注射供者的肝匀浆提取液延长大鼠异位心脏移植存活时间

目的探讨注射供者的肝匀浆提取液对大鼠淋巴细胞功能及大鼠异位移植心的影响。方法以Wistar大鼠为供者,SD大鼠为受者。制作Wistar大鼠的肝匀浆提取液；建立大鼠同种异体异位心脏移植模型。(1)经受者阴茎静脉注射肝匀浆提取液0．3 ml,14d后取供、受者的血液,用四甲基偶氮唑盐(MTT)法分别测定受者对同一供者和无关供者的单向混合淋巴细胞反应(MLR)。(2)心脏移植术前2h经受者阴茎静脉注射肝匀浆提取液0．3 ml。心脏移植术后分别观察受者注射同一供者和无关供者的肝匀浆提取液后移植心脏的存活时间；心脏停跳后取移植心做病理检查及免疫组织化学检测。结果受者对同一供者和无关供者的单向MLR比较,前者明显减轻,吸光度A值分别为：0．434±0．034和0．522±0．015,两组比较,差异有统计学意义(P＜0．01)。心脏移植术前,受者接受同一供者和无关供者的肝匀浆提取液后,前者移植心脏存活时间延长,分别为(38．05±17．07)d和(9．86±2．67)d,两组比较,差异有统计学意义(P＜0．01)；且前者心肌出血、坏死程度更轻,心肌组织内IgM和IgG沉积更少。结论注射同一供者的肝匀浆提取液能特异性抑制相应个体抗原引起的淋巴细胞增殖反应,减轻大鼠移植心脏的排斥反应,明显延长其存活时间。     

联合应用抗淋巴细胞血清和供体脾细胞延长大鼠移植心存活的作用

本实验主要观察术前短期用抗淋巴细胞血清（ＡＬＳ）；联合术前或术后输注供体脾细胞（ＤＳＣ），对移植心存活的作用。现报告如下。材料和方法一、实验动物健康雄性Ｗｉｓｔａｒ和ＳＤ大鼠（２００ｇ～３００ｇ）分别为供、受体，第三品系为Ｆ３１１大鼠。新西兰大白兔２．５ｋｇ～３．０ｋｇ。二、脾细胞悬液制备按本实验方法制成细胞浓度为１×１０８／ｍｌ，活力大于９４％。三、淋巴细胞悬液制备无菌取ＳＤ大鼠肠系膜淋巴结制成细胞悬液，采用Ｆｉｃｏｌｌ分离法制成淋巴细胞悬波（１×１０９／ｍｌ）。四、ＡＬＳ的制备及检测．用ＳＤ…     

T淋巴细胞疫苗诱导大鼠移植心存活时间延长的机制探讨

目的 探讨Ｔ淋巴细胞疫苗（ＴＣＶ）对大鼠移植心脏存活时间的影响及其机理。方法 取经供者抗原致敏的受者脾细胞（１／３脾切除）制备ＴＣＶ,然后将其回注于自身腹腔,观察大鼠异位移植心脏的存活时间。结果 接种ＴＣＶ可明显延长大鼠同种移植心的存活时间;注射ＴＣＶ后,Ｔ淋巴细胞增殖反应增强,Ｂ淋巴细胞增殖反应未受影响;混合淋巴细胞反应（ＭＬＲ）降低;脾细胞及ＴＣＶ表型分析显示ＣＤ８＾＋细胞增多;抗体介导的补体     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

目的 联合阻断OX40/OX40L和CD28/B7双协同信号途径和供体特异性脾细胞输注,诱导预存同种反应性记忆T淋巴细胞大鼠对同种心脏移植物的耐受.方法 建立大鼠预存同种反应性记忆T淋巴细胞的移植模型,采用免疫磁珠法分选CD8+CD44-记忆性T细胞;对过继转移供体特异性CD8+记忆性T细胞3 d的Lewis大鼠分别/合并输注AdCTLA4Ig、AdOX40Ig、供体脾细胞(DST),同时移植来至DA大鼠的心脏;48 h后取出心脏进行组织学分析、细胞因子表达分析,同时观察不同处理移植心脏存活时间.结果 AdCTLA4Ig+AdOX40Ig+DST组分别与AdCT-LA4Ig,AdOX40Ig和DST比较,心脏组织病理级别较低,白细胞介素(IL)-2及干扰素(IFN)-γ的表达水平也明显比其他组低很多,而心脏存活时间显著延长.结论 联合阻断OX40/OX40L和CD28/B7和供体特异性脾细胞输注能较好地诱导大鼠心脏移植物耐受.     

AG490延长移植心存活时间的探讨

目的 研究AG490在大鼠心脏移植中免疫抑制及延长移植心存活时间的作用,探讨AG490的作用机制.方法 供者为SD大鼠,受者为Wistar大鼠,建立大鼠心脏移植模型.将受者分为4组.对照组(14只):术后1～7 d经尾静脉注射生理盐水0.2 ml·kg-1·d-1;AG490组(14只):术后1～7 d经尾静脉注射AG490 20 mg·kg-1·d-1;CsA组(10只):术后1～7 d经尾静脉注射CsA20 mg·kg-1·d-1;AG490+CsA组(10只):术后1～7 d经尾静脉注射AG490和CsA各20mg·kg-1·d-1.术后各组分别取10只受者,观察移植心存活时间,并在术后1、4和7 d时,检测受者外周血中白细胞介素(IL)-2和IL-6的水平.术后第7天,处死对照组和AG490组受者(各4只)后,分别取移植心组织进行病理学检测.结果 AG490组受者术后移植心存活时间为(26.6±3.81)d,CsA组为(28.4±4.25)d,AG490+CsA组为(31.8±4.39)d,均较对照组的(8.4±0.84)d显著延长(P＜0.05).与对照组相比,AG490组术后外周血IL-2水平显著降低(P＜0.05),IL-6水平虽有所降低,但差异无统计学意义(P＞0.05),而AG490+CsA组IL-2和IL-6水平下降更为显著,与对照组相比,差异均有统计学意义(P＜0.05).AG490组移植心组织中淋巴细胞浸润程度较对照组明显减轻.结论 AG490具有免疫抑制作用,能延长移植物的存活时间,与CsA联合应用时效果更加明显.AG490的主要作用机制包括降低IL-2表达的水平,抑制移植心组织中淋巴细胞的浸润.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

协同信号途径阻断和供体脾细胞输注诱导预存同种记忆性T淋巴细胞大鼠心脏移植物的耐受

Objective To induce the tolerance to cardiac allograft by combined blockade of OX40/OX40L and CD28/B7 co-stimulatory signaling and donor specific spleen cell transfusion in rat mod-els of pre-existent allogenic reactive memory T lymphocytes. Methods Lewis rats that underwent 3 days of adoptive transfer of donor specific CD8+ memory T cells, separated by immunomagnetic bead separation kit, received either separate or combined transfusion of AdCTLA4Ig, AdOX40Ig, donor spleen cells (DST) and transplantations of hearts of DA rats at the same time. Cardiac allografts were taken out 48 h after transplantation for histological analysis and cytokines expression ,and survival time of cardiac allografts with different treatments was observed. Results Compared with AdCTLA4Ig, AdOX40Ig and DST groups, AdCTLA4Ig + AdOX4OIg + DST group showed lower cardiac pathological grade with much lower expression level of IL-2 and IFN-γ,and much longer heart survival time. Conclusion Combined blockade of OX40/ OX40L and CD28/B7 and donor specific spleen cell transfusion could induce rat cardiac graft tolerance.     

通过小鼠皮肤移植后心脏移植模型观察记忆性T淋巴细胞的变化和作用

目的 建立小鼠皮肤移植后心脏移植模型,观察记忆性T淋巴细胞的变化和作用,及其与皮肤移植后同种反应性记忆性T淋巴细胞过继转移模型的差异和机理.方法 应用显微外科技术及血管套管法,建立小鼠皮肤移植后心脏移植模型(实验组),并以心脏移植前输注或未输注记忆性T淋巴细胞的2个组作为对照(移植对照组和输注对照组),两对照组均未进行皮肤移植.心脏移植后每天触摸各组供心的搏动情况,观察各组移植心的存活情况及组织病理学改变,并对排斥反应进行分级;采用流式细胞仪检测各组受鼠体内CD3+ CD44high CD62L-记忆性T淋巴细胞表型的分布,应用体外混合淋巴细胞反应体系综合评价各组记忆性T淋巴细胞体外增殖程度.结果 皮肤移植前受鼠体内CD4+记忆性T淋巴细胞的比例为2.7％,移植后升至72.0％ (P＜0.05).移植对照组、输注对照组及实验组移植心脏的平均存活时间分别为7.33、4.83和3.5d,排斥反应评分分别为1B～2级、3A级和4级,体内CD3+ CD44highT淋巴细胞的比例分别为29％、55％和75％,CD3+ CD62L-T淋巴细胞群落中CD44荧光强度分别为215、274和380,以及MLR强度(吸光度值)分别为0.29、0.45和1.0,与两对照组相比,实验组移植心存活时间明显缩短、记忆性T淋巴细胞比例明显升高、MLR强度均明显增强及排斥反应程度更严重(P＜0.05或P＜0.01).结论 同种小鼠皮肤移植后心脏移植模型是一个更为贴近临床实际的动物模型,其同种抗原反应能力明显强于过继输注同种记忆性T淋巴细胞的小鼠心脏移植模型,有利于免疫记忆的相关研究.     

鼠纤维介素在小鼠心脏移植急性排斥反应中的表达及其意义

目的　探讨小鼠同种心脏移植急性排斥反应期间鼠纤维介素 (mfgl2 )在移植心脏组织中的表达及其与组织病理学改变的关系。方法　采用BALB/c小鼠到C5 7BL/ 6小鼠的颈部异位心脏移植作为同种排斥反应模型 ,以抗mgfl2多克隆抗体干预 ,并设同系小鼠心脏移植对照组。采集移植心组织标本作病理学检查 ,以免疫组织化学方法测定mfgl2在移植心脏组织细胞上的表达 ,并对mgfl2的表达进行半定量分析。结果　同系移植对照组移植心脏组织结构正常 ,未见mfgl2表达 ;同种移植组移植心脏组织出现进行性坏死 ,大量单个核细胞浸润 ,并伴有mfgl2的表达 ,且血管内皮细胞表达mfgl2 ;抗mfgl2抗体干预组移植心脏组织损伤较轻 ,移植物的存活时间延长 (P <0 .0 1) ,巨噬细胞、淋巴细胞的浸润和mfgl2表达量也显著减少。结论　mfgl2表达水平与排斥反应所致移植心脏病理损害程度相关 ;抗mfgl2抗体干预能显著减少移植心脏巨噬细胞、淋巴细胞的浸润 ,明显延长移植心脏存活时间。     

脾切除对心脏移植大鼠外周血淋巴细胞凋亡及调节性T淋巴细胞的影响

目的 探讨脾切除对同种异体心脏移植大鼠外周血淋巴细胞凋亡及调节性T淋巴细胞的影响.方法 以Wistar大鼠为供者、SD大鼠为受者,进行腹部异位心脏移植,同时切除受者的脾脏(心脏移植切脾组),并以不切脾者为对照(心脏移植对照组),另设不行任何处理的对照组和单纯切脾的单纯切脾组.术后第1、3、5、7天.取各组受者的移植心脏和外周血,观察移植心脏的组织学变化和细胞超微结构改变情况,以流式细胞仪检测外周血淋巴细胞的凋亡率及CD4+ CD25+ T淋巴细胞的变化,逆转录聚合酶链反应检测CD4+ CD25+ T淋巴细胞上Foxp3 mRNA的表达情况,记录移植心脏的存活时间.结果 心脏移植对照组移植心脏存活时间为(7.47±2.24)d,心脏移植切脾组移植心脏存活时间为(17.63±4.54)d,二者间的差异有统计学意义(P<0.05).心脏移植对照组的移植心脏肿胀,质硬,色暗,间质水肿、出血,弥漫性炎症细胞浸润,大量心肌细胞坏死、溶解,横纹不清;心脏移植切脾组的移植心脏质软,色红,局部灰白,外膜下以及细胞间局灶性水肿,炎症细胞浸润,心肌细胞结构完整,横纹清晰;心脏移植切脾组的细胞超微结构改变轻于心脏移植对照组.心脏移植切脾组术后第5天和第7天的淋巴细胞凋亡率分别为(7.62±2.15)%和(9.41±3.82)%,明显高于心脏移植对照组(P<0.05,P<0.05).心脏移植切脾组术后第3、5、7天时的CD4+ CD25+ T淋巴细胞明显多于心脏移植对照组(P<0.01,P<0.01,P<0.01),其Foxp3 mRNA的表达也较心脏移植对照组明显上调.结论 脾切除使心脏移植大鼠外周血淋巴细胞凋亡率增加,调节性T淋巴细胞增多,其Foxp3 mRNA表达上调,这些变化与移植心脏病理改变呈负相关.     

供者骨髓基质干细胞输注延长大鼠移植心存活时间的探讨

目的 探讨供者骨髓基质干细胞(MSC)输注在大鼠同种心脏移植术后的免疫调节及延长移植心存活时间的作用.方法 供者为近交系Wistar大鼠,受者为Fisher 344大鼠.处死供者后抽取其股骨和胫骨中骨髓,分离和培养MSC.通过混合淋巴细胞试验观察不同密度的MSC对异源性T淋巴细胞增殖反应的抑制作用.建立大鼠异位心脏移植模型,根据处理方式的不同,将受者分为MSC输注组和对照组,每组8只.Msc输注组:将含有2×106个MSC的林格氏液分别于术前1周、术中及术后连续3 d经尾静脉注入受者体内;对照组:用与MSC输注组相同的方法在相同时间点注入不含MSC的林格氏液.术后第5天,采用实时逆转录聚合酶链反应检测移植心组织中细胞因子的表达情况.结果 供者MSC可明显抑制异源性T淋巴细胞的增殖反应,且MsC密度越高抑制作用越强.MSC输注组Th1类细胞因子白细胞介素(IL)-1β和γ干扰素的表达要显著低于对照组;MSC输注组Th2类细胞冈子IL-4和IL-10呈高表达,而对照组基本不表达.MSC输注组移植心平均存活时间为(12.4±5.3)d,与对照组的(6.4±2.0)d比较,差异有统计学意义(P＜0.01).结论 输注供者MSC可通过改变Th1/Th2类细胞因子的平衡向Th2偏移诱导受者产生免疫调节作用,从而延长移植心存活时间.     

改良法分离小鼠移植心内浸润淋巴细胞

目的 建立简便而高效的分离小鼠心脏移植物内浸润淋巴细胞的方法.方法 移植物剪碎后用Ⅱ型胶原酶(250 U/ml,30～40 min)消化,Ficoll密度梯度离心(800 g/min,20 min)分离单个核细胞,荧光标记抗体行表面抗原或胞内细胞因子染色后用流式细胞仪分析其亚群.结果 移植物分离的单个核细胞数量稳定在1 × 106个以上,淋巴细胞的比例为(31.9±2.3)%,活性(95.1±2.1)%,流式细胞仪能同时检测到荧光标记的淋巴细胞表面和胞内的抗原.结论 本法单用胶原酶消化移植心,减少了对心脏移植物内浸润淋巴细胞的损伤,获得的细胞可进一步用于流式表型分析.