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Volume 38 Issue 5
May  2022
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2022  >  38(5): 462-470
Liu ZY,Cheng X,Zhang JX,et al.Role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice[J].Chin J Burns Wounds,2022,38(5):462-470.DOI: 10.3760/cma.j.cn501120-20201209-00523.
Citation: Liu ZY,Cheng X,Zhang JX,et al.Role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice[J].Chin J Burns Wounds,2022,38(5):462-470.DOI: 10.3760/cma.j.cn501120-20201209-00523.
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Role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice

doi: 10.3760/cma.j.cn501120-20201209-00523

  • Medical Cosmetic Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China

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  • Corresponding author: Shi Ke, Email: 822870715@qq.com
  • Received Date: 2020-12-09
    Abstract
  •     Objective   To investigate the role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice.    Methods   The experimental research methods were applied. Eighty-six C57BL/6J male mice (hereinafter briefly referred to as wild-type mice) aged 8-12 weeks were selected for the following experiments. Vγ4 T cells were isolated from axillary lymph nodes of five wild-type mice for the following experiments. Intraperitoneal injection of rapamycin for 42 mice was performed to establish rapamycin-treated mice model for the following experiments. Eighteen wild-type mice were divided into normal control group without any treatment, trauma only group, and trauma+CC chemokine ligand 20 (CCL20) inhibitor group according to the random number table (the same grouping method below), with 6 mice in each group. The full-thickness skin defect wound was made on the back of mice in the latter two groups (the same wound model below), and mice in trauma+CCL20 inhibitor group were continuously injected subcutaneously with CCL20 inhibitor at the wound edge for 3 days after injury. Another 6 rapamycin-treated mice were used to establish wound model as rapamycin+trauma group. On post injury day (PID) 3, the epidermal cells of the skin tissue around the wound of each trauma mice were extracted by enzyme digestion, and the percentage of Vγ4 T cells in the epidermal cells was detected by flow cytometry. In normal control group, the epidermal cells of the normal skin tissue in the back of mice were taken at the appropriate time point for detection as above. Five wild-type mice were used to establish wound models. On PID 3, the epidermal cells were extracted from the skin tissue around the wound. The cell populations were divided into Vγ4 T cells, Vγ3 T cells, and γδ negative cells by fluorescence-activated cell sorter, which were set as Vγ4 T cell group, Vγ3 T cell group, and γδ negative cell group (with cells in each group being mixed with B16 mouse melanoma cells), respectively. B16 mouse melanoma cells were used as melanoma cell control group. The expression of interleukin-22 (IL-22) mRNA in cells of each group was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with the number of samples being 6. Thirty rapamycin-treated mice were used to establish wound models, which were divided into Vγ4 T cell only group and Vγ4 T cell+IL-22 inhibitor group performed with corresponding injections and rapamycin control group injected with phosphate buffer solution (PBS) immediately after injury, with 10 mice in each group. Another 10 wild-type mice were taken to establish wound models and injected with PBS as wild-type control group. Mice in each group were injected continuously for 6 days. The percentage of wound area of mice in the four groups was calculated on PID 1, 2, 3, 4, 5, and 6 after injection on the same day. Six wild-type mice and 6 rapamycin-treated mice were taken respectively to establish wound models as wild-type group and rapamycin group. On PID 3, the mRNA and protein expressions of IL-22 and CCL20 in the peri-wound epidermis tissue of mice in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The Vγ4 T cells were divided into normal control group without any treatment and rapamycin-treated rapamycin group. After being cultured for 24 hours, the mRNA and protein expressions of IL-22 of cells in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively, with the number of samples being 6. Data were statistically analyzed with independent sample t test, analysis of variance for repeated measurement, one-way analysis of variance, Bonferroni method, Kruskal-Wallis H test, and Wilcoxon rank sum test.    Results   The percentage of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in trauma only group on PID 3 was 0.66% (0.52%, 0.81%), which was significantly higher than 0.09% (0.04%, 0.14%) in the epidermal cells of the normal skin tissue of mice in normal control group (Z=4.31, P<0.01). The percentages of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in rapamycin+trauma group and trauma+CCL20 inhibitor group on PID 3 were 0.25% (0.16%, 0.37%) and 0.24% (0.17%, 0.35%), respectively, which were significantly lower than that in trauma only group (with Z values of 2.27 and 2.25, respectively, P<0.05). The mRNA expression level of IL-22 of cells in Vγ4 T cell group was significantly higher than that in Vγ3 T cell group, γδ negative cell group, and melanoma cell control group (with Z values of 2.96, 2.45, and 3.41, respectively, P<0.05 or P<0.01). Compared with that in wild-type control group, the percentage of wound area of mice in rapamycin control group increased significantly on PID 1-6 (P<0.01), the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 1 and PID 3-6 (P<0.05 or P<0.01). Compared with that in rapamycin control group, the percentage of wound area of mice in Vγ4 T cell only group decreased significantly on PID 1-6 (P<0.05 or P<0.01). Compared with that in Vγ4 T cell only group, the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 3-6 (P<0.05 or P<0.01). On PID 3, compared with those in wild-type group, the expression levels of IL-22 protein and mRNA (with t values of -7.82 and -5.04, respectively, P<0.01) and CCL20 protein and mRNA (with t values of -7.12 and -5.73, respectively, P<0.01) were decreased significantly in the peri-wound epidermis tissue of mice in rapamycin group. After being cultured for 24 hours, the expression levels of IL-22 protein and mRNA in Vγ4 T cells in rapamycin group were significantly lower than those in normal control group (with t values of -7.75 and -6.04, respectively, P<0.01).    Conclusions   In mice with full-thickness skin defects, rapamycin may impair the CCL20 chemotactic system by inhibiting the expression of CCL20, leading to a decrease in the recruitment of Vγ4 T cells to the epidermis, and at the same time inhibit the secretion of IL-22 by Vγ4 T cells, thereby slowing the wound healing rate.

     

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    • References(35)
  • [1]
    SehgalSN.Sirolimus: its discovery, biological properties, and mechanism of action[J].Transplant Proc,2003,35(3 Suppl):S7-14.DOI: 10.1016/s0041-1345(03)00211-2.
    [2]
    HymesLC,WarshawBL.Sirolimus in pediatric patients: results in the first 6 months post-renal transplant[J].Pediatr Transplant,2005,9(4):520-522.DOI: 10.1111/j.1399-3046.2005.00324.x.
    [3]
    SchäfferM,SchierR,NapireiM,et al.Sirolimus impairs wound healing[J].Langenbecks Arch Surg,2007,392(3):297-303.DOI: 10.1007/s00423-007-0174-5.
    [4]
    MacDonaldAS.Rapamycin in combination with cyclosporine or tacrolimus in liver, pancreas, and kidney transplantation[J].Transplant Proc,2003,35(3 Suppl):S201-208.DOI: 10.1016/s0041-1345(03)00231-8.
    [5]
    KuppahallyS,Al-KhaldiA,WeisshaarD,et al.Wound healing complications with de novo sirolimus versus mycophenolate mofetil-based regimen in cardiac transplant recipients[J].Am J Transplant,2006,6(5 Pt 1):986-992.DOI: 10.1111/j.1600-6143.2006.01282.x.
    [6]
    DeanPG,LundWJ,LarsonTS,et al.Wound-healing complications after kidney transplantation: a prospective, randomized comparison of sirolimus and tacrolimus[J].Transplantation,2004,77(10):1555-1561.DOI: 10.1097/01.tp.0000123082.31092.53.
    [7]
    ValenteJF,HricikD,WeigelK,et al.Comparison of sirolimus vs. mycophenolate mofetil on surgical complications and wound healing in adult kidney transplantation[J].Am J Transplant,2003,3(9):1128-1134.DOI: 10.1034/j.1600-6143.2003.00185.x.
    [8]
    MillsRE,TaylorKR,PodshivalovaK,et al.Defects in skin gamma delta T cell function contribute to delayed wound repair in rapamycin-treated mice[J].J Immunol,2008,181(6):3974-3983.DOI: 10.4049/jimmunol.181.6.3974.
    [9]
    HaidingerM,PoglitschM,GeyereggerR,et al.A versatile role of mammalian target of rapamycin in human dendritic cell function and differentiation[J].J Immunol,2010,185(7):3919-3931.DOI: 10.4049/jimmunol.1000296.
    [10]
    YanezDA,LacherRK,VidyarthiA,et al.The role of macrophages in skin homeostasis[J].Pflugers Arch,2017,469(3/4):455-463.DOI: 10.1007/s00424-017-1953-7.
    [11]
    DiegelmannRF,EvansMC.Wound healing: an overview of acute, fibrotic and delayed healing[J].Front Biosci,2004,9:283-289.DOI: 10.2741/1184.
    [12]
    SutohY,MohamedRH,KasaharaM.Origin and evolution of dendritic epidermal T cells[J].Front Immunol,2018,9:1059.DOI: 10.3389/fimmu.2018.01059.
    [13]
    ChenC,MengZ,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[2022-03-22]. https://pubmed.ncbi.nlm.nih.gov/34212060/.DOI: 10.1093/burnst/tkab009.
    [14]
    MarshallAS,SilvaJR,BannermanCA,et al.Skin-resident γδ T cells exhibit site-specific morphology and activation states[J].J Immunol Res,2019,2019:9020234.DOI: 10.1155/2019/9020234.
    [15]
    LiuZ,XuY,ZhangX,et al.Defects in dermal Vγ4 γ δ T cells result in delayed wound healing in diabetic mice[J].Am J Transl Res,2016,8(6):2667-2680.
    [16]
    MabuchiT,TakekoshiT,HwangST.Epidermal CCR6+ γδ T cells are major producers of IL-22 and IL-17 in a murine model of psoriasiform dermatitis[J].J Immunol,2011,187(10):5026-5031.DOI: 10.4049/jimmunol.1101817.
    [17]
    McGeeHM,SchmidtBA,BoothCJ,et al.IL-22 promotes fibroblast-mediated wound repair in the skin[J].J Invest Dermatol,2013,133(5):1321-1329.DOI: 10.1038/jid.2012.463.
    [18]
    AugustineJJ,BodziakKA,HricikDE.Use of sirolimus in solid organ transplantation[J].Drugs,2007,67(3):369-391.DOI: 10.2165/00003495-200767030-00004.
    [19]
    RuchinPE,MullerDW,FaddySC,et al.Long-term clinical follow-up of sirolimus-eluting (CYPHER) coronary stents in the treatment of instent restenosis in an unselected population[J].Heart Lung Circ,2007,16(6):440-446.DOI: 10.1016/j.hlc.2007.02.090.
    [20]
    WitzigTE,KaufmannSH.Inhibition of the phosphatidylinositol 3-kinase/mammalian target of rapamycin pathway in hematologic malignancies[J].Curr Treat Options Oncol,2006,7(4):285-294.DOI: 10.1007/s11864-006-0038-1.
    [21]
    PhungTL,ZivK,DabydeenD,et al.Pathological angiogenesis is induced by sustained Akt signaling and inhibited by rapamycin[J].Cancer Cell,2006,10(2):159-170.DOI: 10.1016/j.ccr.2006.07.003.
    [22]
    贺伟峰.皮肤γδT细胞各亚群在创面再上皮化过程中的调控作用及其相关机制[J].中华烧伤与创面修复杂志,2022,38(2):114-118.DOI: 10.3760/cma.j.cn501120-20211210-00411.
    [23]
    刘勉,朱海杰,杨加彩,等.树突状表皮T淋巴细胞对小鼠创缘表皮细胞增殖和凋亡的影响[J].中华烧伤杂志,2020,36(2):122-130.DOI: 10.3760/cma.j.issn.1009-2587.2020.02.008.
    [24]
    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.
    [25]
    ZhengY,CollinsSL,LutzMA,et al.A role for mammalian target of rapamycin in regulating T cell activation versus anergy[J].J Immunol,2007,178(4):2163-2170.DOI: 10.4049/jimmunol.178.4.2163.
    [26]
    SchutyserE,StruyfS,Van DammeJ.The CC chemokine CCL20 and its receptor CCR6[J].Cytokine Growth Factor Rev,2003,14(5):409-426.DOI: 10.1016/s1359-6101(03)00049-2.
    [27]
    HomeyB,Dieu-NosjeanMC,WiesenbornA,et al.Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis[J].J Immunol,2000,164(12):6621-6632.DOI: 10.4049/jimmunol.164.12.6621.
    [28]
    MabuchiT,SinghTP,TakekoshiT,et al.CCR6 is required for epidermal trafficking of γδ-T cells in an IL-23-induced model of psoriasiform dermatitis[J].J Invest Dermatol,2013,133(1):164-171.DOI: 10.1038/jid.2012.260.
    [29]
    FurueK,ItoT,TsujiG,et al.The CCL20 and CCR6 axis in psoriasis[J].Scand J Immunol,2020,91(3):e12846.DOI: 10.1111/sji.12846.
    [30]
    FujitaH.The role of IL-22 and Th22 cells in human skin diseases[J].J Dermatol Sci,2013,72(1):3-8.DOI: 10.1016/j.jdermsci.2013.04.028.
    [31]
    MiyagakiT,SugayaM,SugaH,et al.IL-22, but not IL-17, dominant environment in cutaneous T-cell lymphoma[J].Clin Cancer Res,2011,17(24):7529-7538.DOI: 10.1158/1078-0432.CCR-11-1192.
    [32]
    AvitabileS,OdorisioT,MadonnaS,et al.Interleukin-22 promotes wound repair in diabetes by improving keratinocyte pro-healing functions[J].J Invest Dermatol,2015,135(11):2862-2870.DOI: 10.1038/jid.2015.278.
    [33]
    KolumamG,WuX,LeeWP,et al.IL-22R ligands IL-20, IL-22, and IL-24 promote wound healing in diabetic db/db mice[J].PLoS One,2017,12(1):e0170639.DOI: 10.1371/journal.pone.0170639.
    [34]
    TillackC,EhmannLM,FriedrichM,et al.Anti-TNF antibody- induced psoriasiform skin lesions in patients with inflammatory bowel disease are characterised by interferon-γ-expressing Th1 cells and IL-17A/IL-22-expressing Th17 cells and respond to anti-IL-12/IL-23 antibody treatment[J].Gut,2014,63(4):567-577.DOI: 10.1136/gutjnl-2012-302853.
    [35]
    Van BelleAB,de HeuschM,LemaireMM,et al.IL-22 is required for imiquimod-induced psoriasiform skin inflammation in mice[J].J Immunol,2012,188(1):462-469.DOI: 10.4049/jimmunol.1102224.
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