留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究

孟强, 张文杰, 吴伟康, 等. IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究[J]. 器官移植, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010
引用本文: 孟强, 张文杰, 吴伟康, 等. IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究[J]. 器官移植, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010
Meng Qiang, Zhang Wenjie, Wu Weikang, et al. Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models[J]. ORGAN TRANSPLANTATION, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010
Citation: Meng Qiang, Zhang Wenjie, Wu Weikang, et al. Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models[J]. ORGAN TRANSPLANTATION, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010

IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究

doi: 10.3969/j.issn.1674-7445.2023.01.010
基金项目: 

国家自然科学基金 82170667

详细信息
    作者简介:
    通讯作者:

    陶开山,博士,主任医师,教授,研究方向为异种肝移植,肝癌,Email: taokaishan0686@163.com

  • 中图分类号: R617

Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models

More Information
  • 摘要:   目的  探讨白细胞介素(IL)-18/IL-18结合蛋白(BP)介导自然杀伤(NK)-92MI细胞杀伤α-1, 3-半乳糖基转移酶(GGTA1)基因敲除(GTKO)猪内皮细胞的效应及可能机制。  方法  将NK-92MI细胞分为NK组、NK+IL-18组、NK+GTKO组、IL-18+NK+GTKO组和IL-18+IL-18BP+NK+GTKO组,采用实时定量逆转录聚合酶链反应(qRT-PCR)检测NK-92MI细胞中炎症相关基因的信使核糖核酸(mRNA)表达,乳酸脱氢酶(LDH)法检测NK-92MI细胞对GTKO猪内皮细胞的杀伤效应,脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法检测GTKO猪内皮细胞的凋亡情况,蛋白质印迹法检测具有杀伤效应的蛋白和凋亡相关蛋白的表达。  结果  与NK组、NK+IL-18组以及NK+GTKO组比较,IL-18+NK+GTKO组NK-92MI细胞中干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、IL-8、IL-3、IL-6和粒细胞-巨噬细胞集落刺激因子(GM-CSF)的mRNA表达水平均升高,差异均有统计学意义(均为P < 0.05)。与IL-18+NK+GTKO组比较,IL-18+IL-18BP+NK+GTKO组NK-92MI细胞中IFN-γ、TNF-α、IL-8、IL-3、IL-6和GM-CSF的mRNA表达水平均降低,差异均有统计学意义(均为P < 0.05)。与NK+GTKO组相比,IL-18+NK+GTKO组NK-92MI细胞中穿孔素、颗粒酶B和IFN-γ蛋白表达水平均升高,NK-92MI细胞对GTKO猪内皮细胞的杀伤率增加,GTKO猪内皮细胞凋亡率增加,GTKO猪内皮细胞中B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax)/Bcl-2和裂解半胱氨酸天冬氨酸蛋白酶(cleaved Caspase)-3/半胱氨酸天冬氨酸蛋白酶(Caspase)-3的比例均升高,差异均有统计学意义(均为P < 0.05)。与IL-18+NK+GTKO组比较,IL-18+IL-18BP+NK+GTKO组穿孔素、颗粒酶B、IFN-γ蛋白表达水平均降低,NK-92MI细胞对GTKO猪内皮细胞的杀伤率降低,GTKO猪内皮细胞凋亡率下降,GTKO猪内皮细胞中Bax/Bcl-2和cleaved Caspase-3/Caspase-3的比例均下降,差异均有统计学意义(均为P < 0.05)。  结论  IL-18BP可阻断IL-18诱导的NK-92MI细胞中炎症相关基因表达和NK-92MI细胞杀伤GTKO猪内皮细胞的效应。

     

  • 图  1  各组NK-92MI细胞中炎症相关基因的表达

    注:与NK组比较,aP < 0.05;与NK+IL-18组比较,bP < 0.05;与NK+GTKO组比较,cP < 0.05;与IL-18+NK+GTKO组比较,dP < 0.05。

    Figure  1.  Expression of inflammation-related genes of NK-92MI cells in each group

    图  2  各组NK-92MI细胞中杀伤效应蛋白的表达

    注:与NK+GTKO组比较,aP < 0.05;与IL-18+NK+GTKO组比较,bP < 0.05。

    Figure  2.  Expression of killer effector proteins of NK-92MI cells in each group

    图  3  各组NK-92MI细胞对GTKO猪内皮细胞的杀伤效应

    注:与NK+GTKO组比较,aP < 0.05;与IL-18+NK+GTKO组比较,bP < 0.05。

    Figure  3.  Cytotoxic effect of NK-92MI cells on GTKO porcine endothelial cells in each group

  • [1] 窦科峰, 张玄. 临床异种器官移植十大问题的思考[J]. 器官移植, 2022, 13(4): 411-416. DOI: 10.3969/j.issn.1674-7445.2022.04.001.

    DOU KF, ZHANG X. Reflection on 10 problems of clinical xenotransplantation[J]. Organ Transplant, 2022, 13(4): 411-416. DOI: 10.3969/j.issn.1674-7445.2022.04.001.
    [2] 中华医学会器官移植学分会. 中国肝移植术后并发症诊疗规范(2019版)[J]. 器官移植, 2021, 12(2): 129-133. DOI: 10.3969/j.issn.1674-7445.2021.02.002.

    Branch of Organ Transplantation of Chinese Medical Association. Diagnosis and treatment specification for postoperative complications after liver transplantation in China (2019 edition)[J]. Organ Transplant, 2021, 12(2): 129-133. DOI: 10.3969/j.issn.1674-7445.2021.02.002.
    [3] 张玄, 李霄, 王权成, 等. 猪-猴肝移植中的凝血调节障碍问题[J/CD]. 实用器官移植电子杂志, 2018, 6(5): 399-404. DOI: 10.3969/j.issn.2095-5332.2018.05.015.

    ZHANG X, LI X, WANG QC, et al. Coagulation dysregulation in pig-monkey liver transplantation[J/CD]. Pract J Organ Transplant (Electr Vers), 2018, 6(5): 399-404. DOI: 10.3969/j.issn.2095-5332.2018.05.015.
    [4] 淮国丽, 杜嘉祥, 潘登科. 基因编辑猪用于急性肝衰竭治疗的路径探讨[J]. 临床肝胆病杂志, 2022, 38(10): 2214-2218. DOI: 10.3969/j.issn.1001-5256.2022.10.004.

    HUAI GL, DU JX, PAN DK. The discussion on the genetically modified pigs for the treatment of acute liver failure[J]. J Clin Hepatol, 2022, 38(10): 2214-2218. DOI: 10.3969/j.issn.1001-5256.2022.10.004.
    [5] 章忠强, 梁硕洲, 司中洲, 等. 异种肝移植中的急性体液性排斥反应[J]. 临床肝胆病杂志, 2022, 38(10): 2210-2213. DOI: 10.3969/j.issn.1001-5256.2022.10.003.

    ZHANG ZQ, LIANG SZ, SI ZZ, et al. Acute humoral rejection in liver xenotransplantation[J]. J Clin Hepatol, 2022, 38(10): 2210-2213. DOI: 10.3969/j.issn.1001-5256.2022.10.003.
    [6] JI H, LI X, YUE S, et al. Pig BMSCs transfected with human TFPI combat species incompatibility and regulate the human TF pathway in vitro and in a rodent model[J]. Cell Physiol Biochem, 2015, 36(1): 233-249. DOI: 10.1159/000374067.
    [7] ZHANG X, COOPER DKC, DOU KF. Genetically-engineered pig-to-human organ transplantation: a new beginning[J]. Sci Bull, 2022, 67(18): 1827-1829. doi: 10.1016/j.scib.2022.08.026
    [8] HUANG H, LU Y, ZHOU T, et al. Innate immune cells in immune tolerance after liver transplantation[J]. Front Immunol, 2018, 9: 2401. DOI: 10.3389/fimmu.2018.02401.
    [9] HALMA J, PIERCE S, MCLENNAN R, et al. Natural killer cells in liver transplantation: can we harness the power of the immune checkpoint to promote tolerance?[J]. Clin Transl Sci, 2022, 15(5): 1091-1103. DOI: 10.1111/cts.13208.
    [10] FERNÁNDEZ-RUIZ M, SILVA JT, LÓPEZ-MEDRANO F, et al. Post-transplant monitoring of NK cell counts as a simple approach to predict the occurrence of opportunistic infection in liver transplant recipients[J]. Transpl Infect Dis, 2016, 18(4): 552-565. DOI: 10.1111/tid.12564.
    [11] 王光川, LI XC. 天然免疫细胞的获得性免疫属性及其在移植排斥中的作用[J]. 中华消化外科杂志, 2022, 21(8): 1044-1049. DOI: 10.3760/cma.j.cn115610-20220628-00376.

    WANG GC, LI XC. Features of acquired immune properties in innate immune cells and its roles in transplant rejection[J]. Chin J Dig Surg, 2022, 21(8): 1044-1049. DOI: 10.3760/cma.j.cn115610-20220628-00376.
    [12] 张玄, 王琳, 张洪涛, 等. 多基因编辑猪-猴心脏、肝脏、肾脏移植临床前研究初步报道[J]. 器官移植, 2021, 12(1): 51-56. DOI: 10.3969/j.issn.1674-7445.2021.01.008.

    ZHANG X, WANG L, ZHANG HT, et al. Preliminary report of preclinical trial of multi-genome engineering pig-to-macaque heart, liver and kidney transplantation[J]. Organ Transplant, 2021, 12(1): 51-56. DOI: 10.3969/j.issn.1674-7445.2021.01.008.
    [13] 吕丹, 金迪, 王柏山, 等. IL-18BP阻断由IL-18诱导大鼠胶原性关节炎对滑膜细胞凋亡表达的影响[J]. 中国免疫学杂志, 2017, 33(8): 1161-1163. DOI: 10.3969/j.issn.1000-484X.2017.08.008.

    LYU D, JIN D, WANG BS, et al. Effects of IL-18BP blockade on apoptosis of synovial cell in IL-18-induced mice with collagen-induced-arthritis[J]. Chin J Immunol, 2017, 33(8): 1161-1163. DOI: 10.3969/j.issn.1000-484X.2017.08.008.
    [14] 熊俊慧, 陈坚, 陈洁倩, 等. 人疱疹病毒6型感染及IL-18基因多态性与类风湿性关节炎易感性的关联[J]. 中华医院感染学杂志, 2022, 32(22): 3402-3407. DOI: 10.11816/cn.ni.2202-220542.

    XIONG JH, CHEN J, CHEN JQ, et al. Association of human herpes virus 6 infection, IL-18 gene polymorphisms with susceptibility to rheumatoid arthritis[J]. Chin J Nosocomiol, 2022, 32(22): 3402-3407. DOI: 10.11816/cn.ni.2202-220542.
    [15] 李璐, 李文强, 胡念丹, 等. 血清IL-18水平评估脓毒症患者病情严重程度的价值[J]. 中国急救复苏与灾害医学杂志, 2022, 17(2): 221-224, 229. DOI: 10.3969/j.issn.1673-6966.2022.02.023.

    LI L, LI WQ, HU ND, et al. The value of serum IL-18 level in evaluating the severity of sepsis[J]. Chin J Emerg Resusc Dis Med, 2022, 17(2): 221-224, 229. DOI: 10.3969/j.issn.1673-6966.2022.02.023.
    [16] IHIM SA, ABUBAKAR SD, ZIAN Z, et al. Interleukin-18 cytokine in immunity, inflammation, and autoimmunity: biological role in induction, regulation, and treatment[J]. Front Immunol, 2022, 13: 919973. DOI: 10.3389/fimmu.2022.919973.
    [17] PARK SY, HISHAM Y, SHIN HM, et al. Interleukin-18 binding protein in immune regulation and autoimmune diseases[J]. Biomedicines, 2022, 10(7): 1750. DOI: 10.3390/biomedicines10071750.
    [18] FAGGIONI R, CATTLEY RC, GUO J, et al. IL-18-binding protein protects against lipopolysaccharide- induced lethality and prevents the development of Fas/Fas ligand-mediated models of liver disease in mice[J]. J Immunol, 2001, 167(10): 5913-5920. DOI: 10.4049/jimmunol.167.10.5913.
    [19] BANDA NK, VONDRACEK A, KRAUS D, et al. Mechanisms of inhibition of collagen-induced arthritis by murine IL-18 binding protein[J]. J Immunol, 2003, 170(4): 2100-2105. DOI: 10.4049/jimmunol.170.4.2100.
    [20] 李家富, 许华, 王勇强. IL-18抑制剂对脓毒症并发血小板减少小鼠的保护作用[J]. 天津医科大学学报, 2021, 27(5): 487-491. https://www.cnki.com.cn/Article/CJFDTOTAL-TJYK202105010.htm

    LI JF, XU H, WANG YQ. Protective effects of IL-18 inhibitors on sepsis associated thrombocytopenia in mice[J]. J Tianjin Med Univ, 2021, 27(5): 487-491. https://www.cnki.com.cn/Article/CJFDTOTAL-TJYK202105010.htm
    [21] 秦爱玲, 梁丹. 探讨IL-18BP对卵巢癌的保护作用及可能被调控的机制[J]. 右江民族医学院学报, 2021, 43(4): 476-480. DOI: 10.3969/j.issn.1001-5817.2021.04.007.

    QIN AL, LIANG D. The protective effect of IL-18BP on ovarian cancer and the possible regulation mechanism[J]. J Youjiang Med Univ Natl, 2021, 43(4): 476-480. DOI: 10.3969/j.issn.1001-5817.2021.04.007.
    [22] LOPEZ KJ, CROSS-NAJAFI AA, FARAG K, et al. Strategies to induce natural killer cell tolerance in xenotransplantation[J]. Front Immunol, 2022, 13: 941880. DOI: 10.3389/fimmu.2022.941880.
    [23] MAEDA A, KOGATA S, TOYAMA C, et al. The innate cellular immune response in xenotransplantation[J]. Front Immunol, 2022, 13: 858604. DOI: 10.3389/fimmu.2022.858604.
    [24] 窦科峰, 纪洪辰. 异种肝移植的研究和进展[J]. 外科理论与实践, 2016, 21(3): 192-195. DOI: 10.3969/j.issn.1007-9610.2016.03.003.

    DOU KF, JI HC. Research and progress of liver xenotransplantation[J]. J Surg Concepts Pract, 2016, 21(3): 192-195. DOI: 10.3969/j.issn.1007-9610.2016.03.003.
    [25] 陶开山, 李霄, 窦科峰. 异种肝移植的历史与发展[J]. 器官移植, 2017, 8(2): 85-88. DOI: 10.3969/j.issn.1674-7445.2017.02.001.

    TAO KS, LI X, DOU KF. History and development of liver xenotransplantation[J]. Organ Transplant, 2017, 8(2): 85-88. DOI: 10.3969/j.issn.1674-7445.2017.02.001.
    [26] LEE BT, FIEL MI, SCHIANO TD. Antibody-mediated rejection of the liver allograft: an update and a clinico-pathological perspective[J]. J Hepatol, 2021, 75(5): 1203-1216. DOI: 10.1016/j.jhep.2021.07.027.
    [27] HOU X, TONG Q, WANG W, et al. Dihydromyricetin protects endothelial cells from hydrogen peroxide-induced oxidative stress damage by regulating mitochondrial pathways[J]. Life Sci, 2015, 130: 38-46. DOI: 10.1016/j.lfs.2015.03.007.
    [28] 张立坤, 邵东风, 王东海, 等. 绿原酸通过Bax/Bcl-2/Caspase-3信号通路对喉癌Hep-2细胞增殖、凋亡的影响[J]. 中国医院用药评价与分析, 2022, 22(10): 1206-1210. DOI: 10.14009/j.issn.1672-2124.2022.10.012.

    ZHANG LK, SHAO DF, WANG DH, et al. Effects of chlorogenic acid on proliferation and apoptosis of laryngeal cancer Hep-2 cells through Bax/Bcl-2/Caspase-3 signaling pathway[J]. Eval Anal Drug-use Hosp Chin, 2022, 22(10): 1206-1210. DOI: 10.14009/j.issn.1672-2124.2022.10.012.
    [29] FLORES-ROMERO H, HOHORST L, JOHN M, et al. Bcl-2-family protein tBID can act as a Bax-like effector of apoptosis[J]. EMBO J, 2022, 41(2): e108690. DOI: 10.15252/embj.2021108690.
    [30] SPITZ AZ, GAVATHIOTIS E. Physiological and pharmacological modulation of Bax[J]. Trends Pharmacol Sci, 2022, 43(3): 206-220. DOI: 10.1016/j.tips.2021.11.001.
  • 加载中
图(4)
计量
  • 文章访问数:  16
  • HTML全文浏览量:  13
  • PDF下载量:  1
  • 被引次数: 0
出版历程
  • 收稿日期:  2022-11-14
  • 网络出版日期:  2023-01-17
  • 刊出日期:  2023-01-15

目录

    /

    返回文章
    返回