T细胞亚群与CD4~+ CD25~+调节性T细胞及Foxp3 mRNA在肾移植大鼠急性排斥反应中的作用
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摘要
目的探讨大鼠肾移植术后急性排斥反应中外周血T淋巴细胞亚群、CD4~+CD25~+调节性T细胞比率和叉头状转录因子p3(forkhead transcription factor p3,Foxp3)mRNA表达水平及意义。方法 SD大鼠15只为对照组;SD大鼠15只为受体、SD大鼠15只为供体建立同系基因移植非急性排斥组;SD大鼠15只为受体、Wistar大鼠15只为供体建立非同系基因移植急性排斥组。于肾移植术后第7天检测3组受体大鼠外周血T淋巴细胞亚群、CD4~+CD25~+调节性T细胞比率、Foxp3 mRNA水平及血清肌酐、尿素氮水平,对移植肾行组织病理学检査并进行肾移植急性排斥反应分级,并进行比较。结果肾脏组织病理显示,对照组大鼠无明显病理变化;非急性排斥组4只受体大鼠呈现交界性改变,小灶状间质炎性细胞浸润,3只出现急性排斥反应Ⅰ级病变,其余8只受体大鼠未见明显病理变化;急性排斥组大鼠肾脏呈急性排斥反应典型病理改变,可见交界性改变、灶状间质炎性细胞浸润、多灶状间质炎性细胞浸润、弥漫状间质炎性细胞浸润病理变化,其中急性排斥反应Ⅰ级2例、Ⅱ级2例、Ⅲ级5例、Ⅲb级6例。术后第7天急性排斥组血清肌酐[(123.50±8.06)μmol/L]、尿素氮[(7.40±0.75)mmol/L]水平高于非急性排斥组[(107.99±5.73)μmol/L、(6.04±0.84)mmol/L]和对照组[(49.28±6.85)μmol/L、(3.02±0.67)mmol/L](P<0.05);急性排斥组CD3~+T细胞比率[(50.68±7.59)%]、CD4~+T细胞比率[(35.81±5.49)%]、CD4~+/CD8~+(2.80±0.22)高于非急性排斥组[(41.97±9.01)%、(24.89±6.04)%、1.61±0.14]和对照组[(39.07±10.91)%、(21.74±6.02)%、1.43±0.14](P<0.05);急性排斥组CD8~+T细胞比率[(11.01±3.30)%]低于非急性排斥组[(15.39±4.07)%]和对照组[(15.61±4.81)%],CD4~+CD25~+调节性T细胞比率[(1.30±0.20)%]与Foxp3 mRNA[(0.33±0.08)×10~(-3)]水平低于非急性排斥组[(2.90±0.40)%、(1.02±0.21)×10~(-3)]和对照组[(4.97±0.50)%、(2.13±0.26)×10~(-3)](P<0.05)。结论外周血T细胞亚群参与调控肾移植术后急性排斥反应,急性排斥反应的发生可能与Foxp3 mRNA及CD4~+CD25~+调节性T细胞比率降低有关,Foxp3 mRNA及CD4~+CD25~+调节性T细胞可作为评估受体免疫状态、判断免疫耐受的参考指标。
Objective To investigate the expressions and significances of T lymphocyte subsets,CD25~+ CD4~+ regulatory T cells and forkhead transcription factor p3(Foxp3) mRNA in the peripheral blood of the rats with acute rejection after kidney transplantation.Methods T lymphocyte subsets,CD4~+ CD25~+ regulatory T cells rate,Foxp3 mRNA level,creatinine and blood urea nitrogen were detected on postoperative day 7 in acute rejection group(15 SD rats as recipients and 15 Wistar rats as donors),non-acute rejection group(15 SD rats as recipients and 15 SD rats as donors) and control group(15 SD rats),and the acute rejection of renal tissues was detected and graded by histopathological examination,and was compared.Results There was no obvious pathological change in control group.Four recipients showed borderline change,focal necrosis and interstitial inflammatory cell infiltration,and 3 recipients had grade I acute rejection,other 8recipients had no obvious pathological change in non-acute rejection group.The kidney in acute rejection group showed typical pathological changes of acute rejection,as borderline change,focal interstitial infiltration of inflammatory cells,multifocal interstitial infiltration of inflammatory cells,diffuse interstitial infiltration of inflammatory cells,including grade I acute rejection in 2 cases,grade Ⅱ in 2,grade Ⅲ a in 5 and grade Ⅲ b in 6.The levels of serum creatinine((123.50±8.06) μmol/L) and urea nitrogen((7.40±0.75) mmol/L) on postoperative day 7 in acute rejection group were significantly higher than those in non-acute rejection group((107.99±5.73) μmol/L,(6.04±0.84) mmol/L) and control group((49.28±6.85) μmol/L,(3.02±0.67) mmol/L)(P<0.05).The rates of CD3~+ T cell((50.68±7.59)%),CD4~+ T cell((35.81±5.49)%) and CD4~+/CD8~+(2.80±0.22) in acute rejection group were significantly higher than those in non-acute rejection group((41.97±9.01)%,(24.89±6.04)%,1.61±0.14) and control group((39.07±10.91)%,(21.74±6.02)%,1.43±0.14)(P<0.05),and the rate of CD8~+ T cell((11.01±3.30)%) in acute rejection group was significantly lower than that in non-acute rejection group((15.39±4.07)%) and control group((15.61±4.81)%)(P<0.05).The rates of CD4~+CD25~+ regulatory T cell((1.30±0.20)%) and the level of Foxp3mRNA((0.33±0.08)×10~(-3)) in acute rejection group were significantly lower than those in non-acute rejection group((2.90±0.40)%,(1.02±0.21)×10~(-3)) and control group((4.97±0.50)%,(2.13±0.26)×10~(-3))(P<0.05).Conclusion The peripheral blood T cell subsets participate in the regulation of acute rejection after kidney transplantation.The occurrence of acute rejection may be correlated with the decrease of Foxp3 mRNA and CD4~+CD25~+regulatory T cells.The rate of CD4~+ CD25~+ regulatory T cells and the expression level of Foxp3 mRNA in peripheral blood after transplantation could be used as the reference indexes to evaluate the immune status of the recipient and to judge the immune tolerance.
Objective To investigate the expressions and significances of T lymphocyte subsets,CD25~+ CD4~+ regulatory T cells and forkhead transcription factor p3(Foxp3) mRNA in the peripheral blood of the rats with acute rejection after kidney transplantation.Methods T lymphocyte subsets,CD4~+ CD25~+ regulatory T cells rate,Foxp3 mRNA level,creatinine and blood urea nitrogen were detected on postoperative day 7 in acute rejection group(15 SD rats as recipients and 15 Wistar rats as donors),non-acute rejection group(15 SD rats as recipients and 15 SD rats as donors) and control group(15 SD rats),and the acute rejection of renal tissues was detected and graded by histopathological examination,and was compared.Results There was no obvious pathological change in control group.Four recipients showed borderline change,focal necrosis and interstitial inflammatory cell infiltration,and 3 recipients had grade I acute rejection,other 8recipients had no obvious pathological change in non-acute rejection group.The kidney in acute rejection group showed typical pathological changes of acute rejection,as borderline change,focal interstitial infiltration of inflammatory cells,multifocal interstitial infiltration of inflammatory cells,diffuse interstitial infiltration of inflammatory cells,including grade I acute rejection in 2 cases,grade Ⅱ in 2,grade Ⅲ a in 5 and grade Ⅲ b in 6.The levels of serum creatinine((123.50±8.06) μmol/L) and urea nitrogen((7.40±0.75) mmol/L) on postoperative day 7 in acute rejection group were significantly higher than those in non-acute rejection group((107.99±5.73) μmol/L,(6.04±0.84) mmol/L) and control group((49.28±6.85) μmol/L,(3.02±0.67) mmol/L)(P<0.05).The rates of CD3~+ T cell((50.68±7.59)%),CD4~+ T cell((35.81±5.49)%) and CD4~+/CD8~+(2.80±0.22) in acute rejection group were significantly higher than those in non-acute rejection group((41.97±9.01)%,(24.89±6.04)%,1.61±0.14) and control group((39.07±10.91)%,(21.74±6.02)%,1.43±0.14)(P<0.05),and the rate of CD8~+ T cell((11.01±3.30)%) in acute rejection group was significantly lower than that in non-acute rejection group((15.39±4.07)%) and control group((15.61±4.81)%)(P<0.05).The rates of CD4~+CD25~+ regulatory T cell((1.30±0.20)%) and the level of Foxp3mRNA((0.33±0.08)×10~(-3)) in acute rejection group were significantly lower than those in non-acute rejection group((2.90±0.40)%,(1.02±0.21)×10~(-3)) and control group((4.97±0.50)%,(2.13±0.26)×10~(-3))(P<0.05).Conclusion The peripheral blood T cell subsets participate in the regulation of acute rejection after kidney transplantation.The occurrence of acute rejection may be correlated with the decrease of Foxp3 mRNA and CD4~+CD25~+regulatory T cells.The rate of CD4~+ CD25~+ regulatory T cells and the expression level of Foxp3 mRNA in peripheral blood after transplantation could be used as the reference indexes to evaluate the immune status of the recipient and to judge the immune tolerance.
引文
[1]王毅,陈正,熊烈,等.青藤碱对肾移植大鼠急性排斥反应及T细胞增殖的影响[J].中华实验外科杂志,2004,21(5):573-574.
[2]Karczewski M,Karczewski J,Kostrzewa A,et al.The role of Foxp3+regulatory T cells in kidney transplantation[J].Transplant Proc,2009,41(5):1527-1529.
[3]韩永,蔡明,郭晖,等.移植肾术后急性抗体介导性排斥反应的诊断与治疗[J].中华实用诊断与治疗杂志,2015,29(1):34-36.
[4]张艾莎,刘新华,陈继红,等.大鼠肾移植后外周血T细胞亚群动态变化[J].中华实用诊断与治疗杂志,2015,29(11):1095-1097.
[5]陈继红,热衣汗·西里普,刘健,等.大鼠肾移植急性排斥模型建立研究[J].中华实用诊断与治疗杂志,2012,26(4):355-357.
[6]Solez K,Axelsen RA,Benediktsson H,et al.International standardization of criteria for histologic diagnosis of renal allograft rejection,the Banff working classification of kidney transplant pathology[J].Kidney Int,1993,44(2):411-422.
[7]田军,张金元,胡大勇,等.单剂量巴利昔单抗对肾移植受者外周血Foxp3mRNA、CD4~+CD25~+调节性T细胞、白细胞介素-2及其受体的影响[J].中国组织工程研究,2007,11(25):4861-4865.
[8]Newell KA,Larsen CP.Toward transplantation tolerance:a large step on a long road[J].Am J Transplant,2006,6(9):1989-1990.
[9]Ducloux D,Courivaud C,Bamoulid J,et al.Prolonged CD4+T cell lymphopenia increases morbidity and mortality after renal transplantation[J].J Am Soc Nephrol,2010,21(5):868-875.
[10]Issa F,Schiopu A,Wood KJ.Role of T cells in graft rejection and transplantation tolerance[J].Expert Rev Clin Immunol,2010,6(1):155-169.
[11]张翠芳.T淋巴细胞在临床疾病发病与诊治中的作用探讨[J].中国中医药咨讯,2011,3(19):219.
[12]熊海云,张雷,王立明,等.肾移植后外周血CD4~+及CD8~+T细胞亚群计数的临床意义[J].中国组织工程研究,2011,15(53):9957-9960.
[13]Wever PC,Spaeny LH,van der Vliet HJ,et al.Expression of granzyme B during primary cytomegalovirus infection after renal transplantation[J].J Infect Dis,1999,179(3):693-696.
[14]Sakaguchi S,Setoguchi R,Yagi H,et al.Naturally arising Foxp3-expressing CD25~+CD4~+regulatory T cells in selftolerance and autoimmune disease[J].Curr Top Microbiol Immunol,2006,305:51-66.
[15]Yamazaki S,Patel M,Harper A,et al.Effective expansion of alloantigerrspecific Foxp3~+CD25~+CD4~+regulatory T cells by dendritic cells during the mixed leukocyte reaction[J].Proc Natl Acad Sci USA,2006,103(8):2758-2763.
[16]Braudeau C,Racape M,Giral M,et al.Variation in numbers of CD4~+CD25 high Foxp3~+T cells with normal immuncr regulatory properties in long-term graft outcome[J].Trans pi Int,2007,20(10):845-855.
[17]罗文峰,时军,黄跃胜,等.调节性T细胞与肾移植急性排斥反应的关系[J].江西医药,2014,49(1):9-11.
[18]李川江,于立新,付绍杰,等.移植肾急性排斥患者外周血调节性T细胞Foxp3的动态监测[J].实用医学杂志,2011,27(22):4065-4067.
[19]王旭珍,薛武军,田晓辉,等.肾移植后淋巴细胞亚群的变化及其对诊断急性排斥反应和CMV感染的意义[J].中华器官移植杂志,2013,34(11):651-654.
[2]Karczewski M,Karczewski J,Kostrzewa A,et al.The role of Foxp3+regulatory T cells in kidney transplantation[J].Transplant Proc,2009,41(5):1527-1529.
[3]韩永,蔡明,郭晖,等.移植肾术后急性抗体介导性排斥反应的诊断与治疗[J].中华实用诊断与治疗杂志,2015,29(1):34-36.
[4]张艾莎,刘新华,陈继红,等.大鼠肾移植后外周血T细胞亚群动态变化[J].中华实用诊断与治疗杂志,2015,29(11):1095-1097.
[5]陈继红,热衣汗·西里普,刘健,等.大鼠肾移植急性排斥模型建立研究[J].中华实用诊断与治疗杂志,2012,26(4):355-357.
[6]Solez K,Axelsen RA,Benediktsson H,et al.International standardization of criteria for histologic diagnosis of renal allograft rejection,the Banff working classification of kidney transplant pathology[J].Kidney Int,1993,44(2):411-422.
[7]田军,张金元,胡大勇,等.单剂量巴利昔单抗对肾移植受者外周血Foxp3mRNA、CD4~+CD25~+调节性T细胞、白细胞介素-2及其受体的影响[J].中国组织工程研究,2007,11(25):4861-4865.
[8]Newell KA,Larsen CP.Toward transplantation tolerance:a large step on a long road[J].Am J Transplant,2006,6(9):1989-1990.
[9]Ducloux D,Courivaud C,Bamoulid J,et al.Prolonged CD4+T cell lymphopenia increases morbidity and mortality after renal transplantation[J].J Am Soc Nephrol,2010,21(5):868-875.
[10]Issa F,Schiopu A,Wood KJ.Role of T cells in graft rejection and transplantation tolerance[J].Expert Rev Clin Immunol,2010,6(1):155-169.
[11]张翠芳.T淋巴细胞在临床疾病发病与诊治中的作用探讨[J].中国中医药咨讯,2011,3(19):219.
[12]熊海云,张雷,王立明,等.肾移植后外周血CD4~+及CD8~+T细胞亚群计数的临床意义[J].中国组织工程研究,2011,15(53):9957-9960.
[13]Wever PC,Spaeny LH,van der Vliet HJ,et al.Expression of granzyme B during primary cytomegalovirus infection after renal transplantation[J].J Infect Dis,1999,179(3):693-696.
[14]Sakaguchi S,Setoguchi R,Yagi H,et al.Naturally arising Foxp3-expressing CD25~+CD4~+regulatory T cells in selftolerance and autoimmune disease[J].Curr Top Microbiol Immunol,2006,305:51-66.
[15]Yamazaki S,Patel M,Harper A,et al.Effective expansion of alloantigerrspecific Foxp3~+CD25~+CD4~+regulatory T cells by dendritic cells during the mixed leukocyte reaction[J].Proc Natl Acad Sci USA,2006,103(8):2758-2763.
[16]Braudeau C,Racape M,Giral M,et al.Variation in numbers of CD4~+CD25 high Foxp3~+T cells with normal immuncr regulatory properties in long-term graft outcome[J].Trans pi Int,2007,20(10):845-855.
[17]罗文峰,时军,黄跃胜,等.调节性T细胞与肾移植急性排斥反应的关系[J].江西医药,2014,49(1):9-11.
[18]李川江,于立新,付绍杰,等.移植肾急性排斥患者外周血调节性T细胞Foxp3的动态监测[J].实用医学杂志,2011,27(22):4065-4067.
[19]王旭珍,薛武军,田晓辉,等.肾移植后淋巴细胞亚群的变化及其对诊断急性排斥反应和CMV感染的意义[J].中华器官移植杂志,2013,34(11):651-654.