中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2013年
11期
680-684
,共5页
李庆山%陈钊%杜庆华%邓家德%凌艳英%江雪杰%孟凡义
李慶山%陳釗%杜慶華%鄧傢德%凌豔英%江雪傑%孟凡義
리경산%진쇠%두경화%산가덕%릉염영%강설걸%맹범의
粒细胞集落刺激因子,重组%T淋巴细胞,调节%白细胞介素17%造血干细胞移植
粒細胞集落刺激因子,重組%T淋巴細胞,調節%白細胞介素17%造血榦細胞移植
립세포집락자격인자,중조%T림파세포,조절%백세포개소17%조혈간세포이식
Granulocyte colony stimulating factor,recombinant%Interleukin-17%T-Lymphocytes,regulatory%Hematopoietic stem cell transplantation
目的 探讨重组人粒细胞集落刺激因子(rhG-CSF)对造血干细胞移植供者外周血中TH17细胞和Treg细胞的影响及其对CD4+T淋巴细胞中细胞因子转导抑制因子3(SOCS3)基因表达的影响.方法 16名供者连续5d皮下注射rhG-CSF,5μg·kg-1·d-1,进行外周血造血干细胞动员,采集动员第0、3、5天外周血及造血干细胞移植物中单个核细胞和血清,采用流式细胞术检测单个核细胞中TH17细胞和Treg细胞比例,酶联免疫吸附试验法检测血清中白细胞介素17A(IL-17A)、白细胞介素(IL-21)、白细胞介素23(IL-23)和转化生长因子β1(TGF-β1)浓度,免疫磁珠分选出单个核中CD4+T淋巴细胞,采用实时定量PCR技术检测CD4+T淋巴细胞中细胞因子信号转导抑制因子(SOCS3)基因的表达.结果 动员第0、3、5天外周血及干细胞采集物中,TH 17细胞比例分别为(2.69±0.81)％、(0.91±0.33)％、(0.35±0.12)％、(0.21±0.05)％,逐渐下降(P＜0.05);Treg细胞比例分别为(0.56±0.24)％、(0.72±0.22)％、(1.59±0.54)％、(3.38±0.52)％,逐渐上升(P＜0.05).动员第0、3、5天,IL-17A浓度分别为(7.33±0.89)μg/L、(5.78±1.03) μg/L、(3.32±0.84) μg/L;IL-21浓度分别为(124.56±15.18) μg/L、(117.12±14.45)μ.g/L、(64.94±11.25) μg/L; TGF-β1浓度分别为(183.52±59.35)μg/L、(280.49±69.75) μg/L、(393.62±57.25)μg/L;IL-23浓度分别为(45.89±6.95)μg/L、(46.25±7.44)μg/L、(47.45±10.75) μg/L.IL-17A、IL-21浓度逐渐下降(P＜0.01),而TGF-β1浓度逐渐上升(P＜0.01),IL-23浓度无明显变化(P＞0.05).rhG-CSF动员后及干细胞采集物中CD4+T淋巴细胞内SOCS3基因的表达呈升高趋势.结论 rhG-CSF抑制外周血中TH17细胞产生,促进Treg细胞产生;降低血清中IL-17A、IL-21浓度,提高TGF-β1浓度,有利于CD4+T淋巴细胞向Treg细胞分化,与rhG-CSF促进CD4+T淋巴细胞中SOCS3基因的表达有关.
目的 探討重組人粒細胞集落刺激因子(rhG-CSF)對造血榦細胞移植供者外週血中TH17細胞和Treg細胞的影響及其對CD4+T淋巴細胞中細胞因子轉導抑製因子3(SOCS3)基因錶達的影響.方法 16名供者連續5d皮下註射rhG-CSF,5μg·kg-1·d-1,進行外週血造血榦細胞動員,採集動員第0、3、5天外週血及造血榦細胞移植物中單箇覈細胞和血清,採用流式細胞術檢測單箇覈細胞中TH17細胞和Treg細胞比例,酶聯免疫吸附試驗法檢測血清中白細胞介素17A(IL-17A)、白細胞介素(IL-21)、白細胞介素23(IL-23)和轉化生長因子β1(TGF-β1)濃度,免疫磁珠分選齣單箇覈中CD4+T淋巴細胞,採用實時定量PCR技術檢測CD4+T淋巴細胞中細胞因子信號轉導抑製因子(SOCS3)基因的錶達.結果 動員第0、3、5天外週血及榦細胞採集物中,TH 17細胞比例分彆為(2.69±0.81)％、(0.91±0.33)％、(0.35±0.12)％、(0.21±0.05)％,逐漸下降(P＜0.05);Treg細胞比例分彆為(0.56±0.24)％、(0.72±0.22)％、(1.59±0.54)％、(3.38±0.52)％,逐漸上升(P＜0.05).動員第0、3、5天,IL-17A濃度分彆為(7.33±0.89)μg/L、(5.78±1.03) μg/L、(3.32±0.84) μg/L;IL-21濃度分彆為(124.56±15.18) μg/L、(117.12±14.45)μ.g/L、(64.94±11.25) μg/L; TGF-β1濃度分彆為(183.52±59.35)μg/L、(280.49±69.75) μg/L、(393.62±57.25)μg/L;IL-23濃度分彆為(45.89±6.95)μg/L、(46.25±7.44)μg/L、(47.45±10.75) μg/L.IL-17A、IL-21濃度逐漸下降(P＜0.01),而TGF-β1濃度逐漸上升(P＜0.01),IL-23濃度無明顯變化(P＞0.05).rhG-CSF動員後及榦細胞採集物中CD4+T淋巴細胞內SOCS3基因的錶達呈升高趨勢.結論 rhG-CSF抑製外週血中TH17細胞產生,促進Treg細胞產生;降低血清中IL-17A、IL-21濃度,提高TGF-β1濃度,有利于CD4+T淋巴細胞嚮Treg細胞分化,與rhG-CSF促進CD4+T淋巴細胞中SOCS3基因的錶達有關.
목적 탐토중조인립세포집락자격인자(rhG-CSF)대조혈간세포이식공자외주혈중TH17세포화Treg세포적영향급기대CD4+T림파세포중세포인자전도억제인자3(SOCS3)기인표체적영향.방법 16명공자련속5d피하주사rhG-CSF,5μg·kg-1·d-1,진행외주혈조혈간세포동원,채집동원제0、3、5천외주혈급조혈간세포이식물중단개핵세포화혈청,채용류식세포술검측단개핵세포중TH17세포화Treg세포비례,매련면역흡부시험법검측혈청중백세포개소17A(IL-17A)、백세포개소(IL-21)、백세포개소23(IL-23)화전화생장인자β1(TGF-β1)농도,면역자주분선출단개핵중CD4+T림파세포,채용실시정량PCR기술검측CD4+T림파세포중세포인자신호전도억제인자(SOCS3)기인적표체.결과 동원제0、3、5천외주혈급간세포채집물중,TH 17세포비례분별위(2.69±0.81)％、(0.91±0.33)％、(0.35±0.12)％、(0.21±0.05)％,축점하강(P＜0.05);Treg세포비례분별위(0.56±0.24)％、(0.72±0.22)％、(1.59±0.54)％、(3.38±0.52)％,축점상승(P＜0.05).동원제0、3、5천,IL-17A농도분별위(7.33±0.89)μg/L、(5.78±1.03) μg/L、(3.32±0.84) μg/L;IL-21농도분별위(124.56±15.18) μg/L、(117.12±14.45)μ.g/L、(64.94±11.25) μg/L; TGF-β1농도분별위(183.52±59.35)μg/L、(280.49±69.75) μg/L、(393.62±57.25)μg/L;IL-23농도분별위(45.89±6.95)μg/L、(46.25±7.44)μg/L、(47.45±10.75) μg/L.IL-17A、IL-21농도축점하강(P＜0.01),이TGF-β1농도축점상승(P＜0.01),IL-23농도무명현변화(P＞0.05).rhG-CSF동원후급간세포채집물중CD4+T림파세포내SOCS3기인적표체정승고추세.결론 rhG-CSF억제외주혈중TH17세포산생,촉진Treg세포산생;강저혈청중IL-17A、IL-21농도,제고TGF-β1농도,유리우CD4+T림파세포향Treg세포분화,여rhG-CSF촉진CD4+T림파세포중SOCS3기인적표체유관.
Objective To explore the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) mobilization on TH17/Treg cells and its impact on suppressor of cytokine signaling-3 (SOCS3) gene expression in CD4+ T cells in donors' peripheral blood.Method Sixteen donors were injected subcutaneously with rhG-CSF 5 μg/kg every day for 5 consecutive days for peripheral blood stem cells mobilization.At the first 0,3,5 day,the mononuclear cclls (MNCs) in peripheral blood or graft and serum specimens were taken.The CD4 + T cells in MNCs were sorted using immuno-magnetic beads.The ratio of TH 17 and Treg cells in MNCs,cytokines concentrations of IL-17A,IL-21,ID23 and TGFβ1 in serum,and SODC3 gene expression in CD4+ T cells were detected by using flow cytometry,ELISA,and reverse transcription real-time quantitative PCR (RT-qPCR),respectively.Results (1)The ratio of Th17 cells (CD3+ CD8 CD17+) and Treg cells (CD4+ CD25+ Foxp3+) in MNCs in peripheral blood and graft at the first 0,3 and 5 days after mobilization was (2.69 ± 0.81) ％,(0.91 ± 0.33) ％,(0.35 ± 0.12) ％,(0.21 ± 0.05) ％,and (0.56 ± 0.24) ％,(0.72 ± 0.22％),(1.59 ± 0.54) ％,(3.38 ± 0.52) ％,respectively,showing a significant declining and increasing trend respectively (P＜0.05); (2)The cytokine concentrations in serum at the first 0,3 and 5 days after mobilization were 7.33 ± 0.89,5.78 ± 1.03 and 3.32 ± 0.84 μg/L for IL-17A; 124.56 ± 15.18,117.12 ± 14.45 and 64.94 ± 11.25 μg/L for IL-21 ; 183.52 ± 59.35,280.49 ± 69.75 and 393.62 ± 57.25μg/L for TGF-β1 (P＜0.01) ; and 45.89 ± 6.95,46.25 ± 7.44 and 47.45 ± 10.75 μg/L for IL-23,respectively.The IL-17A and IL-21 concentrations showed significant declining trend,contrarily TGF-β1 with an increasing trend,while IL-23 concentration had no change.After rhG-CSF mobilization,the SOCS3 gene expression in CD4 + T cells of peripheral blood and graft at the first 0,3,5 days was gradually increased.Conclusion rhG-CSF suppresses Th17 cells and promotes regulatory T cells generation,meanwhile decreases IL-17A and IL-21 and elevates serum TGF-β1 concentrations,and contributes to CD4 + T cells differentiation to Tregs,probably by elevating SOSC3 gene expression in CD4+ T cells.
