) and Undergraduate Students Innovation Training Program of Anhui Higher Education Institutions(No. 201610367021, No. 201510367047)

摘要/Abstract

摘要：

目的观察刚地弓形虫排泄分泌抗原（TgESA）刺激后的NK细胞对小鼠B16F10黑色素瘤生长的作用。方法取弓形虫RH株体外培养12 h后,从培养上清收集获得弓形虫ESA。取10只C57BL/6小鼠随机分为2组,每组5只,每只小鼠右腋窝皮下接种B16F10黑素瘤细胞2 × 10⁵个。接种后第7天,随机取1组小鼠腹腔注射100 μl TgESA,另1组注射等量的PBS。接种后第14天,无痛处死小鼠,无菌条件下制备小鼠脾细胞悬液,分离2组荷瘤鼠的NK细胞,分别记为NK_B16F10和NK_ESA,用于后续过继转输实验。取30只C57BL/6小鼠随机分为对照组、NK_B16F10组和NK_ESA组,每组10只。3组小鼠均右腋窝皮下接种B16F10细胞2 × 10⁵个/鼠。其中NK_B16F10组和NK_ESA组小鼠同时分别尾静脉注射2 × 10⁵个NK_B16F10和NK_ESA。观察瘤体生长情况以及各组小鼠死亡数和死亡时间,共观察35 d。结果 NK细胞过继转输后,NK_ESA组和NK_B16F10组小鼠平均出瘤时间分别为（14.70 ± 0.95）、（12.60 ± 0.70）d,均晚于对照组的（8.50 ± 0.85）d（P < 0.05）。3组小鼠出瘤后,瘤体均不断增长,至第35天,NK_B16F10组和NK_ESA组的平均瘤体面积分别为（686.53 ± 17.84）和（577.79 ± 49.70）mm²,均小于对照组的（787.84 ± 19.94）mm²（P < 0.05）。对照组、NK_B16F10组和NK_ESA组小鼠分别于接种B16F10细胞后第24、27和30天开始出现死亡,至第35天,存活的小鼠数分别为3、4和6只。结论弓形虫ESA刺激的NK细胞过继转输小鼠后可较为明显地抑制B16F10黑色素瘤瘤体的生长。

关键词: 刚地弓形虫, 排泄分泌抗原, 自然杀伤细胞, 黑色素瘤

Abstract:

Objective To observe the therapeutic effect of natural killer (NK) cells stimulated by Toxoplasma gondii excretory/secretory antigens (TgESA) on the growth of B16F10 melanoma in mice. Methods TgESA was obtained from the culture of T. gondii tachyzoites in vitro for 12 hours. The tumor-bearing animal models were established by direct injection of melanoma B16F10 cells into the right axillary of ten mice (2 × 10⁵/mouse). Five of the melanoma-inoculated mice were intraperitoneally treated with TgESA(100 μl/mouse) 7 days after inoculation of B16F10 cells, the rest 5 mice were treated with PBS only as control. One week after the treatment, all mice were euthanized, and the NK cells were isolated from the splenocytes of two groups (named NK_B16F10 and NK_ESA). Total 30 naïve C57BL/6 mice were randomly divided into 3 groups with 10 mice each group. Each mouse was inoculated with 2 × 10⁵ B16F10 cells subcutaneously into axillary. One week after melanoma inoculation, one group of mice were passive transferred with 2 × 10⁵ of NK_B16F10 cells through tail vein injection (group of NK_B16F10). Another group of mice were transferred with the same number of NK_ESA cells (group of NK_ESA). The third group of mice were given with PBS only as control. After passive transfer, the tumor growth, mortality and survival rate in the transferred mice were observed and recorded for 35 days. Results The visible time of tumor in mice of NK_ESA group [(14.70 ± 0.95) d] was significantly longer than that in NK_B16F10 group [(12.60 ± 0.70) d] and in control group [(8.50 ± 0.85) d] (P < 0.05). The tumor size in NK_ESA group [(577.79 ± 49.70) mm²] was also significantly smaller than that in NK_B16F10 group [(686.53 ± 17.84) mm²] and in control group [(787.84 ± 19.94) mm²] at D35. The delayed appearance of tumor and smaller tumor size in group of NK_B16F10 was also significant compared to control group. Mice began to die at D24 for control group, D27 for NK_B16F10 group and D30 for NK_ESA group after tumor inoculation, and the number of mice remain alive was 3, 4 and 6 respectively at terminal time D35 with significant difference between NK_ESA group and control group.Conclusion Passive transfer of NK cells stimulated by TgESA conferred therapeutic effect on B16F10 melanoma in recipient mice.

Key words: Toxoplasma gondii, Excretory/secretory antigens, Natural killer cells, Melanoma

中图分类号:

R382.5

焦玉萌, 夏惠, 王雪梅, 陶志勇, 朱琳, 方强. 刚地弓形虫排泄分泌抗原刺激NK细胞过继转输对小鼠B16F10黑色素瘤生长的抑制作用[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(4): 444-447.

Yu-meng JIAO, Hui XIA, Xue-mei WANG, Zhi-yong TAO, Lin ZHU, Qiang FANG. Therapeutic effect of NK cells stimulated by Toxoplasma gondii excretory/secretory antigens on B16F10 melanoma in mice[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2019, 37(4): 444-447.

图/表 3

参考文献 35

[1]	Kim K, Weiss LM.Toxoplasma: the next 100 years[J]. Microbes Infect, 2008, 10(9): 978-984.
[2]	Blader IJ, Koshy AA.Toxoplasma gondii development of its replicative niche: in its host cell and beyond[J]. Eukaryotic Cell, 2014, 13(8): 965-976.
[3]	Ferguson DJ.Toxoplasma gondii: 1908-2008, homage to Nicolle, Manceaux and Splendore[J]. Mem Inst Oswaldo Cruz, 2009, 104(2): 133-148.
[4]	Darani HY, Shirzad H, Mansoori F, et al. Effects of Toxoplasma gondii and Toxocara canis antigens on WEHI-164 fibrosarcoma growth in a mouse model[J]. Korean J Parasitol, 2009, 47(2): 175-177.
[5]	Pyo KH, Jung BK, Xin CF, et al. Prominent IL-12 production and tumor reduction in athymic nude mice after Toxoplasma gondii lysate antigen treatment[J]. Korean J Parasitol, 2014, 52(6): 605-612.
[6]	Thomas-Tikhonenko A, Hunter CA.Infection and cancer: the common vein[J]. Cytokine Growth Factor Rev, 2003, 14(1): 67-77.
[7]	Kim JO, Jung SS, Kim SY, et al. Inhibition of Lewis lung carcinoma growth by Toxoplasma gondii through induction of Th1 immune responses and inhibition of angiogenesis[J]. J Korean Med Sci, 2007, 22(Suppl): S38-S46.
[8]	Rankin EB, Yu DN, Jiang J, et al. An essential role of Th1 responses and interferon gamma in infection-mediated suppression of neoplastic growth[J]. Cancer Biol Ther, 2003, 2(6): 687-693.
[9]	张秀昌, 蔡念光, 孙黎, 等. 弓形虫诱导人白血病细胞K562凋亡的实验观察[J]. 中国寄生虫学与寄生虫病杂志, 2007, 25(3): 185-188.
[10]	武欣, 孙黎, 张力, 等. 弓形虫对4种肿瘤细胞增殖及凋亡的影响[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(2): 157-159.
[11]	葛璞, 李春莉, 冉亚林, 等. 刚地弓形虫培养上清抑制肺癌A549细胞系增殖[J]. 基础医学与临床, 2010, 30(2): 185-188.
[12]	罗强, 孙黎, 田青青, 等. 刚地弓形虫速殖子培养上清液对人胃癌细胞BGC-823增殖和凋亡的影响[J]. 中国寄生虫学与寄生虫病杂志, 2014, 32(2): 123-127.
[13]	孔璟, 高剑, 叶彬, 等. 刚地弓形虫对荷瘤小鼠肿瘤组织微血管生成的抑制作用[J]. 中国人兽共患病学报, 2011, 27(6): 522-525.
[14]	Daryani A, Hosseini AZ, Dalimi A.Immune responses against excreted/secreted antigens of Toxoplasma gondii tachyzoites in the murine model[J]. Vet Parasitol, 2003, 113(2): 123-134.
[15]	夏菁, 彭鸿娟. 刚地弓形虫毒力调节因子研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2015, 33(4): 297-300.
[16]	邱竞帆, 葛可, 王勇. 刚地弓形虫蛋白质组学研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2015, 33(3): 214-218.
[17]	蒲元华, 张德林. 弓形虫入侵宿主机制及免疫学研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(6): 480-485.
[18]	Carayannopoulos LN, Yokkoyama WM.Recognition of infected cells by natural killer cells[J]. Curr Opin Immunol, 2004, 16(1): 26-33.
[19]	Abou-Bacar A, Pfaff AW, Georges S, et al. Role of NK cells and gamma interferon in transplacental passage of Toxoplasma gondii in a mouse model of primary infection[J]. Infect Immun, 2004, 72(3): 1397-1401.
[20]	Combe CL, Curiel TJ, Moretto MM, et al. NK cells help to induce CD8+ T-Cell immunity against Toxoplasma gondii in the absence of CD4+ T Cells[J]. Infect Immun, 2005, 73(8): 4913-4921.
[21]	Sacks D, Sher A.Evasion of innate immunity by parasitic protozoa[J]. Nat Immunol, 2002, 3(11): 1041-1047.
[22]	Munoz M, Liesenfeld O, Heimesaat MM.Immunology of Toxoplasma gondii[J]. Immunol Rev, 2011, 240(1): 269-285.
[23]	焦玉萌, 章黎, 葛以跃, 等. 弓形虫排泄分泌抗原对B16F10黑素瘤小鼠CD4+CD25+Foxp3+T细胞和NK 细胞的影响[J]. 中国血吸虫病防治杂志, 2011, 23(3): 301-306.
[24]	焦玉萌, 方强, 夏惠, 等. 弓形虫ESA诱导小鼠产生的CD8+T细胞对早期黑色素瘤生长的影响[J]. 中国寄生虫学与寄生虫病杂志, 2013, 31(2): 95-98.
[25]	Li YX, Sun R.Tumor immunotherapy: new aspects of natural killer cells[J]. Chin J Cancer Res, 2018, 30(2): 173-196.
[26]	Grossenbacher SK, Aguilar EG, Murphy WJ.Leveraging natural killer cells for cancer immunotherapy[J]. Immunotherapy, 2017, 9(6): 487-497.
[27]	Caligiuri MA.Human natural killer cells[J]. Blood, 2008, 112(3): 461-469.
[28]	Zamai L, Ponti C, Mirandola P, et al. NK cells and cancer[J]. J Immunol, 2007, 178(7): 4011-4016.
[29]	Carlsten M, Malmberg KJ, Ljunggren HG.Natural killer cell-mediated lysis of freshly isolated human tumor cells[J]. Int J Cancer, 2009, 124(4): 757-762.
[30]	Pietra G, Manzini C, Vitale M, et al. Natural killer cells kill human melanoma cells with characteristics of cancer stem cells[J]. Int Immunol, 2009, 21(7): 793-801.
[31]	赵平森, 钟志雄, 翁锐强. NK细胞在肿瘤过继性免疫治疗中的临床应用与研究进展[J]. 中华肿瘤防治杂志, 2016, 23(7): 471-478.
[32]	徐道俊, 段清堂, 刘馨. NK及CAR-NK细胞在肿瘤过继免疫治疗中的运用现状及研究进展[J]. 中国新药杂志, 2017, 26(15): 1806-1813.
[33]	黄朝晖, 华东, 王丰, 等. 细胞因子诱导的NK细胞过继免疫治疗恶性肿瘤的初步应用[J]. 江南大学学报(自然科学版), 2005, 4(2): 209-211.
[34]	尚艺曼, 王子兵, 马一杰, 等. 肿瘤的过继性细胞免疫疗法[J]. 国际肿瘤学杂志, 2013, 40(2): 103-106.
[35]	王小溪, 何兴, 潘卫庆. 寄生虫感染与癌症[J]. 中国热带医学, 2019, 19(4): 392-396.

刚地弓形虫排泄分泌抗原刺激NK细胞过继转输对小鼠B16F10黑色素瘤生长的抑制作用

Therapeutic effect of NK cells stimulated by Toxoplasma gondii excretory/secretory antigens on B16F10 melanoma in mice

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 3

参考文献 35

相关文章 15

编辑推荐

Metrics

本文评价

[1]	徐方方, 陈权, 胡媛, 曹建平. Tsc22d3在日本血吸虫感染小鼠肝脏NK细胞中的表达及对NK杀伤功能的影响[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(2): 175-180.
[2]	景天宇, 牟汝涛, 张帆, 刘现兵, 胡雪梅, 张海霞, 李志丹. 孕期刚地弓形虫感染对滋养层细胞和蜕膜免疫细胞表面CD73表达的影响及其与不良妊娠的关系[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(2): 210-216.
[3]	章孝成, 胡媛, 彭荟, 沈玉娟, 刘华, 曹建平. 基于CRISPR/Cas9技术的刚地弓形虫bfd2缺陷虫株的构建及表型分析[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(2): 217-222.
[4]	张铃, 刘星卓, 吕芳丽. 非洲刚地弓形虫感染和弓形虫病流行概况[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(2): 241-251.
[5]	殷荷, 马磊, 党甜甜, 李佳铭, 赵志军. 刚地弓形虫Ⅰ/Ⅲ型ROP16调控TAF15对THP-1细胞影响及机制研究[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(1): 103-111.
[6]	李仕毓, 李静, 陆绍红, 郑斌. 宿主细胞自主免疫抗弓形虫的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(5): 653-658.
[7]	解晓曼, 孙航, 代莉莎, 朱文菊, 王利磊, 谢环环, 董宏杰, 张俊梅, 王琦, 周贝贝, 赵桂华, 徐超, 尹昆. 刚地弓形虫感染对小鼠脑组织转录本m⁶A甲基化修饰的影响[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(1): 27-35.
[8]	郑广福, 刘现兵, 姜昱竹, 李新雨, 胡雪梅, 张海霞. 刚地弓形虫感染孕鼠胎盘组织中中性粒细胞和IL-17与不良妊娠结局的关系[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(1): 48-54.
[9]	薛羽珊, 林萍, 程训佳, 冯萌. 慢性弓形虫感染对宿主中枢神经系统的损伤及其作用机制[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(5): 527-531.
[10]	姜文静, 孟雅莉, 赵利娜, 王春苗, 张晓磊. 刚地弓形虫棒状体蛋白18和膜表面抗原30复合核酸疫苗对小鼠的免疫保护作用[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(5): 532-538.
[11]	赵紫琪, 吕芳丽. 蒿甲醚脂质体体外抑制刚地弓形虫增殖作用的研究[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(4): 446-451.
[12]	张驰, 陈嘉婷, 辛紫萱, 杨莉莉, 杨梓瀚, 彭鸿娟. 弓形虫慢性感染小鼠脑转录组分析及与抑郁相关的犬尿氨酸通路的验证[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(3): 270-278.
[13]	叶井明, 何威, 刘慧媛, 鱼潇, 罗波, 刘美辰, 周必英. 猪囊尾蚴排泄分泌抗原TPx对仔猪树突状细胞活化的影响[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(3): 286-293.
[14]	杜鹃, 李佳, 吴迪, 余琦, 张玮, 白如念, 郭俊林, 刘庆斌, 雷琪莉, 谷传慧, 王萌, 赵浩军. 2022年北京市犬猫刚地弓形虫感染血清流行病学调查[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(3): 389-392.
[15]	李佳铭, 王艺璇, 杨宁爱, 马慧慧, 兰敏, 刘春兰, 赵志军. 刚地弓形虫ROP16蛋白对MH-S细胞极化和凋亡的影响及其相关机制[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(5): 579-586.