去脑垂体大鼠诱导免疫耐受性的实验研究

在立体定向仪经耳去除近交系SD大鼠脑垂体基础上,用不同剂量近交新生的Wistar大鼠制成活化脾细胞和环磷酰胺进行诱导免疫耐受性的研究。结果:活化脾细胞每周5×10~7/0.5ml或5×10~7/0.5ml联合环磷酰胺20mg/（kg·d）持续2周后,去脑垂体大鼠同种异体皮肤移植平均存活时间分别为>33.5d和>47.8d。表明利用供体活化脾细胞或联合环磷酰胺能成功地诱导出受体对此供体的特异性免疫耐受。     

TLSFJM抑制大鼠心脏移植排斥反应的作用

目的　在同种异体大鼠异位心脏移植模型中 ,探讨TLSFJM对急性移植排斥反应的抑制作用及机制。方法　以F344大鼠作为心脏移植受体 ,以LOU/CN大鼠作为心脏移植供体 ,建立大鼠异位心脏移植模型。受体大鼠分为 3组 ,于移植前后分别施以RPMI16 4 0、CsA或TLSFJM。每天观察移植心脏跳动情况 ,并于停跳当天或之前解剖观察。分别取供体大鼠脾细胞作为刺激细胞 ,取受体大鼠脾细胞作为反应细胞 ,进行单向混合淋巴细胞反应。结果　TLSFJM可明显延长大鼠异位移植心脏的存活时间 :RPMI16 4 0对照组移植心脏的存活期全部为 6d ,TLSFJM治疗组最长均可存活 2 7d ,高剂量 (15mg/kg·d)CsA治疗组存活期超过 2 7d。TLSFJM治疗组大鼠脾细胞增殖的cpm值均低于对照组。结论　TLSFJM具有良好的抗急性移植排斥反应作用。TLSFJM对同种异体抗原诱导T细胞增殖的明显抑制 ,可能是其发挥免疫抑制作用的机制之一     

胸腺应急修饰诱导大鼠同种心脏移植免疫耐受

目的：与以往术前21天进行胸腺修饰常规方法不同，在手术当天进行胸腺内注射抗原，诱导大鼠同种心脏移植免疫耐受。方法：在大鼠腹腔异位心脏移植模型中，实验组在手术当天将供体SD大鼠脾细胞注射到受体Wistar大鼠胸腺，并以FK506短程处理受体鼠。单纯药物对照组在围手术期使用FK506但不进行胸腺内注射供体脾细胞；经典方法诱导组，腹腔注射抗淋巴细胞血清ALS(-1天) 胸腺内注射供体脾细胞(-21天)；无处理组，单纯将SD大鼠心脏移植到Wistar大鼠腹腔。观察供心存活天数、病理改变及混合淋巴细胞反应等指标的变化。结果：实验组供心存活时间较对照组明显延长，而与经典诱导组比较无统计学差异。实验组和经典诱导组的供受体脾细胞混合淋巴细胞培养各组增殖反应均较元处理组降低，而两者之间无差异。结论：心脏移植手术当日胸腺内注射供体同种抗原，同时给予短程免疫抑制剂与术前21天胸腺注射的经典诱导组同样能诱导宿主对移植物的低反应状态(免疫耐受或免疫抑制)。     

供者脾细胞对移植心脏免疫耐受的诱导

目的探讨供者脾细胞对同种异体心脏移植免疫耐受的诱导效果 ,为抗排斥反应治疗提供依据。方法分别采用供者脾细胞 (SPC)和环磷酰胺 (CP)预处理移植受者 ,然后行大鼠异位心脏移植术 ,根据实验分组对移植心脏存活情况进行观察。结果对照组、CP组和 SPC组移植心脏的存活时间分别为 7.2 1± 2 .5 6 d、9.78± 2 .5 5 d和 15 .14± 8.5 6 d,经统计学处理后证实 ,3组移植心脏的存活时间有显著性差异 (P<0 .0 5 )。结论供者脾细胞预处理移植受体 ,可以明显地延长大鼠同种异体心脏移植的存活时间。     

输注同基因骨髓间充质干细胞联合脾组织移植诱导肝移植大鼠的免疫耐受

背景:有研究显示在免疫抑制剂的作用下,同种脾脏细胞移植可诱导免疫耐受,使移植物长期存活。另有研究还显示异种骨髓间充质干细胞移植可延长移植肝存活时间。目的:观察输注与受体同基因骨髓间充质干细胞联合脾组织移植对诱导大鼠肝移植后免疫耐受的作用。方法:将受体Lewis大鼠以数字表法随机分为4组:急性排斥组行DA-Lewis大鼠原位肝移植;环孢素A组行DA-Lewis大鼠原位肝移植后灌胃给予环孢素A;干细胞组行DA-Lewis大鼠原位肝移植,同期输注异体Lewis大鼠骨髓间充质干细胞;脾组织移植组在干细胞移植组的基础上同期移植DA大鼠脾组织。观察各组生存期,肝功能情况,血清细胞因子水平,嵌合体的形成情况及肝脏病理变化。结果与结论:与其他各组相比,脾组织移植组大鼠存活时间明显延长,术后血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素、白细胞介素2、干扰素γ水平明显降低(P0.05),白细胞介素6、白细胞介素10明显升高(P0.05),30d后受体脾脏中供体阳性细胞明显升高(P0.05)。肝脏病理显示,环孢素A组和干细胞组移植肝仅呈急性轻度排斥反应,急性排斥组呈急性重度排斥反应,脾组织移植组未见明显排斥反应。说明大鼠肝脏、脾组织移植后输注同基因骨髓间充质干细胞可减轻移植肝的排斥作用,甚至诱导免疫耐受。     

口服供体脾细胞对大鼠移植肾的影响

背景：口服供体抗原诱导免疫耐受已被证实有显著效果,而脾脏为人体最大淋巴器官,富含T淋巴细胞,可提供丰富抗原。 目的：观察口服供体脾细胞对大鼠肾移植移植肾功能影响。 方法：肾移植前脾细胞组Lewis (RT11)大鼠采取口服灌胃5×105个BN (RT1n)大鼠供体脾细胞,1次/d,持续7 d;肾移植组Lewis (RT11)大鼠口服灌胃1 mL PBS为对照。 结果与结论：移植后第5天肾移植组出现移植肾排斥症状,脾细胞组平均存活时间长于肾移植组,移植后脾细胞组血肌酐、尿素氮水平升高明显慢于肾移植组;苏木精-伊红染色显示肾移植组移植肾发生急性排斥变化早于脾细胞组。说明口服供体脾细胞能诱导免疫耐受,延长移植肾存活时间。     

供体与受体移植前预处理延长大鼠移植心脏成活时间

目的：用供体品系大鼠皮肤致敏方法及腹腔内异位心脏移植模型研究雷公藤多式供体预处理及供体脾细胞受体胸腹内注射诱导免疫耐受对移植心脏存活影响及二种方法的协同效果。方法：SD大鼠作供体,Wistar大鼠作受体,所有受体在心脏移植前均无用供体皮肤致敏,A组作单纯心脏移植,B组作经供体预处理后的供心移植,C组用供体脾细胞胸腺内注射诱导耐受后作心脏移植,D组考察供受体两种预处理方法的协同效果。结果：各处理组供心存活时间均校对照组明显延长,C,D二组病检见淋巴细胞浸润及心肌坏死明显减轻,但微血管病变较严重。结论：两种方法均能明显延长移植供心成活时间,并有良好的协同效果,经胸腺途径诱导的耐受能明显抑制细胞免疫,但对急性血管排斥没有预防作用。     

供体脾细胞胸腺内注射对致敏大鼠移植心脏存活的影响

研究胸腺内耐受诱导对心脏移植存活的影响。方法ＳＤ大鼠作为供体,Ｗｉｓｔａｒ大鼠作为受体。Ａ组单做心脏移植,Ｂ组于ＨＴ前作皮肤移植致敏 ,Ｃ组在皮肤移植及ＨＴ前作供体脾细胞脾内免疫耐受诱导。结果致敏 模型能使移植排斥终点时间显著缩短,Ｃ组供心存时间显著延长并伴相应的细胞免疫排斥指标降低。     

大鼠胚胎胸腺和后肾原基联合异种移植与诱导免疫耐受的初步研究

目的 将大鼠胚胎胸腺与后肾原基联合移植至无胸腺裸小鼠,探讨联合移植能否重建受体细胞免疫功能并诱导受者对异种供者器官的特异性免疫耐受的关系.方法 从受孕第15天(E15)的Lewis大鼠胚胎中提取胸腺、后肾原基,分别植入BALB/c裸小鼠腹腔右肾包膜下及大网膜内.移植后第10周,行胸腺、后肾移植物的形态学检查及移植后肾的功能检测;并进行外周血T淋巴细胞流式细胞学检测、单向混合淋巴细胞反应(MLR)及皮肤移植试验.结果 移植后第10周,移植胸腺及后肾形态发育良好.移植后肾显示一定的排泄功能,联合移植受体双肾切除后的生存时间明显延长(P＜0.05).联合移植受体的外周血T淋巴细胞重建良好;其淋巴细胞对胸腺供体来源的Lewis大鼠脾细胞的刺激呈特异性低反应(P＜0.01);且胸腺供体来源的大鼠移植皮片平均存活时间明显大于C57BL/6小鼠、BN大鼠移植皮片存活时间(P＜0.01).结论 E15 Lewis大鼠胚胎胸腺、后肾原基联合移植至细胞免疫缺陷的裸小鼠,移植物可生长分化形成器官并发挥功能,受体裸小鼠能重建免疫功能,并有可能诱导对同源供体器官的特异性异种移植免疫耐受.     

门静脉注射供体脾细胞联合应用环孢素A诱导免疫耐受的实验研究

目的:利用大鼠异位心脏移植模型,行移植时经门静脉注射供体脾细胞联合应用环孢素A(CsA)诱导免疫耐受的实验研究,并从细胞免疫角度探讨其诱导免疫耐受机制。方法:受体鼠移植术前经门静脉注射供体脾细胞联合短期应用环孢素A(CsA)。采用MTT法检测术后受体鼠脾淋巴细胞IL-2产生水平及NK细胞活性,采用ELISA检测受体鼠IFN-γ表达水平。结果:门静脉注射供体脾细胞联合应用CsA能显著延长大鼠心脏移植物的存活期,且明显抑制受体脾淋巴细胞IL-2,IFN-γ的表达及NK细胞活性。结论:门静脉注射供体脾细胞能有效地诱导免疫耐受,干扰IL-2-NK-IFN-γ免疫网络功能可能是门静脉注射供体脾细胞诱导免疫耐受的机制之一。     

合成红藻氨酸诱发大鼠癫痫作用的研究

目的: 观察合成红藻氨酸(SKA) 诱发大鼠癫痫的作用及其作用特点。方法: Wistar大鼠40只,随机分为正常对照组、SKA 12 mg/kg、SKA 10 mg/kg 和 SKA 5 mg/kg 剂量组及红藻氨酸 (KA)10 mg/kg阳性对照组。腹腔注射给药,连续8 h观察大鼠癫痫发作的行为学变化及连续3.5 h记录其脑电图变化。结果: 合成红藻氨酸5、10、12 mg/kg 腹腔注射,可诱发大鼠癫痫发作,其行为学及脑电改变与KA对照组无明显差异。但合成红藻氨酸诱发动物癫痫呈现规律、稳定及阶段性明显的特征,且大鼠的死亡率较天然红藻氨酸低。结论: 合成红藻氨酸腹腔注射可诱发大鼠癫痫发作,以10 mg/kg为较合适剂量。     

Antitumor activity and immune enhancement of murine interleukin-23 expressed in murine colon carcinoma cells

Interleukin （IL）-23, a cytokine composed of p19 and the p40 subunit of IL-12, can enhance the proliferation of memory T cells and production of IFN-γ from activated T cells. It can also induce antitumor effects in murine model. To further evaluate the antitumor activity and immune enhancement of IL-23 in vivo, murine colon carcinoma cells retrovirally transduced with mIL-23 gene were injected subcutaneously （s.c.） into BALB/c mice. Survival time and tumor volume were observed. LDH release assay, [^3H]-TdR incorporation assay and ELISA were used to determine CTL activity, proliferation of splenocytes and level of cytokines, respectively. Number of dendritic cells （DCs） was analyzed by flow cytometry （FCM）. IL-23 secreted by Colon26/IL-23 cells suppressed the growth of tumor and prolonged the survival time of mice, enhanced proliferation of splenocytes, CTL activity, and number of DCs. IL-23 also promoted the production of Thl cytokines such as IFN-γ, IL-12 and TNF-α. However, the level of IL-4 was not enhanced significantly. These data suggested that IL-23 secreted by tumor cells can induce antitumor activitv bv enhancing immune resnonse.     

体外联合应用IL-10和甲基强的松龙处理树突状细胞诱导移植的免疫耐受

目的探讨体外联合应用白细胞介素10(IL-10)和甲基强的松龙(Medron)处理供体树突状细胞(DC)对小鼠皮肤移植术后免疫耐受的诱导效果,为抗移植术后免疫排斥反应治疗提供依据。方法以健康成年C57BL/6小鼠为供体。BALB/c小鼠为受体,随机分为11组。除A组外,其余各组均于皮肤移植前3d自尾静脉输入对应的供体DC。具体对应关系如下A组为空白对照,尾静脉输入生理盐水;B组为输入未修饰的DC;C1组为10μg/LIL-10处理组;C2组为30μg/LIL-10处理组;D1组为10mg/LMedron处理组;D2组为20mg/LMedron处理组;E1~E4组分别为10μg/LIL-10 10mg/LMedron处理组、10μg/LIL-10 20mg/LMedron处理组、30μg/LIL-10 10mg/LMedron处理组、30μg/LIL-10 20mg/LMedron处理组。同时设立F组,为BALB/c对BALB/c的同种同基因皮片移植。各组行皮肤移植术,观察受体移植皮片存活情况。结果相对于生理盐水组,E3组(30μg/LIL-10 10mg/LMedron处理组)的移植皮片存活时间最长,经统计学分析两种药物之间具有交互作用(P<0.05)。结论用IL-10和修饰的供体树突状细胞对受体进行预处理,可明显延长移植皮片的存活时间。     

Fludarabine-based conditioning for marrow transplantation from unrelated donors in severe aplastic anemia: early results of a cyclophosphamide dose deescalation study show life-threatening adverse events at predefined cyclophosphamide dose levels

Excessive adverse events were encountered in a Phase I/II study of cyclophosphamide (CY) dose deescalation in a fludarabine-based conditioning regimen for bone marrow transplantation from unrelated donors in patients with severe aplastic anemia. All patients received fixed doses of antithymocyte globulin, fludarabine, and low-dose total body irradiation. The starting CY dose was 150 mg/kg, with deescalation to 100 mg/kg, 50 mg/kg, or 0 mg/kg. CY dose level 0 mg/kg was closed due to graft failure in 3 of 3 patients. CY dose level 150 mg/kg was closed due to excessive organ toxicity (n?=?6) or viral pneumonia (n?=?1), resulting in the death of 7 of 14 patients. CY dose levels 50 and 100 mg/kg remain open. Thus, CY at doses of 150 mg/kg in combination with total body irradiation (2 Gy), fludarabine (120 mg/m(2)), and antithymocyte globulin was associated with excessive organ toxicity.     

Allogenic donor splenocytes pretreated with antisense peptide against B7 prolong cardiac allograft survival

The interaction of T cell CD28/CTLA-4 receptors with B7 on antigen-presenting cells (APCs) represents an important co-stimulatory pathway in T cell activation or anergy. Our previous study indicated that recipients immunized with allogenic donor immature dendritic cells (DCs) or resting B cells could induce specific immune tolerance and prolong allograft survival. A possible mechanism for this observation is that the expression of B7 molecules is either at a low level or lacking on these cells. The present study investigates whether blockade of B7 molecules on donor splenocytes with a B7 antisense peptide (B7AP), i.e. a peptide analogue of the CD28-binding region, could induce specific immune tolerance and prolong allograft survival in the recipients. Both the lymphocyte proliferation reaction and the mice pinna cardiac allograft experiment were performed to evaluate the role of B7AP in inducing specific immune tolerance in recipients in vitro and in vivo. The results showed that 56.65% and 20.52% of C57BL/6 splenocytes expressed B7.1 and B7.2 molecules, respectively, on their cell surface. There were no significant changes of the B7 expression on such splenocytes after being treated by the B7AP (53.28% and 19.06%, respectively). B7AP inhibited the mixed lymphocyte reaction by up to 38.4% and a dose-response correlation was observed for inhibition. The recipients (BALB/c) immunized with B7AP-pretreated C57BL/6 splenocytes induced a specific immune hypo-response (43%versus control) and notably prolonged survival of the C57BL/6 cardiac allograft by up to 20.3 days. In contrast to the normal saline group (average: 8.6 days) and FTD(10) control peptide group (<4 days), the cardiac allograft survival of the test group was extended for an additional 11.7 days. These results strongly support the notion that immunization with donor splenocytes, which had been pretreated with B7AP, induced specific immune tolerance and prolonged allograft survival in the recipients.     

联体共生诱导异基因大鼠心脏移植耐受

目的通过异基因大鼠联体共生诱导心脏移植耐受并探讨其耐受机理.方法将DA和LEW的脾细胞(2×108)分别经舌静脉注射给对方,2d后均经腹腔注射环磷酰胺(80mg/kg),第3天实施联体手术,联体1w后分开,在DA→LEW之间进行异位心脏移植手术.观察、记录移植物的存活时间,通过胸腺和脾脏内嵌合体检测、MLR、体内细胞转移、IL-2逆转实验,探讨耐受机理.结果异基因心脏移植物的存活时间明显延长;嵌合体检测显示耐受的形成和嵌合体的存在成相关性,MLR和体内细胞转移实验证明,受体大鼠的免疫应答受抑制表现为供体特异性,并且受体存在抑制细胞;IL-2逆转实验表明该耐受与克隆失活(anergy)有关.结论通过联体共生诱导大鼠耐受、延长心脏移植存活,移植耐受的形成涉及多种机制.     

Gender Differences in Pulmonary and Immune Response in Acute Experimental Endotoxicosis

Differences in the immune and inflammatory response were revealed in the lungs of male and female Wistar rats on day 1 after administration of LPS in a dose inducing the development of acute bacterial endotoxemia. Females showed more pronounced morphofunctional signs of immune system activation than males: this was characterized by more severe accidental involution of the thymus, devastation of the splenic white pulp, and enhanced production of IL-4, IL-12, and TNF-α by splenocytes. In males, production of the above cytokines decreased and inflammation in the lungs was more pronounced at these terms.     

Transplantation tolerance: the big picture. Where do we stand,where should we go?

A major goal in organ transplantation has been to safely exploit the natural processes of immune tolerance in order to minimize the dose and duration of drug immunosuppression. In this commentary, I argue that we can learn from how tumours avoid rejection, to evolve a three‐stage tolerance‐inducing strategy for transplanted tissues.     

Tamoxifen alleviates disease severity and decreases double negative T cells in autoimmune MRL-lpr/lpr mice

Previous study suggested that MRL-lpr/lpr mice treated with tamoxifen (TAM) had less severe proteinuria, reduced serum titre of anti-dsDNA autoantibodies and an increased survival rate. To investigate further the regulatory mechanisms of TAM on MRL-lpr/lpr female mice, a total dose of 200 microg per mice (5.5 mg/kg) was given every 2 weeks subcutaneously, while the control mice were injected with oil only. After being treated with TAM four times, the mice were killed and cellular functions were evaluated. The TAM-treated groups had smaller sized spleen and lymph nodes. Flow cytometric analysis of splenocytes had a significantly lower percentage of cell number of T cells and double negative T cells (CD4- CD8- T cells). There was no difference in cytokine production (interleukin (IL)-2, IL-4, IL-5, IL-10 and interferon-gamma (IFN-gamma)) from splenocytes stimulated with concanavalin A (Con A) or cytokines (IL-6) secreted by peritoneal exudate cells when stimulated with lipopolysaccharide (LPS). However, IL-2 from lymph node cells was significantly higher on TAM-treated mice. Finally, splenocytes or purified T cells stimulated with anti-CD3 antibody plus cross-linking immunoglobulin G (IgG) of the TAM-treated group had higher 3H-incorporation of proliferation assay compared with that of control groups. In vitro study further demonstrated that IL-2-activated proliferation of lymph node double negative (DN) T cells can be inhibited by TAM treatment in a dose-dependent manner. Our finding demonstrated that TAM may potentially influence T cells and modulate the immune function, which offers a novel approach to explore the feasibility of hormone therapy for autoimmune diseases.     

雷公藤甲素PG490-88联合环孢素A免疫抑制诱导肾移植急性排斥反应模型大鼠的免疫耐受

背景：近年来的研究发现雷公藤甲素具有很好的抗排异、抗肿瘤作用。PG490-88是雷公藤甲素的提取物,能够预防移植物抗宿主病和诱导免疫耐受。 目的：探讨雷公藤甲素PG490-88联合环孢素A在大鼠肾移植急性排斥反应中诱导免疫耐受的作用。 方法：改良法建立大鼠肾移植动物模型。40只Wistar大鼠做供体、40只SD大鼠做受体,随机数字表法均分为4组：对照组给予常规饮食。雷公藤甲素组给予雷公藤甲素PG490-88 20 mg/(kg•d)灌胃。环孢素A组给予环孢素A 15 mg/(kg•d)灌胃;联合治疗组：PG490-88 20 mg/(kg•d)+环孢素A 15 mg/(kg•d)灌胃。分别检测肾移植后各组1,3,5,7,14 d外周血白细胞介素2受体水平和肾移植后大鼠的脾脏淋巴细胞组织中的白细胞介素2活性蛋白表达。 结果与结论：3个治疗组的白细胞介素2活性和白细胞介素2受体均明显低于对照组(P < 0.05)。其中,联合治疗组低于雷公藤甲素组和环孢素A组,差异有显著性意义(P < 0.05)。提示,雷公藤甲素PG490-88通过对白细胞介素2、白细胞介素2受体的影响,来达到免疫抑制作用,不但具有免疫抑制作用,而且还能够诱导一定程度的免疫耐受,联合环孢素A效果会更好。