共刺激阻断剂与雷帕霉素联合应用对移植肾存活的影响

目的:探讨共刺激阻断剂CTLA4Ig与雷帕霉素(Rapamycin,RPM)联合应用对大鼠移植肾存活的影响.方法:肾移植大鼠分为对照组、CTLA4Ig组、RPM组和CTLA4Ig RPM组,观察术后血肌酐和移植肾病理改变、移植肾存活时间.结果:与对照组相比,CTLA4Ig组、RPM组、CTLA4Ig RPM组移植肾存活时间均显著延长(P＜0.01),其中,CTLA4Ig RPM组移植肾存活时间最长(76.5±17.1d),术后15天、30天该组血肌酐浓度最低;术后30天,仍然存活的CTLA4Ig组移植肾显示较多淋巴细胞浸润,CTLA4Ig RPM组移植肾显示很少淋巴细胞浸润.结论:共刺激阻断剂与雷帕霉素联合应用可有效抑制排斥反应,显著延长大鼠移植肾存活时间.     

CD154胞外区基因局部修饰对大鼠移植肾存活的影响

目的观察CD154胞外区基因局部修饰大鼠供肾对延长移植肾存活的效能.方法以CD154基因重组腺病毒为载体,将CD154胞外区基因转入BN大鼠供肾,以Lewis大鼠为受体,行同种肾移植为转染组,并以未转染BN供肾移植给Lewis受体为对照组;观察移植肾存活时间和术后肾功能变化.结果转染组移植肾存活(28±7.3) d,较对照组移植肾存活时间(8.6±1.2)d明显延长;移植组术后血清肌酐较同期对照组明显为低.结论 CD154基因局部转染可明显延长移植肾存活时间.     

抗ICOS单抗与小剂量环孢素A联用对大鼠心脏移植慢性排斥反应的影响

目的：探讨抗可诱导共刺激分子（inducible costimilator,ICOS）单克隆抗体与小剂量环孢素A（cyclosporine,CsA）联用对同种异基因大鼠心脏移植后移植物生存质量和慢性排斥反应的影响。方法：建立大鼠腹腔异位心脏移植模型,术后随机分为同基因移植对照组和同种异基因移植试验组,其中试验组根据术后处理不同又分为安慰剂组、常规剂量CsA组、抗-ICOS-单抗组、小剂量CsA组和抗-ICOS-单抗联合小剂量CsA组,观察各组大鼠移植心脏的存活时间及移植心组织病理学变化,对移植心长期存活（〉100d）的受体,应用流式细胞仪检测其外周血CD4^＋CD25^＋调节性T细胞。结果：单独使用不同剂量CsA者移植心脏的存活时间较安慰剂组明显延长（P〈0.05）;抗-ICOS-单抗联用小剂量CsA组移植心脏的存活时间较单用小剂量CsA组明显延长（P〈0.05）;与安慰剂组相比,单用抗-ICOS-单抗组移植心脏存活时间差异无统计学意义（P〉0.05）。移植心长期存活受体中,与常规剂量CsA组比较,联合治疗组移植心慢性排斥损伤明显减轻,而外周血CD4^＋CD25^＋调节性T细胞比例有显著升高（P〈0.05）。结论：抗ICOS单抗加小剂量CsA联合免疫治疗,可有效延长大鼠移植心存活时间和减轻慢性排斥反应,CD4^＋CD25^＋调节性T细胞的高水平表达与诱导移植耐受、抑制慢性排斥反应有重要关联。     

不同处理对大鼠移植心脏存活时间的影响

目的 观察供体脾细胞(SPC)与环磷酰胺(CP)联合预处理对大鼠移植心脏存活时间的影响.方法 应用大鼠颈部心脏移植模型,实验分为对照组(未干预组)、CsA组、SPC CP组.CsA组心脏移植后予环孢霉素A(CsA)10 mg/kg,SPC CP组采用供体SPC和CP预处理移植受体,然后行大鼠颈部心脏移植术.观察移植心脏存活情况及组织病理变化.结果 对照组、CsA组、SPC CP组移植心脏存活时间分别为(7.2±2.4)、(15.8±4.3)、(30.4±10.7)天;组织病理检查显示,经SPC PC预处理的移植心脏排斥反应明显减弱.结论 SPC和CP预处理可以诱导受体对移植物的免疫耐受,延长移植心脏的存活期,效果优于CsA.     

雷帕霉素对肾缺血再灌注大鼠肾功和肾组织超微结构的影响

目的:探讨雷帕霉素(Rapamycin,RPM)在大鼠肾缺血再灌注损伤时肾功能和肾组织超微结构改变中的作用.方法:30只雄性Wistar大鼠随机分为3组:假手术组(切除右肾,分离左肾动脉,不阻断血流,灌胃给等量生理盐水)、手术组(切除右肾,分离左肾动脉,阻断血流,缺血前灌胃给等量生理盐水)、药物组(切除右肾,分离左肾动脉,阻断血流,缺血前灌胃给RPM(4mg/(kg·d)×3d,最后一次术前2h灌胃).大鼠急性缺血性肾损伤模型为切除右肾,分离左肾动脉,阻断血流45min,再灌注24h,观察肾功能和肾组织超微结构改变.结果:肾缺血再灌注组血肌酐(168±37)μmol/L、血尿素氮(22±6 mmol/L,雷帕霉素组血肌酐(113±17)μmol/L、血尿素氮(13.8±2.3)mmol/L;2组比较差异有统计学意义(P<0.05).电镜:假手术组肾小管上皮细胞超微结构正常;手术组肾小管管腔被大量坏死脱落细胞碎片堵塞,上皮细胞胞浆内脂滴堆积,细胞有凋亡迹象;药物组肾小管超微结构损害轻微.结论:雷帕霉素对大鼠肾缺血再灌注损伤时肾功能和肾组织超微结构有明显保护作用.     

PDL1修饰树突状细胞对大鼠同种异体肾移植存活的影响

目的观察激活负性共刺激信号PD1-PDL1对延长移植肾存活的效能。方法以PDL1Ig基因重组腺病毒为载体,将PDL1Ig基因转入BN大鼠树突状细胞(DC)细胞同时输注受体,以Lewis大鼠为受体,行同种肾移植为转染组,并以未转染DC输注为对照组;观察移植肾存活时间和术后肾功能变化。结果转染组移植肾存活(41±7.6)d,较对照组移植肾存活时间(8.6±1.2)d明显延长;移植组术后血清肌酐较同期对照组明显为低。结论PDL1基因修饰的DC可以明显延长移植肾存活时间。     

新型合成肽延长大鼠移植肾存活时间的实验研究

目的探讨一种新型合成肽对淋巴细胞免疫功能及大鼠移植肾脏存活时间的影响。方法采用人工标准固相合成法新型肽,3H-TdR掺入法观察其在体外对人外周单核细胞增殖反应的影响,建立原位大鼠异基因肾移植模型32例,设合成肽加CsA治疗组、合成肽治疗组、CsA治疗组以及对照组4组,每组8例,观察大鼠移植肾脏存活时间及移植肾功能。结果合成肽可显著抑制淋巴细胞转化及MLR的增殖反应;围手术期应用合成肽结合小剂量CsA,能显著延长同种大鼠移植肾脏的存活时间(54d),并维持较低的血清肌酐水平〔(47.40±11.2)mmol/L〕。结论合成肽能显著抑制人外周单核细胞经丝裂原或同种抗原刺激引起的增殖反应,围手术期应用新型合成肽结合小剂量CsA,能显著延长异基因移植物存活时间及功能,表明合成肽可能成为一种新型的器官移植免疫调节药物。     

激活T细胞负性共刺激信号对大鼠同种异体肾移植存活的影响

目的观察激活负性共刺激信号程序性死亡因子及配体(PD1-PDL1)对延长移植肾存活的效能。方法以PDL1Ig基因重组腺病毒为载体,将PDL1Ig基因转入BN大鼠供肾,以Lewis大鼠为受体,行同种肾移植为转染组,并以未转染BN供肾移植给Lewis受体为对照组;观察移植肾存活时间和术后肾功能变化。结果转染组移植肾存活(36±7.3)d,较对照组移植肾存活时间(8.6±1.2)d明显延长;移植组术后血清肌酐较同期对照组明显为低。结论PDL1基因局部转染可明显延长移植肾存活时间。     

FasL基因转染诱导大鼠肾移植免疫耐受的实验研究

目的:探讨Fas配体(Fas ligand,FasL)在大鼠肾移植免疫耐受中的作用及意义.方法:首先建立大鼠同种肾移植模型.实验组供肾移植前用1ml含重组腺病毒Ad-FasL的肾脏冷保存液经肾动脉灌注;建立同种大鼠肾移植模型.分别观察电镜、免疫组织化学法、肾功能测定、生存期,比较各组间的不同.结果:Ad-FasL治疗组肾移植受体大鼠平均存活天数为31.3 d,与对照组的平均9.1 d存活时间比较,平均存活天数增加了22 d,差异显著.Ad-FasL治疗组的移植肾功能基本保持稳定,对照组在术后5 d切除对侧肾脏后血清肌酐浓度迅速上升.结论:腺病毒载体可成功介导FasL基因对大鼠肾脏的转染,并对移植肾起免疫保护作用、延长移植肾的存活时间.     

聚乙二醇对同种小鼠胰岛移植后抗排斥治疗的影响

目的 探讨聚乙二醇(PEG)包埋小鼠胰岛细胞对同种小鼠胰岛移植物存活的影响,以及其与抗排斥药雷帕霉素共同应用对胰岛移植后抗排斥治疗的影响.方法 选取C57BL/16雄性小鼠皮下预血管化糖尿病模型,行BALB/c小鼠胰岛移植.随机分为6组:A组:正常胰岛移植组.B组:PEG包埋胰岛移植组.C组:正常胰岛移植+雷帕霉素1.5 mg·kg~(-1)·d~(-1)治疗组.D组:PEG包埋胰岛移植+雷帕霉素1.5 mg·kg~(-1)·d~(-1)治疗组.E组:正常胰岛移植+雷帕霉素3 mg·kg~(-1)·d~(-1)治疗组.F组:PEG包埋胰岛移植+雷帕霉素3 mg·kg~(-1)·d~(-1)治疗组.观察小鼠血糖变化,病理检测移植物存活情况.结果 B组胰岛存活时间(25.5±5.9)d较A组(14.7±2.6)d明显延长(P<0.01),C组胰岛存活时间(35.0±3.1)d,D、E、F组胰岛于移植后6周均存活.HE染色下B组较A组炎症反应轻,胰岛结构完整.免疫组化染色显示B组有散在染色增强细胞团而A组未见.结论 PEG包埋胰岛细胞可显著延长移植物的存活时间,并能减少免疫抑制剂雷帕霉素的用量.     

雷帕霉素抑制大鼠高危角膜移植免疫排斥反应的实验研究

目的：研究雷帕霉素对大鼠高危角膜移植免疫排斥反应的抑制作用。方法：建立大鼠高危穿透性角膜移植动物模型，以角膜新生血管化sD大鼠为受体。Wistar大鼠为供体，分别腹腔注射0．5％羧甲基纤维素（对照组）、雷帕霉素、环孢霉素A、雷帕霉素＋环孢霉素A，共12d。用裂隙灯显微镜对角膜植片进行临床观察，记录角膜植片存活时间，并进行组织学检查。结果：用药组角膜植片的存活时间显著增加（P〈0．01）；组织学发现联合用药组的角膜炎性细胞浸润、新生血管形成度水肿程度均明显好于其他各组。结论：雷帕霉素能显著延长角膜植片的存活时间，对大鼠高危穿透性角膜移植排斥反应具有抑削作用，与环孢霉素A联合用药具有协同作用，优于各单药物治疗组。     

Rapamycin instead of mycophenolate mofetil or azathioprine in treatment of post-renal transplantation urothelial carcinoma

Background Malignant tumor is the most common complication occurred in transplant recipients. It is widely recognized that immunosuppressive treatments increase the risk of cancer in transplant recipients. The efficacy and safety of rapamycin （RPM） in combination with low-dose calcineurin inhibitor （CNI） in treating 15 renal allograft recipients which developed urothelial carcinoma were observed.
Methods Immunosuppressive regimen in all recipients was altered with rapamycin to replace mycophenolate mofetil （MMF） or azathioprine （Aza）. The initial loading dosage was 2 mg/d, and the next dosage was 1 mg/d. The dosage of rapamycin was carefully adjusted according to the blood drug level and concentration of the drug was maintained at 4-6 μg/L. In all the 15 patients, the calcineurin inhibitor was reduced down to one third of the original dosage after the rapamycin blood concentration became stable. Surgical treatment and intravesical instillation chemotherapy were carried out in all patients. Recurrence of the tumor was monitored throughout the study. Post-transplant renal function and side effects were also closely monitored.
Results Among the 15 patients, 9 had no tumor recurrence in 2 years, 2 had tumor recurrences twice, and 4 had once. There was no acute rejection observed during RPM treatment. Post-transplant renal function in 11 patients was improved with a decreased creatinine level. Hyperlipoidemia and thrombocytopenia were the most frequent adverse events which responded well to corresponding treatments.
Conclusion Among the renal allograft recipients with urothelial carcinoma, combination of rapamycin and low dose calcineurin inhibitor treatment is effective and safe.     

Effects of combined immune therapy on survival and Th1/Th2 cytokine balance in rat orthotopic liver transplantation

Background The induction of immune tolerance and suppression of allograft rejection has become the focus in the study of liver transplantation. The effect of immune therapy with anti-CD40L mAb alone or in combination with cyclosporine A (CsA) on the recipient survival and Th1/Th2 cytokine profile was studied to elucidate its immunological mechanism and role in rat orthotopic liver transplantation. Methods The model of rat orthotopic liver transplantation was established by modified Kamada’s technique. Recipients were divided into group A (control group): SD→SD; group B (group of rejection): SD→Wistar without any treatment; group C: SD→Wistar with CsA monotherapy from day 1 to day 5; and group D: SD→Wistar with CsA from day 1 to day 5 and anti-CD40L mAb on day 0 and day 2. The survival of the recipients in all groups was observed and ELISA technique was used to detect the level of cytokines in peripheral blood on post-transplant day 7. Results The survival period of recipients in groups A (&gt;60 days) and D (&gt;60 days) was significantly longer than that in group B (13.8±2.4 days). The serum levels of interleukin 2 (IL-2) and interferon γ in group B were significantly higher than those in other groups; the level of tumor necrosis factor α was higher but not statistically significant. In contrast, the serum levels of IL-4 and IL-10 in group D were elevated more significantly than those in group B (P&lt;0.05). Conclusions Combined immune therapy can prolong the survival of allografts. Increased expression of Th2 cytokines, which is closely related to the induction of tolerance and suppression of rejection, is beneficial to the long-term survival of recipients and allografts.     

供体脾灌注对高度致敏肾移植受者嵌合体形成的影响

目的探讨供体脾灌注对高度致敏肾移植受者稳定期嵌合体形成及移植肾功能的影响。方法对16例高度致敏患者进行配对分组,实验组肾移植术中先予供体脾灌注40min,前瞻性观察脾灌注对患者术后6个月内嵌合体形成、移植肾排斥反应发生及移植肾功能的变化。结果脾灌注后受者外周血中供者来源的有核细胞数量显著增加,受者形成稳定嵌合体的时间较早,例数比对照组更多;肾移植术后脾灌注组排斥反应发生时间较对照组延迟,排斥反应严重程度明显较对照组轻微;术后6个月时,脾灌注组患者血肌酐值低于对照组。结论供体脾灌注可以显著提高高敏肾移植受者外周血中供者来源的有核细胞数量,减轻排斥反应强度,从而促进受者嵌合体的形成,有利于改善稳定期移植肾功能。     

大剂量ARB对肾移植后蛋白尿治疗作用的临床研究

目的观察大剂量ARB对肾移植后蛋白尿的疗效及安全性。方法83例肾移植后蛋白尿患者,分为治疗组50例(采用2~4倍剂量ARB治疗)和对照组33例(不使用ARB治疗),观察不同治疗时间两组患者24h尿蛋白、血肌酐、血生化、血压、远期人/肾生存率及药物不良反应。结果治疗组24h尿蛋白排出显著低于对照组;治疗组血肌酐倍增时间、进展到移植肾功能衰竭或死亡的时间显著高于对照组〔(34.42±12.56)个月与(17.51±10.26)个月,(38.12±22.51)个月与(24.25±13.56)个月〕;治疗组出现2例血钾升高,4例低血压。结论大剂量ARB可有效控制肾移植后蛋白尿,提高移植患者人/肾远期存活率,使用安全有效。     

Strong additive effect of calcitriol and cyclosporine A on lymphocyte proliferation in vitro and rat liver allotransplantations in vivo

Background Vitamin D3 and its metabolites have been found to exert immunosuppressive effects both in vivo and in vitro. We investigated the synergistic effect of calcitriol and cyclosporine A (CsA) on lymphocyte proliferation in vitro and graft rejection following rat liver allotransplantations in vivo.Methods Alloantigen driven, human peripheral mononuclear cells’ proliferation and cytokine production capacity were tested in the presence or absence of various concentrations of calcitriol or CsA. In vivo, liver allografts were transplanted in a high responder strain combination (SD to Wistar) rats and combination of subtherapeutical dose of CsA and calcitriol was administered in recipients, whereas the control recipients received single or no immunosuppressant. Proliferation of splenocyte from recipient was tested with mixed lymphocyte reaction. Serum interleukin-2 (IL-2) and interferon gamma (IFN-γ) concentrations were measured with enzyme linked immunosorbent assay. Results Combined medication of 10(-9) mol/L calcitriol and 100 ng/ml CsA inhibited human peripheral mononuclear cells’ proliferation to alloantigen and the production of IL-2 and IFN-γ but promoted that of IL-4 and IL-10. Similarly, combination of 250 ng·kg(-1)·d(-1) calcitriol and 1.0 mg·kg(-1)·d(-1) CsA showed an additive effect in liver transplant model. It restrained splenocyte proliferation to alloantigen from donor and significantly reduced serum concentration of IL-2 and IFN-γ in recipients. Consequently, allograft rejection in combined medication group was minor (median William’s grade was 1.0 vs 3.0 in combined medication group and in the control group, P&lt;0.05) and the recipients’ survival was evidently prolonged [(93.7±5.8) days vs (12.6±1.4) days in combined medication group and in the control group, P&lt;0.01].Conclusion A combination of calcitriol and CsA has an additive effect on limiting lymphocyte proliferation and prolonging liver graft survival. With its additional immunomodulating property, calcitriol is a potent immunosuppressant that can extend the therapeutic window of classical immunomodulators in prevention and treatment of liver graft rejection.     

肾移植受者血清IL-10测定的临床意义

目的观察肾移植受者术后肾功能稳定、发生排异反应、感染、慢性移植肾肾病(CAN)时血清白细胞介素-10(IL-10)的水平;检测肾功能稳定者肾移植术后不同时间、环孢菌素A(CsA)剂量、排异次数时血清IL-10水平,以探讨IL-10在肾移植受者门诊随访中的意义。方法采用双抗体夹心酶联免疫吸附法,随机对127例门诊肾移植术后随访者血清IL-10水平进行检测。以20例健康志愿者作为对照组。结果肾功能稳定组患者的血清IL-10水平明显高于术后排异组、CAN组和对照组,相比较均有显著统计学意义(P<0.05);肾功能稳定组中CsA用量每千克体质量2 mg组的血清IL-10水平明显高于4 mg组,相比有统计学意义(P<0.05);该组中移植术后无排异者的血清IL-10水平明显高于1次排异和多次排异者,相比均有统计学意义(P<0.05)。结论血清IL-10水平可部分反映移植患者的免疫状态,对肾移植受者门诊随访有一定的参考价值。     

小剂量静脉滴注人免疫球蛋白在群体反应抗体阳性肾移植患者中的临床应用分析

目的  探讨群体反应抗体（PRA）阳性肾移植受者应用小剂量人免疫球蛋白（IVIG）后,PRA的下降程度以及移植肾功能的恢复效果。方法  ①观察组：A组术前高PRA肾移植患者4例;B组术后出现移植肾功能延迟恢复,监测出PRA升高患者7例;C组术后远期出现血肌酐升高,监测PRA升高同时病理穿刺证实为体液性排斥反应患者8例;②对照组：回顾性、随机挑选既往与A、B、C组同样表现的患者。治疗方案：A、B组血浆置换（PP）联合小剂量IVIG持续静脉滴注0.1 g/（kg·d）;C组内固醇激素冲击联合小剂量IVIG持续静脉滴注0.1 g/（kg·d）;对照组治疗方案中无IVIG的应用。比较观察组与对照组PRA下降程度和患者的预后。结果  经系统性、阶段性治疗后,A、B、C组中PRA均有不同程度降低,与对照组比较,A组肾移植术后排斥反应发生几率、B组移植肾功能恢复时间、C组移植肾血肌酐下降程度高于对照组,差异有统计学意义。结论  与传统大剂量IVIG比较,小剂量IVIG也能有效降低肾移植受者PRA水平,其对高PRA水平患者的治疗具有积极意义,且大大降低医疗成本。

     

尿激酶对环孢素A慢性肾病大鼠肾间质炎症的干预作用

目的 探讨外源性尿激酶型纤溶酶原激活物(uPA)对环孢素A(CsA)致慢性肾病大鼠肾间质炎症的干预作用.方法 将雄性SD大鼠低盐饮食饲养,经口服灌胃CsA(25 mg/kg·d)4周,制作慢性CsA肾病模型,分别给予小剂量uPA持续干预和大剂量uPA间断干预.4周末,观察大鼠血肌酐(Scr)、尿素氮(BuN)及24h尿蛋白的变化,Masson染色观察肾组织纤维蛋白沉积,免疫组化检测肾间质ED-1阳性巨噬细胞浸润数量、肾组织uPA及转化生长因子b1(TGF-?1)的表达.结果 大鼠经CsA灌胃后,Scr、BuN升高,24 h尿蛋白增加,肾间质纤维蛋白沉积增多,巨噬细胞浸润数量增加,肾局部uPA含量下降以及肾组织TGF-?1表达上调,表明大鼠肾间质炎症模型建立成功.小剂量uPA持续给药血Scr、BuN、24h尿蛋白水平下降(P<0.01或P<0.05),肾间质纤维蛋白沉积和炎性细胞浸润减轻(P<0.05),uPA表达增加(P<0.05),肾组织TGF-?1表达降低(P<0.05).大剂量uPA间断干预生化指标无显著性差异,局部uPA轻度升高,但对减轻肾小管间质纤维化和下调TGF-?1表达无显著性差异(P0.05).结论 小剂量uPA持续干预能有效减轻CsA慢性肾病大鼠肾间质纤维蛋白沉积、炎性细胞浸润及下调肾组织TGF-?1表达,从而减轻肾脏炎症反应和纤维化,为其临床应用提供了基础.     

青藤碱对肾移植大鼠IL-2的影响

目的 :研究青藤碱 (sinomenine,SIN)对肾移植大鼠存活时间及IL - 2的影响 ,探讨SIN免疫抑制作用的可能机制。方法 :实验分四个组 ,采用改良式大鼠肾移植术行Wistar→SD大鼠单肾移植 ,观测术后受体鼠的存活时间 ;ELISA法检测受体鼠外周血IL - 2水平。结果 :对照组受体鼠均在术后第 9天内死亡 ,平均存活时间 7.4± 0 .7d ,SIN组存活时间为 9.1± 1.0d ,而与低剂量CsA(2 .5mg/kg·d ,ip)联用后明显延长至 18d以上。SIN组受体鼠外周血IL - 2水平低于对照组 ,而SIN +CsA组受体鼠IL - 2水平明显低于对照组。结论 :SIN对大鼠肾移植的急性排斥反应具有一定的抑制作用 ,并与低剂量CsA产生显著的协同效应。SIN免疫抑制作用机制可能是抑制Th1细胞产生IL - 2 ,与CsA的作用机制相似。