白细胞介素-10与器官移植

白细胞介素 1 0 (IL 1 0 )是近年发现的细胞因子网络中为数不多的抑制性细胞因子 ,主要由TH2细胞产生 ,能够抑制IL 2、IL 1、IFN γ、TNF α等多种细胞因子的合成及活性。越来越多的证据表明器官移植后产生免疫耐受时出现IL 1 0的高表达 ,而移植后发生排异反应时则表现为IL 1 0的低表达。移植后应用IL 1 0或将IL 1 0基因转染移植物能够延长移植物的存活。     

白细胞介素-2单克隆抗体对急性排斥反应中细胞因子表达的影响

目的　观察白细胞介素 2单克隆抗体 (IL 2Mab)对同种大鼠心脏移植物存活时间及局部细胞因子网络的影响。方法　分A组 (对照组 ) ,B组 (IL 2Mab组 )。每组 12只。分别静脉给予生理盐水及抗白细胞介素 2单克隆抗体 (A3 8 3 ) ,采用改良的移植术式建立移植模型。常规监测排斥反应发生情况。每组 5只用于观察移植物存活时间 ,7只用于动态切取标本。应用逆转录 聚合酶链反应 (RT PCR)的方法检测移植物局部细胞因子如IL 1β、IL 2、CD2 5、IL 4、IL 5、IL 6、IL 10、肿瘤坏死因子 (TNF) α、干扰素 (IFN ) γ的表达水平。结果　IL 2Mab组移植心存活时间( 2 6.4± 5 .7)d显著长于对照组 ( 8.3± 1.7)d ,实验组IL 1β、IL 5、IL 6、IL 10、IFN γ的表达较强于对照组 ,而IL 2、CD2 5、IL 4、TNF α的表达反之。结论　IL 2Mab能够显著的延长移植物生存时间 ,但对细胞因子表达模式的偏离产生的预期效果并不理想 ,其可能的机制在于细胞因子网络的复杂性。     

反复自然流产病人细胞因子水平测定

目的 :探讨不明原因反复自然流产 (RSA)病人PBMC产生IFN γ、TNF α、IL 4和血清sIL 2R水平的变化及意义。　方法 :应用酶联免疫吸附法 (ELISA)测定了 36例RSA病人、15例正常妊娠、13例经产妇以及 8例配偶白细胞免疫治疗者外周血中单个核细胞经PHA刺激后培养上清中 ,IFN γ、TNF α和IL 4以及血清中sIL 2R水平。　结果 :RSA病人PBMC产生的IFN γ为 (0 .5 9± 0 .2 6 )ng/ml,TNF α为 (1.11± 0 .5 6 )ng/ml,较对照者显著增高 (P <0 .0 5 )。经配偶白细胞免疫治疗者 (成功妊娠 )的IFN γ、TNF α水平明显低于治疗前。RSA病人妊娠后再次流产血清sIL 2R显著增高。　结论 :RSA病人某些细胞因子产生异常是妊娠的不利因素 ,sIL 2R增高与先兆流产有关。     

细胞因子基因多态性与肾移植急性排斥反应的关系

目的　探讨肾移植患者外周血中细胞因子基因多态性与急性排斥反应的关系。方法　应用聚合酶链反应测定 89例肾移植患者外周血中肿瘤坏死因子 (TNF α)、转化生长因子 (TGF β)、白细胞介素 6(IL 6)及干扰素 (IFN γ)的基因型 ,并结合HLA配型情况 ,比较各基因型别的急性排斥反应发生率。结果　在HLA DR错配的情况下 ,TNF α和IFN γ等位基因为高分泌型者 ,其术后急性排斥反应发生率较低分泌型者高 (P <0 .0 5) ;未能显示TGF β及IL 6基因表达与急性排斥反应的相关性。结论　术前检测肾移植患者细胞因子基因型 ,有助于发现可能发生急性排斥反应的高危人群 ,据此可制定合理的个体化免疫抑制治疗方案     

无细胞异种真皮基质降解因素的研究

目的　探讨影响异种无细胞真皮基质 (ADM )降解吸收的相关因素及其与炎症反应之间的关系。方法　猪和兔断层中厚皮片经Trypsin等脱细胞 ,分别制成异种 /异体ADM (xeno /allo ADM ) ,再按预处理方法不同分组 :戊二醛交联的xeno ADM (A)组 ;网状打孔和交联的xeno ADM (B)组 ;xeno ADM (C)组和allo ADM (D)组 ,分别埋植于兔皮下 ,术后 4～ 32周作大体观察和组织学检查。结果　埋植术后 4～ 12周期间 ,A组和B组可见较多的异物巨细胞 ,各组炎症反应和移植物被降解 /同化程度依次为 :C >B >A >D(P <0 .0 5～ 0 .0 0 1) ,二者之间有显著的正相关 (n =84,r =0 .185 ,P <0 .0 1)。结论　xeno ADM生物相容性远不及allo ADM ,其降解除受戊二醛交联作用和网状打孔因素影响外 ,还与局部血运和所受应力有关 ,其中炎症 免疫反应可能是降解最直接的原因。     

基底膜对异种脱细胞真皮基质移植免疫排斥反应的影响

目的比较去基底膜与保留完整基底膜的异种脱细胞真皮基质（Xeno-ADM）移植免疫学方面的差异。方法建立SD大鼠皮下脱细胞真皮基质（ADM）移植模型，120只SD大鼠随机分为4组：去基底膜组、基底膜组、自体组、空白对照组。分别将带基底膜和去基底膜的Xeno-ADM植入大鼠背部皮下，自体组分别将大鼠自体真皮基质植入大鼠背部皮下，空白对照组不植入任何材料。于术后2-32周进行组织学观察，并采用Elisa和实时定量PCR方法，检测ADM移植后血清中TNFα、IFNγ、IL-6和IgG表达水平的动态变化。结果移植早期（〈16周）基底膜组移植物炎性反应程度较去基底膜组强烈，IgG表达水平在ADM移植2周时最高，而TNFα、IFNγ、1L-6表达水平在ADM移植后8周最高，上述所有指标在移植后16周时趋向稳定，接近对照组。但3个实验组的TNFα、IFNγ、IL-6和IgG表达水平高低依次为：基底膜组〉去基底膜组〉自体组。结论移植早期（〈16周），去基底膜的Xeno-ADM移植优于带基底膜的Xeno-ADM移植。基底膜成分是Xeno-ADM移植早期引发宿主免疫反应增强的一个因素。     

大鼠烧伤后库普弗细胞在促炎细胞因子产生中的作用

目的　观察大鼠严重烧伤后早期 ,库普弗细胞在肿瘤坏死因子α(TNFα)、白细胞介素(IL) 1β、IL 6产生中的作用。方法　观察 (1)烧伤血清对体外培养的大鼠库普弗细胞分泌TNFα、IL 1β、IL 6的刺激作用 ;(2 )烧伤后大鼠库普弗细胞的细胞因子mRNA表达变化 ;(3)应用库普弗细胞特异性抑制剂三氯化钆后 ,烧伤大鼠血浆内细胞因子含量变化。　结果　烧伤血清能刺激库普弗细胞释放TNFα、IL 1β、IL 6 ;大鼠烧伤后库普弗细胞TNFα、IL 1β、IL 6mRNA表达量显著升高 ;预先抑制库普弗细胞的活性 ,烧伤后血浆TNFα、IL 1β、IL 6水平均显著降低 ,分别为烧伤组的 34.71%、36 99%、33.70 %。结论　库普弗细胞是大鼠烧伤后血浆中TNFα、IL 1β、IL 6的主要来源     

异种或同种异体脱细胞真皮基质移植后免疫细胞分型的研究

目的　比较异种脱细胞真皮基质 (Xeno ADM)与同种异体脱细胞真皮基质 (Allo ADM)移植物免疫学反应的差异。　方法　选用健康小猪和健康自愿者的中厚皮 ,分别制成Xeno ADM和Aoll ADM后 ,与深度烧伤患者的断层超薄自体皮 (UTS)重叠 ,即时覆盖其切痂创面 ,相应设为Xeno组(2 6例 )和Aoll组 (10例 )。于另 8例深度烧伤患者切痂创面上移植单纯断层自体中厚皮 (TTS) ,设为对照组。采用免疫组化方法 ,于移植术后进行组织学观察 ,并检测外周血和移植物内免疫细胞分型情况。　结果　与对照组比较 :(1)移植术后 ,Xeno组和Allo组外周血免疫细胞分型情况无明显改变(P >0 .0 5 )。 (2 )Xeno组移植物局部长期存在炎症 免疫反应 ,其中 80 %以上为CD3+ CD4 + 和CD4 5RO+ 细胞 ,CD8+ 细胞、Vs8C+ 浆细胞、CD5 7+ 的自然杀伤细胞 (NK)相对较少 ;另可见嗜酸性粒细胞和CD6 8+ CD4 + 异物巨细胞反应 ,在发生排异反应前的Xeno ADM中尤为明显 (P <0 .0 5～ 0 .0 1)。Allo组仅在移植早期 (术后 8周以内 )有中度炎症 免疫反应。　结论　人体对脱细胞真皮基质的特异性免疫反应 ,可能是在单核 巨噬细胞参与下、以CD4 + T淋巴细胞介导的细胞免疫为主要表现 ;辅助性T细胞和巨噬细胞构成的异物巨细胞可能在其中起重要作用     

自体皮与异种或异体脱细胞真皮基质复合移植的实验观察

充分有效的真皮替代物(DS)是复合皮(CS)在临床推广应用的基础。为拓宽DS的来源和应用范围,笔者单位在以往的研究基础上,将超薄自体皮片(UTS)、自体表皮和微粒皮分别与戊二醛交联的异种或异体脱细胞真皮基质(xeno /allo ADM)重组形成CS,观察各组皮片成活情况,并探讨两种ADM的炎症免疫反应和组织降解的差异。一、材料与方法1.ADM的制备:allo ADM来自2月龄、体重10～15kg的健康小白猪,xeno ADM来自术后剩余的正常皮肤,参照文献[1]制备。组织学观察证实真皮内无细胞残留。2 .CS移植方法:在猪脊背两侧分别设计6 .0cm×6.0cm创面4块。…     

正常妊娠与自然流产小鼠模型母胎界面细胞因子表达特征的比较

目的　比较正常妊娠与自然流产模型母胎界面细胞因子表达特征的差异 ,探讨T辅助细胞 (Th) 1型和 2型 (Th1 /Th2 )细胞因子在介导流产发病中的作用机制。　方法　建立正常妊娠模型CBA×BALB/c和自然流产模型CBA×DBA/ 2。采用酶联免疫吸附试验 (ELISA)测定两组模型孕第 9和 1 4天母胎界面组织培养上清中Th1 /Th2型及相关细胞因子表达水平 ;以白细胞介素 4/干扰素 γ(IL 4/IFN γ)比值作为衡量Th2 /Th1型免疫调节平衡的指标。　结果　孕第 9天 ,与正常妊娠模型相比 ,自然流产模型母胎界面IL 4、IL 1 0和转化生长因子 (TGF) β2的表达显著降低 (P <0 .0 5) ;IFN γ、肿瘤坏死因子 (TNF) α的表达显著升高 (P <0 .0 5) ,IL 4/IFN γ比值显著降低 (P <0 .0 5)。至孕第 1 4天 ,自然流产模型母胎界面IFN γ、TNF α表达仍显著高于正常妊娠模型 (P <0 .0 5) ,IL 1 0表达、IL 4/IFN γ比值仍低于正常妊娠模型 (P <0 .0 5) ,IL 4和TGF β2在两组模型中的表达无显著差异。两组模型孕第 9和 1 4天母胎界面均未测到IL 2的表达。　结论　与流产模型相比 ,无论在妊娠早期或中晚期 ,正常妊娠模型母胎界面局部均呈现明显的Th2型免疫偏倚。母胎界面Th1 /Th2型细胞因子表达调节失衡是导致流产的重要原因之一     

复合皮移植的实验研究与临床应用

目的寻求一种较理想的真皮替代物,用以修复全厚皮肤缺损创面。方法(1)将异体／异种脱细胞真皮基质(allo／xeno-ADM) 自体刃厚皮组成复合皮(CS),以自体刃厚皮作对照,采用一步移植法进行实验研究和临床研究。移植术后应用生物化学及免疫学方法观察移植物抗原性的改变、表皮一真皮连接区(DEJ)基底膜带结构的再形成情况。(2)在此基础上,应用前述CS移植技术治疗53例全厚皮缺损或瘢痕切除患者,观察术后皮片成活情况并作随访。结果CS一步法移植后的成活情况良好。(1)移植后28周可见DEJ基底膜带重新形成,基底膜细胞呈极性生长并呈波浪形排列,真皮乳头与皮钉再生。allo-ADM的抗原性明显低于xeno-ADM,仅在术后早期有少数炎性细胞浸润,未见明显的免疫排斥反应。(2)移植于53例患者的70块CS中,65块完全存活占92．9％,2块因更换敷料不当部分存活,3块因创面感染移植失败。患者最长随访时间为8．5年,所移植的CS质地和色泽接近正常皮肤。结论allo／xeno-ADM的抗原性很低,移植后能发挥长期的真皮支架或诱导组织再生的真皮“模板”作用。     

异种脱细胞真皮基质抗原性的实验研究

目的　观察不同方法制备的异种脱细胞真皮基质 (Xeno -ADM )埋植物的抗原性差异。　方法　将猪断层中厚皮片采用胰蛋白酶 +tritonX 10 0等处理 ,制成Xeno -ADM ,分为戊二醛交联的Xeno ADM1组、网状交联的Xeno ADM2 组、未交联的Xeno ADM3 组、动物先用Xeno ADM蛋白致敏后再移植交联的Xeno ADM4组和未交联的异体兔Allo ADM对照组。分别将移植物埋植于 5组共计 2 5只日本白兔的耳部和背部皮下 ,术后 2～ 32周检测兔血清抗ADMs抗体效价 ,并观察移植物大体和组织学变化。　结果　Xeno ADM4埋植物组血清抗Xeno ADM蛋白抗体效价最高 ;排除致敏因素的影响 ,Xeno ADM各组血清抗体滴度依次为 :Xeno ADM3 组 >Xeno ADM2 组 >Xeno ADM1组 (P<0 .0 5～ 0 .0 0 1) ;在Allo ADM组中 ,有 4 0 %的血清样本呈现出抗Allo ADM蛋白抗体阳性。组织学上 ,Xeno ADM移植部位有激烈而持久的炎症反应 ,明显强于Allo ADM异体埋植对照组 ;ADM被降解吸收的程度依次为 :Xeno ADM3 >Xeno ADM2 >Xeno ADM4>Xeno ADM1>Allo ADM (P <0 .0 5～ 0 .0 0 1) ,在交联型ADM组易产生异物巨细胞反应。　结论　Xeno ADM的免疫原性强于Allo ADM ,诱导宿主产生IgG限制的免疫反应 ;大张变性的ADM埋植物能降低抗原 抗体反应 ,减缓炎症免疫反应导致的ADM     

Role of blood glucose in cytokine gene expression in early syngeneic islet transplantation

In islet transplantation, local production of cytokines at the grafted site may contribute to the initial nonspecific inflammation response. We have determined whether the metabolic condition of the recipient modulates the cytokine expression in islet grafts in the initial days after transplantation. Normoglycemic and hyperglycemic streptozotocin-diabetic Lewis rats were transplanted with 500 syngeneic islets, an insufficient beta cell mass to restore normoglycemia in hyperglycemic recipients. The expression of IL-1beta, TNF-alpha, IFN-gamma, IL-6, IL-10, and IL-4 genes was determined by real-time PCR in freshly isolated islets, in 24-h cultured islets and in islet grafts on days 1, 3, and 7 after transplantation. IL-1beta mRNA was strongly and similarly increased in normoglycemic and hyperglycemic groups on days 1, 3, and 7 after transplantation compared with freshly isolated and cultured islets. TNF-alpha mRNA was also strongly increased on day 1, and it remained increased on days 3 and 7. IL-6 and IL-10 were not detected in freshly isolated islets, but their expression was clearly enhanced in 24-h cultured islets and islet grafts. IL-6 was further increased in hyperglycemic grafts. IL-10 expression was increased in both normoglycemic and hyperglycemic grafts on day 1 after transplantation, and remained increased in hyperglycemic grafts compared to 24-h cultured islets. IFN-gamma mRNA was barely detected in a few grafts, and IL-4 mRNA was never detected. Thus, the inflammatory response in islet grafts was maximal on day 1 after transplantation, it was sustained, although at lower levels, on days 3 and 7, and it was partly enhanced by hyperglycemia.     

Intestinal graft versus native liver cytokine expression in a rat model of intestinal transplantation with and without donor-specific cell augmentation

BACKGROUND: Immunomodulatory strategies such as donor-specific bone marrow or blood transfusions have been used to promote engraftment after intestinal transplants. We previously showed that delivery of donor antigen via the portal vein can effectively reduce the rate of intestinal graft rejection. The purpose of our current study was to investigate the impact of donor-specific cell augmentation (blood versus bone marrow) via the portal vein on cytokine expression in intestinal grafts versus native livers. MATERIAL AND METHODS: We performed heterotopic small intestinal transplants between male Brown-Norway (donor) and female Lewis (recipient) rats. We studied 10 groups according to the type of donor-specific cell augmentation and the use and dose of immunosuppressive therapy. For cell augmentation, donor-specific blood or bone marrow was transfused via the donor portal vein immediately before graft implantation. For immunosuppression, tacrolimus was used post-transplant at a high or low dose. Control rats received neither immunosuppression nor cell augmentation. Tissue samples for histological assessment were obtained at designated time points. RNA was extracted from intestinal graft and native liver biopsies for cytokine measurements (IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-5, IL-6, IFN-gamma, TNF-alpha, and TNF-beta). Chimerism levels were determined using Q-PCR analysis. RESULTS: Without concurrent immunosuppression, neither portal donor-specific blood nor bone marrow transfusion reduced the rate of rejection. With immunosuppression, outcome was significantly better after portal donor-specific blood (versus bone marrow) transfusion. Irrespective of the type of donor-specific cell augmentation, severe rejection caused strong cytokine expression in the grafts of IL-1 alpha, IL-1 beta, IFN-gamma, and TNF-alpha; in the native livers, mainly of TNF-alpha (with IFN-gamma showing hardly any increase). In general, rejection caused stronger cytokine expression in the grafts than in the native livers. Mild rejection correlated well with strong intragraft expression of IL-6, TNF-alpha, and TNF-beta (early rejection markers); severe rejection with IL-1 alpha, IL-1 beta, IFN-gamma, and TNF-alpha (late rejection markers).In addition to cell augmentation per se, the type of cell augmentation also had an impact on cytokine expression in both grafts and native livers. Cell-augmented (versus tacrolimus-treated) rats showed hardly any differences in intragraft cytokine expression, but the expression of almost all cytokines was significantly stronger in the native livers. With immunosuppression, bone marrow infusion increased intragraft cytokine expression of IL-1 alpha, IL-1 beta, IFN-gamma, and TNF alpha, as well as liver cytokine expression of IL-1 beta, compared to blood transfusion. This finding reflected the more advanced rejection stages in the bone marrow infused group; different types of donor-specific cell augmentation had similar effects on liver cytokine expression. In the absence of myoablative therapy, chimerism levels were low, in both cell-augmented and non-cell-augmented groups. CONCLUSIONS: Rejection and donor-specific cell augmentation independently causes differences in intragraft versus native liver cytokine expression after intestinal transplants. Portal donor-specific blood transfusion, as compared with donor-specific bone marrow infusion, lowered the incidence of rejection and diminished intragraft cytokine up-regulation.     

含银异种脱细胞真皮基质的实验研究

目的了解含银异种脱细胞真皮基质(Xeno-ADM)的多项生物学性状,观察其移植效果。方法制备单纯xeno-ADM,再用2 g／L硝酸银浸泡,制成含银xeno-ADM。检测两种xeno-ADM对笔者单位烧伤患者创面常见菌的抑菌效果,并进行组织学观察;测量含银xeno-ADM的Ag+含量。在27只家兔背部制作全层皮肤缺损创面,分为A、B、C组,每组9只。A组移植自体刃厚皮,B组移植单纯xeno-ADM+自体刃厚皮,C组移植含银xeno-ADM+自体刃厚皮。术后2、4、6周取移植部位皮肤标本作形态学观察,并计算创面收缩率;术后2周计算移植皮片(未)成活率,并检测各组家兔淋巴细胞增殖活性。结果(1)含银xeno-ADM对创面常见菌的抑菌效果明显优于单纯xeno-ADM (P<0．05)。两种xeno-ADM中的表皮已完全除去,胶原纤维粗细均匀、排列规则、无明显变性,真皮中无细胞及细胞碎片。含银xeno-ADM的Ag+含量为(2．7±0．7)mg／g。(2)术后6周,A组家兔移植皮片呈暗红色,挛缩明显,易破溃,胶原纤维排列紊乱;B、C组移植皮片颜色接近周围正常皮肤,光滑无瘢痕,质地良好,胶原纤维排列有序,表皮-真皮连接结构和基底细胞桥粒、半桥粒结构以及基底膜重建明显。术后2、4、6周,A组家兔创面收缩率均明显高于B、C组(P<0．05);B、C两组创面收缩率相似(P>0．05)。术后2周,C组皮片完全成活率为91．7％,显著高于A组(77．8％)及B组(80．6％);3组家兔淋巴细胞增殖活性相似(P>0．05)。结论含银xeno-ADM脱除了基质中有免疫原性的细胞成分,保留了组织的基本结构和完整的胶原纤维支架,且具有较好的局部抗菌效果,不失为一种良好的真皮替代物。     

Comparison of cytokine levels in bilateral ear effusions in patients with otitis media secretoria.

OBJECTIVE: The aim of this study was to clarify the site of primary pathology in otitis media with effusion. STUDY DESIGN AND SETTING: The levels of the inflammatory mediators TNF-alpha, TNF-beta, IL-1beta, IL-8, IL-6, IL-4, IL-2, IL-5, IL-10, and IFN-gamma were measured in 54 pairs (108 samples) using specific enzyme-linked immunosorbent assays (ELISAs). RESULTS: The levels of pro-inflammatory cytokines TNF-alpha, TNF-beta, IL-1beta, IL-8, anti-inflammatory cytokines IL-5, IL-4, IL-10, IL-6, and cytokines with immunoregulatory potential IFN-gamma, IL-2 were different between both ears of the same patient. CONCLUSION: The results suggest that in one individual both ears have different immunological processes or rates and this has implications on using the opposite ear as a control in clinical trials. SIGNIFICANCE: Profiles of interlink of examined cytokines within the samples of both ear effusions were significantly different. A significant bilateral difference was found in the levels of IFN-gamma.     

TNF-alpha, IL-6, IFN-gamma, and IL-10 gene expression polymorphisms and the IL-4 receptor alpha-chain variant Q576R: effects on renal allograft outcome.

BACKGROUND: There has been much interest recently in the effects of various cytokine gene expression polymorphisms on graft outcome. However, the results of these investigations reveal the outcomes to be organ-specific and center-specific. We sought to confirm and add to some of the earlier findings by studying the impact of tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), interferon-gamma (IFN-gamma), and interleukin-6 (IL-6) polymorphisms and the interleukin-4 (IL-4) receptor alpha-chain variant on posttransplant renal allograft outcome. METHOD: TNF-alpha, IL-6, IFN-gamma, and IL-10 gene promoter region polymorphisms were assayed genotypically by PCR-SSP on 120 patients transplanted at the Albany Medical Center. These patients were also typed for the IL-4 receptor alpha-chain variant Q576R. RESULTS: Producers of high levels of the proinflammatory cytokine TNF-alpha were found to be at increased risk for acute rejection episodes if the allograft was mismatched for the molecular products of the class II region of the human major histocompatibility complex (HLA-DR). Expression level polymorphisms of the IL-6, IFN-gamma, and IL-10 genes were not associated with acute rejection episodes, nor was the IL-4 receptor alpha-chain variant Q576R. CONCLUSIONS: These data would suggest that the production of high levels of the cytokine TNF-alpha is especially detrimental to graft survival when the recipient's T-helper lymphocytes are being activated by mismatched donor HLA-class II antigens. Typing all potential kidney recipients for TNF-alpha, and providing well-matched organs for high producers of this cytokines, may be expected to increase rejection-free graft survival in these patients.