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

清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制

李雅舒 贺伟峰 吕开阳

李雅舒, 贺伟峰, 吕开阳. 清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制[J]. 中华烧伤与创面修复杂志, 2024, 40(5): 415-424. DOI: 10.3760/cma.j.cn501225-20240121-00026.
引用本文: 李雅舒, 贺伟峰, 吕开阳. 清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制[J]. 中华烧伤与创面修复杂志, 2024, 40(5): 415-424. DOI: 10.3760/cma.j.cn501225-20240121-00026.
Li YS,He WF,Lyu KY.Role and mechanism of Vγ4 T cell depletion in epidermal tissue repair after ultraviolet damage to mouse skin[J].Chin J Burns Wounds,2024,40(5):415-424.DOI: 10.3760/cma.j.cn501225-20240121-00026.
Citation: Li YS,He WF,Lyu KY.Role and mechanism of Vγ4 T cell depletion in epidermal tissue repair after ultraviolet damage to mouse skin[J].Chin J Burns Wounds,2024,40(5):415-424.DOI: 10.3760/cma.j.cn501225-20240121-00026.

清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制

doi: 10.3760/cma.j.cn501225-20240121-00026
基金项目: 

国家自然科学基金青年科学基金项目 32000645

详细信息
    通讯作者:

    贺伟峰,Email:whe761211@hotmail.com

    吕开阳,Email:lvkaiyang@hotmail.com

Role and mechanism of Vγ4 T cell depletion in epidermal tissue repair after ultraviolet damage to mouse skin

Funds: 

Youth Science Fund Program of National Natural Science Foundation of China 32000645

More Information
  • 摘要:   目的  探究清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制。  方法  该研究为实验研究。将54只6~8周龄雌性C57BL/6J野生型小鼠按照随机数字表法分为Vγ4T细胞清除组和对照组(每组27只),分别腹腔注射亚美尼亚仓鼠抗小鼠Vγ4T细胞受体(TCR)单克隆抗体200 µg、等量同型对照IgG抗体。注射后1周(之后均在此时间点取小鼠),每组取3只小鼠(之后均从这2组另取小鼠),从背部皮肤组织和腋窝、腹股沟淋巴结中分别提取真皮细胞和淋巴结细胞,行流式细胞术检测真皮细胞和淋巴结细胞中Vγ4T细胞比例;每组取5只小鼠,观察背部皮肤情况,之后行苏木精-伊红(HE)染色观察皮肤组织结构并测量表皮组织厚度;每组取5只小鼠,提取表皮细胞后行流式细胞术检测表皮细胞中树突状表皮T细胞(DETC)比例。每组取3只小鼠,分别设为Vγ4T细胞清除+5次紫外线辐射(UVR)组和对照+5次UVR组,每日1次共行5次UVR,每日照射后即刻观察背部皮肤情况;每组取5只小鼠,分别设为Vγ4T细胞清除+1次UVR组和对照+1次UVR组,均行1次UVR后即刻行HE染色后测量表皮组织厚度。每组取3只小鼠,分别设为单纯Vγ4T细胞清除组、单纯对照组;再另取3只小鼠,分别设为Vγ4T细胞清除+1次UVR组和对照+1次UVR组;均同前处理后,采用实时荧光定量反转录PCR法检测表皮组织中胰岛素样生长因子Ⅰ(IGF-Ⅰ)、角质形成细胞生长因子(KGF)、Vγ5TCR、白细胞介素15(IL-15)、IL-1β、IL-23、自然杀伤细胞2族成员D(NKG2D)、组织相容性抗原60(H60)、小鼠UL16结合蛋白样转录子1(Mult1)、维甲酸早期诱导蛋白1(Rae1)的mRNA表达。  结果  注射后1周,Vγ4T细胞清除组小鼠真皮细胞、淋巴结细胞中Vγ4T细胞比例均明显低于对照组(t值分别为27.99、13.12,P<0.05);Vγ4T细胞清除组和对照组小鼠皮肤大体情况和组织结构无明显区别,表皮组织厚度相近(P>0.05);Vγ4T细胞清除组小鼠表皮细胞中DETC比例为(3.9±0.8)%,明显高于对照组的(1.6±0.5)%(t=4.84,P<0.05)。与对照+5次UVR组相比,Vγ4T细胞清除+5次UVR组小鼠进行1次UVR后皮肤鳞屑增多,照射2次出现鳞屑样痂皮,照射3~5次鳞屑样痂皮明显增多。行UVR后即刻,Vγ4T细胞清除+1次UVR组小鼠表皮组织厚度较对照+1次UVR组明显增加(t=11.50,P<0.05)。与单纯对照组相比,单纯Vγ4T细胞清除组小鼠表皮组织中Vγ5TCR的mRNA表达明显上调(t=41.16,P<0.05),IL-23的mRNA表达明显下调(t=6.52,P<0.05);与单纯对照组相比,对照+1次UVR组小鼠表皮组织中Vγ5TCR、KGF的mRNA表达均明显上调(t值分别为15.22、13.22,P<0.05),IGF-Ⅰ、IL-23的mRNA表达均明显下调(t值分别为3.71、4.95,P<0.05);与单纯Vγ4T细胞清除组相比,Vγ4T细胞清除+1次UVR组小鼠表皮组织中IGF-Ⅰ、KGF的mRNA表达均明显上调(t值分别为11.40、18.88,P<0.05),IL-1β的mRNA表达明显下调(t=4.42,P<0.05);与对照+1次UVR组相比,Vγ4T细胞清除+1次UVR组小鼠表皮组织中Vγ5TCR、IGF-Ⅰ、KGF的mRNA表达均明显上调(t值分别为4.52、15.24、9.43,P<0.05);4组小鼠表皮组织中IL-15的mRNA表达总体相近(P>0.05)。与单纯对照组相比,单纯Vγ4T细胞清除组表皮组织中NKG2D、Rae1的mRNA表达均明显上调(t值分别为3.67、47.40,P<0.05),对照+1次UVR组小鼠表皮组织中NKG2D、Mult1、Rae1的mRNA表达均明显上调(t值分别为5.30、6.50、9.16,P<0.05);与单纯Vγ4T细胞清除组相比,Vγ4T细胞清除+1次UVR组小鼠表皮组织中NKG2D、H60、Mult1、Rae1的mRNA表达均明显下调(t值分别为4.57、4.13、4.67、27.36,P<0.05);与对照+1次UVR组相比,Vγ4T细胞清除+1次UVR组小鼠表皮组织中NKG2D、H60、Mult1、Rae1的mRNA表达均明显下调(t值分别为5.77、8.18、12.90、8.08,P<0.05)。  结论  清除Vγ4T细胞有利于DETC增殖和毒性下调,可能促进UVR后小鼠表皮损伤修复

     

  • 参考文献(31)

    [1] WittlichM, WesterhausenS, StrehlB, et al. The GENESIS-UV study on ultraviolet radiation exposure levels in 250 occupations to foster epidemiological and legislative efforts to combat nonmelanoma skin cancer[J]. Br J Dermatol, 2023,188(3):350-360. DOI: 10.1093/bjd/ljac093.
    [2] Castejón-GriñánM, CerdidoS, Sánchez-BeltránJ, et al. Melanoma-associated melanocortin 1 receptor variants confer redox signaling-dependent protection against oxidative DNA damage[J]. Redox Biol, 2024,72: 103135. DOI: 10.1016/j.redox.2024.103135.
    [3] SlominskiRM,ChenJY,RamanC,et al.Photo-neuro-immuno-endocrinology: how the ultraviolet radiation regulates the body, brain, and immune system[J].Proc Natl Acad Sci U S A,2024,121(14):e2308374121.DOI: 10.1073/pnas.2308374121.
    [4] FrascoliM, FerrajE, MiuB, et al. Skin γδ T cell inflammatory responses are hardwired in the thymus by oxysterol sensing via GPR183 and calibrated by dietary cholesterol[J]. Immunity, 2023,56(3):562-575.e6. DOI: 10.1016/j.immuni.2023.01.025.
    [5] WeiYX,SunGY,YangY,et al.Double-negative T cells ameliorate psoriasis by selectively inhibiting IL-17A-producing γδlow T cells[J].J Transl Med,2024,22(1):328.DOI: 10.1186/s12967-024-05132-8.
    [6] PeslierH,ReichartJ,BoursotC,et al.Extensive cutaneous-mucosal and muscular involvement of gamma/delta cutaneous T-cell lymphoma on 18F-FDG PET/CT[J].Clin Nucl Med,2024,49(5):e206-e207.DOI: 10.1097/RLU.0000000000005135.
    [7] YangYL, ZhouC, ChenQ, et al. YAP1/Piezo1 involve in the dynamic changes of lymphatic vessels in UVR-induced photoaging progress to squamous cell carcinoma[J]. J Transl Med, 2023,21(1):820. DOI: 10.1186/s12967-023-04458-z.
    [8] HarmonC, ZaborowskiA, MooreH, et al. γδ T cell dichotomy with opposing cytotoxic and wound healing functions in human solid tumors[J]. Nat Cancer, 2023,4(8):1122-1137. DOI: 10.1038/s43018-023-00589-w.
    [9] PetrovićJ,SilvaJR,BannermanCA,et al.γδ T cells modulate myeloid cell recruitment but not pain during peripheral inflammation[J].Front Immunol,2019,10:473.DOI: 10.3389/fimmu.2019.00473.
    [10] SonomotoK,SongR,ErikssonD,et al.High-fat-diet‐associated intestinal microbiota exacerbates psoriasis-like inflammation by enhancing systemic γδ T cell IL-17 production[J].Cell Rep,2023,42(7):112713.DOI: 10.1016/j.celrep.2023.112713.
    [11] LiuM, LiuZH, ChenYX, et al. Dendritic epidermal T cells secreting exosomes promote the proliferation of epidermal stem cells to enhance wound re-epithelialization[J]. Stem Cell Res Ther, 2022,13(1):121. DOI: 10.1186/s13287-022-02783-6.
    [12] ChenC, MengZY, RenH, et al. The molecular mechanisms supporting the homeostasis and activation of dendritic epidermal T cell and its role in promoting wound healing[J/OL]. Burns Trauma, 2021,9:tkab009[2024-01-21]. https://pubmed.ncbi.nlm.nih.gov/34212060/.DOI: 10.1093/burnst/tkab009.
    [13] ThelenF, WitherdenDA. Get in touch with dendritic epithelial T cells![J]. Front Immunol, 2020,11:1656. DOI: 10.3389/fimmu.2020.01656.
    [14] HeiligJS, TonegawaS. Diversity of murine gamma genes and expression in fetal and adult T lymphocytes[J]. Nature, 1986,322(6082):836-840. DOI: 10.1038/322836a0.
    [15] LiYS, WuJ, LuoGX, et al. Functions of Vγ4 T cells and dendritic epidermal T cells on skin wound healing[J]. Front Immunol, 2018,9:1099. DOI: 10.3389/fimmu.2018.01099.
    [16] 王珏, 张小容, 贺伟峰, 等. 树突状表皮T细胞在创面愈合中作用机制的研究进展[J].中华烧伤杂志,2021,37(3):296-300. DOI: 10.3760/cma.j.cn501120-20200226-00092.
    [17] LiYS, WangYP, ZhouLN, et al. Vγ4 T cells inhibit the pro-healing functions of dendritic epidermal T cells to delay skin wound closure through IL-17A[J]. Front Immunol, 2018,9:240. DOI: 10.3389/fimmu.2018.00240.
    [18] YangBW, LinYM, HuangYB, et al. Extracellular vesicles modulate key signalling pathways in refractory wound healing[J/OL]. Burns Trauma, 2023,11:tkad039[2024-01-21].https://pubmed.ncbi.nlm.nih.gov/38026441/.DOI: 10.1093/burnst/tkad039.
    [19] LiuZY,LiangGP,GuiL,et al.Weakened IL-15 production and impaired mTOR Activation alter dendritic epidermal T cell homeostasis in diabetic mice[J].Sci Rep,2017,7(1):6028.DOI: 10.1038/s41598-017-05950-5.
    [20] Dhillon-LaBrooyA, BrabandKL, TantawyE, et al. Inhibition of mitochondrial translation ameliorates imiquimod-induced psoriasis-like skin inflammation by targeting Vγ4+ γδ T cells[J]. J Invest Dermatol, 2024,144(4):844-854.e2. DOI: 10.1016/j.jid.2023.09.275.
    [21] LiYS, WangJ, WangYP, et al. IL-1β/NF-κB signaling inhibits IGF-1 production via let-7f-5p in dendritic epidermal T cells[J]. J Leukoc Biol, 2022,112(6):1677-1690. DOI: 10.1002/JLB.3MA0322-171R.
    [22] LiYS, HuangZG, YanRS, et al. Vγ4 γδ T cells provide an early source of IL-17A and accelerate skin graft rejection[J]. J Invest Dermatol, 2017,137(12):2513-2522. DOI: 10.1016/j.jid.2017.03.043.
    [23] WeiXR, LiMX, ZhengZJ, et al. Senescence in chronic wounds and potential targeted therapies[J/OL]. Burns Trauma, 2022,10:tkab045[2024-01-21]. https://pubmed.ncbi.nlm.nih.gov/35187179/.DOI: 10.1093/burnst/tkab045.
    [24] NitaharaA, ShimuraH, ItoA, et al. NKG2D ligation without T cell receptor engagement triggers both cytotoxicity and cytokine production in dendritic epidermal T cells[J]. J Invest Dermatol, 2006,126(5):1052-1058. DOI: 10.1038/sj.jid.5700112.
    [25] CunninghamTJ,TabacchiM,ElianeJP,et al.Randomized trial of calcipotriol combined with 5-fluorouracil for skin cancer precursor immunotherapy[J].J Clin Invest,2017,127(1):106-116.DOI: 10.1172/JCI89820.
    [26] PeterleL,SanfilippoS,BorgiaF,et al.Alopecia areata: a review of the role of oxidative stress, possible biomarkers, and potential novel therapeutic approaches[J].Antioxidants (Basel),2023,12(1):135.DOI: 10.3390/antiox12010135.
    [27] XiangJ, QiuMH, ZhangHY. Role of dendritic epidermal T cells in cutaneous carcinoma[J]. Front Immunol, 2020,11:1266. DOI: 10.3389/fimmu.2020.01266.
    [28] IbusukiA,KawaiK,YoshidaS,et al.NKG2D triggers cytotoxicity in murine epidermal γδ T cells via PI3K-dependent, Syk/ZAP70-independent signaling pathway[J].J Invest Dermatol,2014,134(2):396-404.DOI: 10.1038/jid.2013.353.
    [29] WangYP, BaiY, LiYS, et al. IL-15 enhances activation and IGF-1 production of dendritic epidermal T cells to promote wound healing in diabetic mice[J]. Front Immunol, 2017,8:1557. DOI: 10.3389/fimmu.2017.01557.
    [30] Muñoz-RuizM, LlorianM, D'AntuonoR, et al. IFN-γ-dependent interactions between tissue-intrinsic γδ T cells and tissue-infiltrating CD8 T cells limit allergic contact dermatitis[J]. J Allergy Clin Immunol, 2023,152(6):1520-1540. DOI: 10.1016/j.jaci.2023.07.015.
    [31] NielsenMM, Dyring-AndersenB, SchmidtJD, et al. NKG2D-dependent activation of dendritic epidermal T cells in contact hypersensitivity[J]. J Invest Dermatol, 2015,135(5):1311-1319. DOI: 10.1038/jid.2015.23.
  • 1  对照组和Vγ4T细胞清除组小鼠注射后1周皮肤大体和组织病理学情况。1A、1B.分别为对照组和Vγ4T细胞清除组皮肤大体情况,两者无明显区别;1C、1D.分别为对照组和Vγ4T细胞清除组皮肤组织病理学情况,两者表皮组织厚度、组织结构无明显区别 苏木精-伊红×40

    注:对Vγ4T细胞清除组、对照组小鼠分别予以腹腔内注射亚美尼亚仓鼠抗小鼠Vγ4T细胞受体单克隆抗体、同型对照IgG抗体

    2  对照+5次UVR组和Vγ4T细胞清除+5次UVR组小鼠背部皮肤大体情况。2A、2B、2C、2D、2E.分别为对照+5次UVR组1、2、3、4、5次行UVR后即刻的皮肤大体情况,皮肤鳞屑逐渐增多;2F、2G、2H、2I、2J.分别为Vγ4T细胞清除+5次UVR组行1、2、3、4、5次UVR后即刻的皮肤大体情况,皮肤鳞屑分别较图2A、2B、2C、2D、2E明显增多

    注:对Vγ4T细胞清除+5次紫外线辐射(UVR)组、对照+5次UVR组小鼠分别予以腹腔内注射亚美尼亚仓鼠抗小鼠Vγ4T细胞受体单克隆抗体、同型对照IgG抗体1周后,均另行5次UVR

    3  对照+1次UVR组和Vγ4T细胞清除+1次UVR组小鼠行UVR后即刻皮肤组织病理学情况 苏木精-伊红×40。3A、3B.分别为对照+1次UVR组和Vγ4T细胞清除+1次UVR组皮肤组织病理学情况,图3B皮肤表皮组织厚度较图3A明显增加

    注:对Vγ4T细胞清除+1次紫外线辐射(UVR)组、对照+1次UVR组小鼠分别予以腹腔内注射亚美尼亚仓鼠抗小鼠Vγ4T细胞受体单克隆抗体、同型对照IgG抗体1周后,均另行1次UVR

    表1  4组小鼠表皮组织中Vγ5TCR和5种细胞因子的mRNA表达比较(x¯±s

    表1.   Comparison of the mRNA expressions of Vγ5 TCR and five cytokines in the epidermal tissue of mice in four groups

    组别样本数Vγ5TCRIGF-ⅠKGFIL-15IL-1βIL-23
    单纯对照组31.01±0.161.00±0.051.01±0.111.01±0.151.02±0.031.00±0.09
    单纯Vγ4T细胞清除组363.09±2.131.14±0.611.00±0.061.01±0.261.49±0.260.41±0.09
    对照+1次UVR组349.89±4.540.64±0.133.52±0.240.82±0.170.89±0.150.68±0.03
    Vγ4T细胞清除+1次UVR组366.10±2.279.03±0.776.81±0.430.83±0.130.68±0.030.57±0.07
    t141.160.330.040.316.52
    P1<0.0010.7610.9690.7720.003
    t215.223.7113.221.184.95
    P2<0.0010.021<0.0010.3040.008
    t31.3711.4018.884.421.91
    P30.242<0.001<0.0010.0120.129
    t44.5215.249.431.861.89
    P40.011<0.001<0.0010.1370.131
    注:Vγ5TCR为Vγ5T细胞受体,UVR为紫外线辐射,IGF-Ⅰ为胰岛素样生长因子Ⅰ,KGF为角质形成细胞生长因子,IL为白细胞介素;对单纯Vγ4T细胞清除组、单纯对照组小鼠分别予以腹腔内注射亚美尼亚仓鼠抗小鼠Vγ4TCR单克隆抗体、同型对照IgG抗体,并于注射后1周进行检测,对对照+1次UVR组和Vγ4T细胞清除+1次UVR组小鼠分别同单纯对照组、单纯Vγ4T细胞清除组注射后1周均另行1次UVR,然后进行检测;Vγ5TCRIGF-Ⅰ、KGF、IL-15、IL-1β、IL-23的mRNA表达的Vγ4T细胞清除主效应,F值分别为153.80、59.45、457.60、1.56、16.74、1.89,P值分别为<0.001、0.002、<0.001、0.280、0.015、0.241;UVR主效应,F值分别为479.40、2 892.00、98.50、<0.01、2.10、60.42,P值分别为<0.001、<0.001、<0.001、0.989、0.221、0.002;两者交互作用,F值分别为120.10、71.49、72.04、<0.01、9.50、16.15,P值分别为<0.001、0.001、0.001、0.976、0.037、0.016;t1值、P1值为单纯对照组与单纯Vγ4T细胞清除组比较所得;t2值、P2值为单纯对照组与对照+1次UVR组比较所得;t3值、P3值为单纯Vγ4T细胞清除组与Vγ4T细胞清除+1次UVR组比较所得,t4值、P4值为对照+1次UVR组与Vγ4T细胞清除+1次UVR组比较所得;“—”表示无此统计量值
    下载: 导出CSV

    表2  4组小鼠表皮组织中NKG2D及其3个配体的mRNA表达比较(x¯±s

    表2.   Comparison of the mRNA expressions of NKG2D and the three ligands in the epidermal tissue of mice in 4 groups

    组别样本数NKG2DH60Mult1Rae1
    单纯对照组31.03±0.251.04±0.291.05±0.321.02±0.18
    单纯Vγ4T细胞清除组31.72±0.100.83±0.201.76±0.4219.83±0.53
    对照+1次UVR组32.56±0.331.24±0.172.94±0.2657.92±8.78
    Vγ4T细胞清除+1次UVR组31.00±0.190.22±0.060.33±0.117.71±0.33
    t13.670.841.9247.40
    P10.0220.4470.127<0.001
    t25.300.876.509.16
    P20.0060.4350.003<0.001
    t34.574.134.6727.36
    P30.0100.0150.010<0.001
    t45.778.1812.908.08
    P40.0050.001<0.0010.001
    注:NKG2D为自然杀伤细胞2族成员D,UVR为紫外线辐射,H60为组织相容性抗原60,Mult1为小鼠UL16结合蛋白样转录子1,Rae1为维甲酸早期诱导蛋白1;对单纯Vγ4T细胞清除组、单纯对照组小鼠分别予以腹腔内注射亚美尼亚仓鼠抗小鼠Vγ4TCR单克隆抗体、同型对照IgG抗体,并于注射后1周进行检测,对对照+1次UVR组和Vγ4T细胞清除+1次UVR组小鼠分别同单纯对照组、单纯Vγ4T细胞清除组注射后1周均另行1次UVR,然后进行检测;NKG2D、H60、Mult1、Rae1的mRNA表达的Vγ4T细胞清除主效应,F值分别为7.75、2.49、0.95、50.65,P值分别为0.050、0.190、0.386、0.002;UVR主效应,F值分别为5.77、17.17、28.82、25.95,P值分别为0.074、0.014、0.006、0.007;两者交互作用,F值分别为58.08、10.04、47.51、120.30,P值分别为0.002、0.034、0.002、<0.001;t1值、P1值为单纯对照组与单纯Vγ4T细胞清除组比较所得;t2值、P2值为单纯对照组与对照+1次UVR组比较所得;t3值、P3值为单纯Vγ4T细胞清除组与Vγ4T细胞清除+1次UVR组比较所得,t4值、P4值为对照+1次UVR组与Vγ4T细胞清除+1次UVR组比较所得
    下载: 导出CSV
  • 加载中
图(4) / 表(2)
计量
  • 文章访问数:  86
  • HTML全文浏览量:  34
  • PDF下载量:  8
  • 被引次数: 0
出版历程
  • 收稿日期:  2024-01-21

目录

    /

    返回文章
    返回