雷帕霉素延长大鼠同种异体肾上腺移植物存活的作用

目的　观察鼠同种异体肾上腺移植的效果及雷帕霉素 (Rapa)的作用。方法　采用显微外科技术进行 18只大鼠同种异体带蒂肾上腺移植。通过假手术组 ,双侧肾上腺切除组、纯系鼠肾上腺移植、同种异体肾上腺移植对照组及雷帕霉素治疗组在血、尿醛固酮值 ,存活期及病理进行比较。结果　鼠同种异体肾上腺移植存在显著的排斥现象 ,移植鼠术后平均存活 10 .7d ,而术后口服雷帕霉素 0 .4mg .kg-1·d-12周的移植鼠平均存活 2 3 .5d。结论　雷帕霉素能延长鼠同种异体肾上腺移植的存活期 ,但雷帕霉素在鼠同种异体肾上腺移植中应用最佳时间及副作用有待进一步研究。     

羟喜树碱和环孢素A对异基因大鼠心脏移植物急性排斥反应的协同抑制作用

目的 探讨羟喜树碱（OPT）和环孢素A（CsA）对异基因心脏移植物急性排斥反应的协同抑制作用。方法 以纯系SD大鼠为供者,纯系Wistar大鼠为受者,行异体颈部异位心脏移植。40只受者大鼠心脏移植后随机分成4组。A组接受安慰剂。B组接受 OPT1.0mg·kg-1·d-1,腹腔注射。E组接受CsA　10mg·kg-1·d-1,导管灌胃。F组联合应用OPT和CsA,方法剂量同B、E组。结果A组平均存活期为（6.05±0.76）d,B组为（11.45±1.99）d,E组为（14.45±4.85）d,F组有5只大鼠平均存活期为（45.00±19.43）d,另5只大鼠在移植60d后停用OPT和 CsA,存活期超过730 d,并经试验证明形成特异性免疫耐受。结论OPT和CsA的联合应用显著降低异基因大鼠心脏移植物急性排斥反应,明显延长心脏移植存活期。     

Fas配体阳性睾丸细胞和环孢素A对移植胰岛细胞存活起协同保护作用

目的　探讨Fas配体 (FasL)阳性睾丸细胞与胰岛细胞共移植后联用环孢素A(CsA)对移植胰岛细胞存活的协同保护作用。　方法　将同种大鼠胰岛细胞与睾丸Sertoli细胞同侧或异侧共移植于 4 1只糖尿病SD大鼠受体肾包膜下。实验大鼠分 7组 ,术后酌用CsA ,观察各组大鼠移植物存活情况。　结果　单纯胰岛细胞移植 (对照组 )后胰岛细胞的平均存活期为 (4 6± 1 1)d ,加用CsA存活期明显延长至 (2 1 8± 4 7)d(P <0 0 1)。与 1× 10 7个睾丸细胞同侧共移植的胰岛细胞平均存活期超过 (5 7 5± 4 0 )d ,但如移植前先封闭睾丸细胞表达的FasL后 ,移植的胰岛细胞平均存活期缩短为(5 8± 2 6 )d。胰岛细胞与 1× 10 5个睾丸细胞分别共移植于两侧肾包膜下 ,术后联用CsA ,胰岛细胞的平均存活期超过 (5 5 0± 6 5 )d ,与 1× 10 7个睾丸细胞同侧共移植的存活期相近 ,但比对照组或CsA组则显著延长 (P <0 0 1)。当胰岛细胞与 1× 10 6个睾丸细胞分别共移植且不用CsA时 ,胰岛细胞存活期平均仅为 (11 5± 3 1)d ,但仍较对照组延长 (P <0 0 5 )。　结论　表达FasL的睾丸细胞与CsA联用后可通过不同机制抑制胰岛细胞移植排斥反应而起到全身的协保护作用。     

小剂量环孢素A对大鼠同种异体肢体移植的免疫抑制作用

目的通过大鼠同种异体肢体移植模型,分析小剂量环孢素A(cyvlosporinA ,CsA)对大鼠同种异体肢体移植急性排斥反应的免疫抑制作用。方法采用雄性Wistar和SD大鼠为供、受体,以CsA为免疫抑制剂。对照组14只,将Wistar大鼠肢体移植至SD大鼠,术后不用药。实验组术后按用药剂量的不同分为2组( 2mg/kg组17只大鼠和CsA 6mg/kg组18只大鼠)。2组术后用药时间均为4周(每日1次共2周,然后每周2次共2周)。术后观察大鼠一般情况、移植肢体排斥反应及存活时间;用免疫荧光染色流式细胞仪检测各组手术前后T细胞亚群的变化。结果对照组移植肢体平均存活时间为[( 7.0 0±0 .78)d , x±s ,下同] ;CsA 2mg/kg组为( 3 7.18±0 .5 1)d ,CsA 6mg/kg组为( 3 3 .2 0±1.0 5 )d。术后第3天,对照组的CD4/CD8比值显著增高,移植肢体开始出现肿胀、皮肤红斑等表现。实验组的CD4/CD8较术前无明显变化。结论小剂量CsA能抑制大鼠同种异体肢体移植术后急性排斥反应的发生,并能延长移植肢体的存活时间。     

环孢素A与盐酸小檗碱联用预防大鼠心脏移植排斥反应

目的探讨环孢素A(CsA)与盐酸小檗碱联用对同种异基因大鼠心脏移植排斥反应的抑制效果。方法建立Wistar大鼠心脏到SD大鼠的异位(腹腔内)心脏移植模型,术后单独使用CsA、盐酸小檗碱以及联合使用CsA和盐酸小檗碱进行免疫抑制治疗,观察各组移植心脏的存活时间。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<0.01);CsA和盐酸小檗碱联用组移植心脏的存活时间较单用小剂量CsA组明显延长(P<0.01);单用盐酸小檗碱组与安慰剂组相比,移植心脏存活时间的差异无统计学意义(P>0.05)。结论CsA与盐酸小檗碱联用预防大鼠心脏移植后的排斥反应具有协同作用。     

FK506免疫抑制下异体肢体移植的长期存活

在实验性肢体移植中,人们使用了跨越不同免疫屏障的动物模型,以及多种不同的免疫抑制剂。在第1例动物肢体移植实验中采用间生态,之后是输血、抗淋巴细胞血清、6-琉基嘌呤、硝基咪硫嘌呤和类固醇,以及环孢素(CsA),它们显著延长了同种异体大鼠肢体移植的存活期,同时存活期具有浓度和生存状态依赖性。尽管如此,如何预防移植后的慢性排斥仍是个难题,药物明显的毒副作用(肝毒性、肾毒性)也使选择性复合组织移植难以被普遍接受,尤其在无生命威胁的情况下。另外,包含皮肤和肌肉的同种异体复合组织的确比实质器官具有更大的抗原性,要求更高的免疫抑制药物浓度。 Jones等对几种新药物,包括FK506、RS-61443和雷帕霉素,以及联合免疫抑制治疗在实验性同种异体复合组织移植中     

T细胞疫苗诱导大鼠同种异体肢体移植免疫耐受的实验研究

目的 探讨T细胞疫苗（TCV）诱导大鼠同种异体肢体移植特异性免疫耐受的作用。方法 制备受者Lewis大鼠针对供者DA大鼠的TCV，应用TCV，免疫正常Lewis大鼠共3次，每周1次。设TCV组和TCV未接种组，在接种前及接种后5d进行混合淋巴细胞培养（MLR）；设TCV组、CsA组和空白对照组，于接种后7d进行DA大鼠针对Lewis大鼠的同种异体肢体移植，在术后7d进行淋巴细胞毒检测，术后21d进行嵌合分析。结果 MLR显示，Lewis大鼠的脾细胞反应程度接种组显著低于未接种组（P〈0．01）；微量细胞毒测定显示，死亡细胞百分率在TCV组、CsA组、空白对照组3组比较性差异有统计学意义（P〈0．01）；嵌合分析显示经处理的Lewis大鼠脾脏，胸腺中检测出了DA大鼠源性的骨髓嵌合体。结论 T细胞疫苗可以抑制受者对供者的免疫应答；作为骨髓移植前预处理手段，T细胞疫苗接种后行吻合血管的骨髓移植成功地诱导出同种异体肢体移植嵌合耐受。     

供肝冷缺血时间延长诱发大鼠原位肝移植术后早期急性排斥反应的研究

目的 研究供肝冷缺血时间延长对大鼠原位肝移植术后早期急性排斥反应的影响.方法 选取30只健康纯系清洁级BN大鼠和30只Lewis大鼠.分别作为纯系移植和同种异体移植的供者,受者均为健康纯系清洁级BN大鼠60只,建立纯系和同种异体原位肝移植模型.根据纯系移植和同种异体移植供肝冷缺血时间的不同,将供、受者分为A、B、C和D组,每组15对.A组:供肝冷缺血1 h后进行纯系移植;B组:供肝冷缺血18 h后进行纯系移植;C组:供肝冷缺血1 h后进行同种异体移植;D组:供肝冷缺血18 h后进行同种异体移植.术后观察受者的2周存活率、移植肝组织病理学及肝功能的改变,检测受者主要组织相容性复合物(MHC)-Ⅱ类分子和核转录因子κB(NF-κB)的表达水平.结果 肝移植术后2周,A、B、C和D组的存活率分别为83.3%、66.7%、16.7%和0%,不管是纯系移植组还是同种异体移植组中供肝的冷保存时间越短,受者的存活率越高,且经肝功能检查发现冷保存时间短的受者移植肝功能恢复较好,移植肝组织病理学损伤和急性排斥反应也明显较轻.B组受者术后移植肝大量表达MHC-Ⅱ类分子,明显高于A组(P＜0.05);两同种异体移植组MHC-Ⅱ类分子的表达量较两纯系移植组增加明显,D组增加最多.A组几乎不表达NF-κB,而B组NF-κB的表达显著增加(P＜0.05);两同种异体移植组受者NF-κB的表达峰值提前.结论 冷缺血时间的延长可以诱导发生和加重大鼠原位肝移植术后早期急性排斥反应,降低术后2周存活率.     

FTY720诱导大鼠心脏移植物长期存活

目的 观察FTY720对大鼠同种异体心脏移植物存活时间的影响。方法 进行SD Wistar大鼠的腹部异位心脏移植，将受者随机分为对照组、甲泼尼龙(MP)组、环孢素A((SsA)组、FTY720组、FTY720与CsA二药联用组和FTY720、CsA及MP三药联用组，各组按分组要求分别于术前3d至术后14d通过灌胃给予FTY720和CsA，术前1d至术后2d腹腔注射给予MP，观察各组动物术后外周血淋巴细胞数量变化和移植物存活时间。结果 FTY720组、二药联用组和三药联用组的大鼠外周血淋巴细胞在给药后3h开始明显下降，停药后开始回升，至停药14d后恢复正常；移植心脏的存活时间，对照组平均为7．8d，CsA组为16．0d，MP组为27．6d，三药联用组为16．8d，而FTY720组和二药联用组分别超过了150d和124d。结论 FTY720可诱导同种异体大鼠心脏移植物长期存活。     

输注供鼠骨髓间充质干细胞延长肾移植大鼠的存活时间

目的 探讨在同种大鼠肾移植中输注供者骨髓间充质干细胞(MSC)对急性排斥反应的影响以及延长受鼠存活时间的作用.方法 取Wistar大鼠骨髓,分离和培养其MSC.以Wistar 大鼠为供者,Lewis大鼠为受者,建立同种大鼠肾移植模型.根据受鼠处理方式的不同,分为低剂量MSC组、高剂量MSC组、CsA组及对照组,低剂量MSC组和高剂量MSC组于移植前后分别多次输注1×106个和t×107个供者MSC,CsA组于术后2d开始腹腔内注入CsA 0.5 mg·kg-1 ·d-1,以腹腔内注射PBS作为对照.移植后,比较各组受鼠的存活时间,观察各组移植肾功能及移植肾组织病理学改变.结果 低剂量MSC组、高剂量MSC组、CsA组及对照组受鼠的存活时间分别为(21.7±7.2)d、(31.2±14.3)d、(34.9±15.7)d及(9.0±2.3)d;低剂量MSC组、高剂量MSC组和CsA组存活时间均明显长于对照组(P＜0.01),而低剂量MSC组存活时间明显短于高剂量MSC组和CsA组(P＜0.05).术后第4天,高剂量MSC组和CsA组移植肾组织形态和结构基本正常;对照组移植肾组织则表现出典型的急性排斥反应,出现广泛间质性浸润,肾小管炎症和片状坏死、出血,肾小球炎症浸润严重;而与对照组相比,低剂量MSC组急性排斥反应的病理表现则要明显减轻.结论 同种肾移植大鼠输注供者MSC后,可以达到有效的免疫调节作用,并且可明显延长大鼠的存活时间,呈MSC剂量依赖性.     

雷帕霉素对大鼠心脏移植的免疫抑制效果

为了探讨雷帕霉素对ＳＤ、Ｗｉｓｔａｒ大鼠心脏移植的免疫抑制效果，将大鼠分为３组，实验组给ＲＰＭ，对照且包括未治疗组及环孢素组。结果显示ＲＰＭ为一强效免疫抑制剂，治疗优于ＣｓＡ。ＲＰＭ具有强大的免疫抑制效果，可显著延长心脏移植物的存活时间。     

Immunosuppressive activity of FTY720, sphingosine 1-phosphate receptor agonist: I. Prevention of allograft rejection in rats and dogs by FTY720 and FTY720-phosphate

FTY720, a new class of immunomodulator, induces lymphopenia by sequestration of circulating lymphocytes into secondary lymphoid tissues. FTY720 at 0.1 to 1 mg/kg significantly prolonged the allograft survival in a dose-dependent manner and showed a marked synergistic effect in combination with cyclosporine (CsA) in rat skin and cardiac allograft models. In addition, the canine renal allograft survival was significantly prolonged by combination therapy with FTY720 at 0.03 to 1 mg/kg and CsA at 10 mg/kg as compared with monotherapy of FTY720 or CsA. By contrast, the combination therapy with CsA and azathioprine or CsA and mycophenolate mofetil resulted in only an additive effect in rat skin allograft. When FTY720 was administered to rats, FTY720 was metabolized by omega-oxidation of the octyl side chain, and beta-oxidation subsequently, or phosphorylated by sphingosine kinase. Omega- and beta-oxidized 4 metabolities of FTY720 at 10 mg/kg i.v. showed neither lymphopenia nor immunosuppressive activity in rat skin allograft. On the other hand, (S)-enantiomer of FTY720-phosphate at 0.1 and 1 mg/kg intravenously induced a marked lymphopenia and significantly prolonged the allograft survival in the rat allotransplantation. From these results, it is suggested the lymphopenia and the immunosuppression induced by FTY720 administration is due to the agonistic activity against SIP receptors of the active metabolite, (S)-FTY720-phosphate.     

Reversal of cyclosporine-induced mortality with a synthetic polymeric immunostimulant in a murine model of fecal peritonitis

We have been investigating the effects of a synthetic immunostimulative polymer known as copovithane (Cpv). This agent appears to enhance humoral immunity in untreated and cyclosporine-immunosuppressed mice and is nontoxic in rodents and man. The purpose of this study was to determine whether cyclosporine (CsA) is deleterious to survival in a murine cecal ligation, puncture, and excision (CLPE) model of fecal peritonitis, and--if so--whether this effect could be ameliorated by Cpv without interfering with skin allograft acceptance. Cpv significantly prolongs survival in the CLPE model; the optimal dose for this effect was found to be 100 mg/kg. CsA was found to have a significant and deleterious effect on survival at several dosage levels when administrated 48 and 24 hr before cecal ligation, and immediately before and 16 hr after cecal ligation. Using a dose of CsA sufficient for skin allograft acceptance and the same schedule of administration outlined above, Cpv 100 mg/kg was administered 48 hr prior to cecal ligation. Mice treated with CsA plus Cpv had significantly longer survival than mice treated with CsA alone; furthermore, the survival of CsA-plus-Cpv-treated animals was not significantly different from that of saline-treated controls. Acceptance and survival of H-2 incompatible skin allografts in mice treated with CsA were not affected by Cpv 100 mg/kg/week. We conclude that CsA-induced mortality in the CLPE model can be abrogated by Cpv without adversely affecting skin allograft survival. It may eventually be possible to reduce the incidence of septic complications in clinical allotransplantation by prophylactically administering Cpv to patients on CsA immunosuppression.     

The transfusion effect is influenced by the nature of MHC antigen presentation

We designed a study to determine whether the mode of presentation of major histocompatibility antigens is important for the ability of donor-specific blood transfusions to prolong organ allograft survival. Donor BN rat whole blood, isolated RBC, RBC ghosts, RBC membrane fragments, or whole blood lysates were administered to Lewis rat recipients 7 days before heterotopic allotransplantation of BN hearts. Only allogeneic whole blood or RBC significantly prolonged cardiac allograft survival in this histoincompatible donor-recipient pair. Whole blood lysates or RBC ghosts and membrane fragments transfused pretransplant did not prolong cardiac allograft survival when compared with syngeneic, transfused control rats. These results suggest that the immunosuppressive effects of donor-specific pretransplant blood transfusion may depend on a three-dimensional spatial relation between cells bearing donor major histocompatibility antigens and recipient responder cells responsible for the transfusion effect.     

氨基胍与环孢素A联用对大鼠心脏移植后急性排斥反应的影响

目的：探讨氨基胍与环孢素A联用对同种大鼠心脏移植后急性排斥反应的影响。方法：受体SD大鼠心脏移植后分为4组：（1）对照组：术后不作任何处理;（2）低剂量环孢素A（CsA)组;术后0－7d肌肉注射CsA2mg.kg^-1.d^-1;（3）氨基胍（AG）组：术后0－7d皮下注射AG600mg.kg^-1.d^-1;（4）低剂量CsA加AG组;术后0－7d肌肉注射CsA2mg.kg^-1.d^-1及皮下注射AG600mg.kg^-1.d^-1。术后4d测定急性排斥反应时移植心的诱生型一氧化氮合酶（iNOS)的表达及血清一氧化氮（NO）含量,并观察移植心存活时间。结果与低剂量环孢素A组相比较,低剂量环孢素A与氨基胍联用组不仅显著地抑制移植心iNOS表达与NO产生（P＜0．05）;而且显著地减轻急性排斥反应（P＜0．01）,延长了移植心存活时间（P＜0．05）。结论低剂量环孢素A与氨基胍联用,协同抑制急性排斥反应时移植心iNOS活性及NO产生;显著地延长移植物存活时间。     

Immunosuppressive activity of the Chinese medicinal plant Tripterygium wilfordii. I. Prolongation of rat cardiac and renal allograft survival by the PG27 extract and immunosuppressive synergy in combination therapy with cyclosporine

BACKGROUND: PG27 is an immunosuppressive fraction purified from an extract of a Chinese medicinal plant, Tripterygium wilfordii. We tested PG27 in rat cardiac and renal allotransplantation, and we examined the immunosuppressive interaction with cyclosporine (CsA). METHODS: Brown Norway (BN) rat heart or kidney allografts were transplanted into the abdomen of Lewis rats, which were treated by the intraperitoneal or oral route with PG27, CsA, or both. RESULTS: PG27 administered intraperitoneally to Lewis recipients for 16 days at 10-30 mg/kg/day significantly increased the median survival time of BN heart allografts from 7 to 18-22 days. Oral administration was effective, with cardiac allograft survival prolonged to > 100 days with 52 days of treatment. PG27 at 20-30 mg/kg/day significantly extended the median survival time of BN kidney allograft recipients from 9 to 36.5-77 days, and 30 mg/kg/day for 52 days extended survival beyond 200 days. PG27 combined with CsA significantly enhanced heart and kidney allograft survival, even at doses of CsA ineffective when administered alone. The addition of 5 or 10 mg/kg/day PG27 reduced by 50-75% the CsA dose needed for 100% kidney allograft survival. The combination index was less than 1.0, indicating synergy of PG27 with CsA in prolonging cardiac and renal allograft survival. Furthermore, the PG27/CsA combination exerted a positive influence on renal allograft function. PG490 (triptolide, a constituent of PG27) and PG490-88 (a semisynthetic derivative of PG490) suppressed rejection of cardiac and renal allografts. CONCLUSIONS: The PG27 herbal extract demonstrated immunosuppressive activity by prolonging heart and kidney allograft survival, displaying synergy in the immunosuppressive interaction with CsA, and improving renal allograft function in combination with CsA. PG490 and PG490-88 compounds were also effective.     

Effect of a New Immunosuppressive Agent,FTY720, on Survival of Islet Allografts

A newly developed immunosuppressant, FTY720, has a unique mechanism that is quite different from those of conventional immunosuppressants, and is presumed to be mediated through decreases in the number of peripheral lymphocytes, especially helper T cells. This study was performed to ascertain whether this innovative drug could prolong islet allograft survival. The donors were inbred Lewis rats and the recipients were ACI rats rendered hyperglycemic with intravenous streptozotocin. In the study group, FTY720 dissolved in distilled water was orally administered at a dose of 5 mg/kg to the recipient ACI rats 1 day before and on the day of grafting. In the control group, only distilled water was orally administered to the recipient ACI rats on the day before and the day of grafting. Two thousand islets were transplanted into the portal vein of the recipient rats in the study and control groups immediately after isolation. The graft survival time in the study group was significantly longer than that in the control group, indicating that FTY720 retains a potent effect on the prolongation of islet allograft survival. FTY720 could become a useful immunosuppressant for future clinical islet allotransplantation.     

Effect of fusidic acid on pancreatic islet allograft rejection

We examined in fully mismatched rats, the survival of pancreatic islet allografts in recipients treated with either fusidic acid (FA), an antistaphyllococcal antibiotic that has been shown to possess an immunosuppressive effect in vitro and in vivo, or cyclosporin-A (CsA). Islets were isolated by collagenase digestion, separated from acinar tissue by handpicking under a dissecting microscope and transplanted into the liver by portal vein injection of streptozotocin(STZ)-induced diabetic rats. The results indicated that while a temporary immunosuppression with CsA achieved an indefinite islet allograft survival, FA administered to recipients daily was not able to prevent islet allograft rejection across a major histocompatibility barrier. We conclude that despite the fact that fusidic acid has been claimed to act as an immunosuppressant drug in vitro with effects similar to those of CsA, unlike CsA, FA given either orally or by s.c. injection was not effective to prolong islet allograft survival in vivo.     

Failure to prolong pancreatic islet allograft survival in rats with donor-specific blood transfusions and immunosuppression

Abstract. The effects on pancreatic islet allograft survival of donor-specific blood transfusions (DST) in combination with pre-and posttransplant immunosuppression were studied. A total of 12 groups of rats ( n = 105) with chemically induced diabetes underwent islet allotransplantation. Multiple DST or third-party blood transfusions (TPT) were given prior to transplantation. Pretransplant immunosuppression consisted of azathioprine and prednisolone, and low-dose cyclosporin A was used for posttransplant immunosuppression. TPT, as well as separate or combined pre-and posttransplant immunosuppression without blood transfusions, did not prolong islet allograft survival. DST resulted in either primary nonfunction of the islet allografts or a markedly decreased islet allograft survival. These findings contrast with the beneficial effect of DST on whole-organ allograft survival in rats previously described by others.