同种脾移植治疗血友病甲五例报告

为５例重症血友病甲患者施行了同种全脾移植，３例为尸体供脾，２例为母亲供脾。１例因脾蒂扭转于手术当天丧失移植脾，余４例术后第Ⅷ因子（ＡＨＧ）显著升高。１例术后３４天因严重移植物抗宿主反应（ＧＶＨＲ）而切脾，另３例移植物有功能超过１年，其中１例至今已超过３年，生活正常。本组病例在数量和疗效上均居国际领先水平。结果提示，脾是产生ＡＨＧ的重要器官之一，脾移植（特别是亲属脾移植）有可能使血友病甲获根治，尸脾移植的排斥和ＧＶＨＲ强烈，是导致失败的主要原因，供受体术前预处理可预防和减轻排斥及ＧＶＨＲ。     

临床小肠移植并发症及其防治

小肠移植由于排斥，ＧＶＨＤ，感染等并发症，临床上直到１９８８年才有首例成功。本文着重综述临床小肠移植中排斥、ＧＶＨＤ、感染以及技术等并发症的原因和诊治。     

造血干细胞移植的进展和展望

免疫抑制剂在造血干细胞移植 (ＨＳＣＴ)中用于移植物抗宿主病 (ＧＶＨＤ)的预防及治疗 ,近年来不断有新药推出。霉酚酸酯 (ＭＭＦ)作为一种新的免疫抑制剂 ,与环孢素Ａ (ＣｓＡ)和氨甲喋呤 (ＭＴＸ)联合用于ＧＶＨＤ的预防 ,国外已有报道 ,国内广州省人民医院、浙江大学医学院附属第一医院、北京大学血液病研究所几乎同时开始应用 ,并取得了较好的效果。他克莫司 (ＦＫ5 0 6 )体外研究显示其免疫抑制作用较ＣｓＡ强。临床显示ＦＫ5 0 6 ＭＴＸ对急性ＧＶＨＤ预防有效。但其效果尚需更多病例证实。为减少ＧＶＨＤ的发生 ,除了免疫抑制剂 ,…     

亲属脾移植术后移植物抗宿主反应和移植脾亢的成功治疗经验

移植物抗宿主反应（ＧＶＨＲ）和移植脾亢是脾移植的两个棘手问题，因二者来势凶猛，控制困难，既往治疗切除移植脾脏。我们介绍一例你亲供脾移植术后天和２１天发生ＧＶＨＲ和移植脾亢，分别采用深部Ｘ线及钴６０局获得成功的治疗经验。     

他克莫司在重度急性移植物抗宿主病治疗中的应用

异基因造血干细胞移植是治疗恶性血液病及再生障碍性贫血等疾病的有效方法[1] ,但这种移植受到供、受者ＨＬＡ配型的限制 ,即使在ＨＬＡ完全相合的供、受者之间移植 ,仍然有 5 0 %左右的受者术后发生急性移植物抗宿主病(ＡＧＶＨＤ) ,而其中又有约一半的受者最终死于本病或本病的免疫抑制治疗。因此 ,ＡＧＶＨＤ是异基因造血干细胞移植的主要并发症 ,它的预防及治疗效果是移植成功与否的关键。近 1年来 ,我们将免疫抑制剂他克莫司 (ＦＫ5 0 6 )用于 5例难治性重度 (Ⅳ度 )ＡＧＶＨＤ的治疗中 ,取得较满意的结果。报告如下。一、资料与方法1.…     

HLA空间构象的模拟比较

通过对两对骨髓移植后发生移植物抗宿主病（ＧＶＨＤ）的供受体有差异的ＨＬＡⅠ类抗原的结构模拟和比较，发现差异ＨＬＡ抗原在空间构象上存在明显差异，并表现在分子表面上，这种蛋白质分子表面空间构象上的差异是移植后ＧＶＨＤ的引发因素。研究结果提示移植供受体差异ＨＬＡ蛋白质的模拟比较可以用于预测ＧＶＨＤ的发生。     

中华眼镜蛇蛇毒对大鼠异种心脏移植超急性排斥反应的影响

试用中华眼镜蛇蛇毒消耗补体，观察其对豚鼠到ＳＤ大鼠异种心脏移植超急性排斥反应的影响。一次性给予小剂量ＣＣＶ腹腔注射后可明显降低大鼠血清补体溶血活性，４－２４小时后渐降至给药前１／２以下水平。ＣＣＶ可明显延长异种移植心脏４ 存活时间达２．５－３６小时，对照组仅为２５分钟；ＣＣＶ与脾切除，前列腺素Ｅ１联用可进一步延长移植心的存活时间，最长１例达４０小时。     

应用环孢素A治疗异种肝细胞移植的排斥反应

目的　探讨应用环孢素A(CsA)治疗异种肝细胞移植排斥反应的疗效及其排斥机制。方法　将豚鼠肝细胞植入D-氨基半乳糖诱导的肝衰大鼠脾内,分CsA治疗组和单纯移植组。观察2周生存率及脾病理组织学检查;酶联免疫双抗体夹心法(Sandwich-ELISA)测定大鼠血清白细胞介素(IL)-2的浓度。结果　2周生存率比较CsA治疗组（78.6%）与单纯移植组（80.0%）相比差异无显著性(P＞0.05)。应用CsA可减缓炎症细胞浸润。血清IL-2浓度检测正常大鼠为(28.7±7.0)ng/L,移植后各组血清IL-2浓度变化不大,直至移植后第7天,CsA治疗组为(28.5±6.0)ng/L、单纯移植组为(31.5±8.0)ng/L。3组间差异均无显著性(P＞0.05)。结论　在异种肝细胞脾内移植中,细胞免疫发生的较迟和/或较弱。单独应用CsA治疗异种肝细胞脾内移植引发的排斥反应其疗效不佳。     

肝细胞移植治疗急性肝功能衰竭的免疫排斥研究

目的研究肝细胞腹腔移植治疗急性肝功能衰竭的免疫排斥反应。方法用胶原酶灌注法分离幼猪及BALB/c和C57BL/6小鼠肝细胞;用腹腔注射CCl4的方法建立C57BL/6急性肝功能衰竭模型;将实验动物分为同基因移植组、同种异基因移植组和异种移植组。各组动物肝细胞移植入急性肝功能衰竭小鼠腹腔内,观察受体小鼠存活情况,比较各组动物T细胞亚群、免疫球蛋白水平和细胞因子的变化。结果①受体小鼠存活情况:同基因移植组为8/10,同种异基因移植组为6/10,异种移植组为3/10,同基因移植组和同种异基因移植组受体的生存情况明显好于异种移植组(P<0.05)。②T细胞亚群变化:移植后7 d内,同基因移植组外周血中CD4+和CD8+T细胞均无明显变化,同种异基因移植组CD4+T细胞于移植后3 d时达高峰(P<0.05),而CD8+T细胞7 d内无明显变化,异种移植组CD4+和CD8+T细胞移植后7 d内均明显升高,且于移植后3 d时达高峰(P<0.05)。③免疫球蛋白水平:同基因移植组血清免疫球蛋白IgM和IgG水平在移植后0.5、1和3 d时未见明显变化,同种异基因移植组和异种移植组的IgM在移植后1 d时达高峰(P<0.05),而IgG在移植后3 d时达高峰(P<0.05)。④血清细胞因子水平:移植后7 d,同种异基因移植组和异种移植组的IFN-γ、TNF-α及IL-2血清浓度明显高于同基因移植组(P<0.05);异种移植组的IL-6血清浓度明显高于同基因移植组和同种异基因移植组(P<0.05)。结论无论同种还是异种肝细胞移植,初期均发生了强烈的CD4+及CD8+T细胞参与的细胞免疫和较强体液免疫反应。     

同种大鼠带血管异位脾移植模型的建立

目的　建立同种大鼠带血管异位脾移植的实验模型。方法　远交系Wistar大鼠间行同种大鼠带血管异位脾移植。供脾带门静脉段和腹主动脉段分别与受体的下腔静脉和腹主动脉端—侧吻合 ,供脾异位移植于升结肠旁沟 ,与侧腹壁行三点固定。结果　手术成功率为 69 2 % (18/ 2 6例 ) ,术后 1周存活率为 61 1% (11/ 18例 ) ,2例大鼠获长期存活 (>90d)。脾蒂无一例扭转。组织学检查证实术后早期移植脾存活 ,发生急性排斥反应的时间平均为 3d。结论　建立同种大鼠带血管异位脾移植模型是可行的     

移植物抗宿主病与异种移植研究综述

目的对移植物抗宿主病的发生条件、发病机制、疾病特点及检测方法的研究进行简要综述。方法对近年来国内外有关文献进行检索,总结。结果移植物抗宿主病的发生需要一定的条件,细胞及体液免疫均参与该病的发病过程,根据其特点可用相应的监测方法进行诊断和分级。结论作为移植物与宿主相互作用的一种形式,移植物抗宿主病对宿主可产生严重危害;它在异种移植中也可出现,且因能用于异种移植免疫、肿瘤免疫等研究,正逐渐受到重视。     

Comparison of activation requirements and activation phenotype of allogeneic and xenogeneic rodent responses in vivo and in vitro

BACKGROUND: Both discordant and concordant xenogeneic responses are dominated by humoral immunity. Recent advances in molecular engineering approaches may largely prevent rejection by means of this pathway, leaving the cellular arm of the immune response as the principal remaining barrier to successful engraftment. METHODS: To characterize further the cellular response to xenogeneic tissues, we used the intracellular fluorescent marker CFSE (5-(and-6)-carboxyfluorescein diacetate succinimidyl ester) to track the mitotic record of T cells (and T cell subsets) after either xenogeneic or allogeneic activation in vitro or in vivo. Activation marker expression was monitored by simultaneous labeling with antibodies for either CD25 or CD134. RESULTS: The in vitro and in vivo responses of Lewis lymphocytes were generally similar in magnitude and timing comparing activation with allogeneic or xenogeneic stimulators. However, the xenogeneic T cell precursor frequency was found to be markedly higher than that previously reported and were comparable to that seen in allogeneic responses. Xenogeneic responses were unique in the continued expression of activation markers in later division cycles. In addition, CD4 and CD8 T-cell proliferation was highly dependent on stimulator class II expression, highlighting the importance of CD4 T cells and the indirect pathway in the xenogeneic response. CONCLUSIONS: An unexpectedly high precursor frequency was detected for xenogeneic cellular responses in the rat anti-mouse combination and was comparable to that seen in allogeneic responses. Differences in xenogeneic versus allogeneic activation profiles exist that may result from the cellular pathways used for activation.     

Regulation of B- and T-cell mediated xenogeneic transplant rejection by interleukin 12

BACKGROUND: Xenotransplantation may provide a solution to the increasing shortage of donor organs. Acute vascular rejection and cell-mediated rejection remain the primary barriers to successful xenotransplantation. In animal models where acute vascular rejection can be attenuated, xenografts succumb to cell-mediated rejection. The mechanisms of acute vascular rejection and cell-mediated rejection are poorly understood. METHODS: Using a heterotopic rat-to-mouse cardiac transplantation model, we demonstrate that IL-12p40 attenuates both allogeneic and xenogeneic acute vascular rejection pathology by suppressing B-cell activation and anti-rat isotype switching. To study the mechanism of xenogeneic cell-mediated rejection, we use B-cell deficient mice that only develop cell-mediated rejection pathology. To elucidate the role of IL-12 in cell-mediated rejection, we generated B cell/ IL-12p40 double knockout mice. RESULTS: We demonstrate that xenogeneic cell-mediated rejection is mediated by CD4+ T cells, and is accompanied by elevated FasL and granzyme mRNA expression. Strikingly, by generating B cell/IL-12p40 double knockout mice, we demonstrate that xenogeneic cell-mediated rejection is IL-12p40 dependent. In contrast, we demonstrate that allogeneic cellular rejection is IL-12p40 independent. CONCLUSIONS: We conclude that IL-12 plays a dual role in xenotransplantation by driving xenogeneic CD4+ T cell responses but suppressing both allogeneic and xenogeneic B cell responses. Therefore, the mechanism of allogeneic and xenogeneic transplantation rejection is differentially regulated by IL-12.     

Xenogeneic ex vivo hemoperfusion of rhesus monkey livers with human blood

Abstract In order to copy the clinical situation of concordant xenotransplantation, Rhesus Monkey livers were hemoperfused with human blood. Changes of immunological (TNFα, IL-1β, IL-2, IL-2R, IL-6, IFNy, TXB2, 6kPGF1α, sICAM-1, SELAM-1, sHLA-I-Ag) and pathophysiological (GOT, GPT, LDH, CK) parameters were followed. Our experiment proves that all phenomena start in the first hour of xenogeneic blood circulation. Xenogeneic rejection in our concordant system is surprisingly severe. Preformed natural antibodies only cannot be the reason of such a damage [5, 6]. We think that beside other important immunological mechanisms, humoral mediators play a considerable role at the beginning of a xenogeneic rejection.     

Transplantation of mouse embryonic stem cells induces hematopoietic and tissue chimerism in rats

Embryonic stem cells (ESC) can differentiate into all cell lineages, and ESC-like cells were shown to induce hematopoietic chimerism and tolerance in allogeneic models. The aim of our study was to test the capacity of mouse ESC (mESC) to engraft in rats in a xenotransplantation setting. Forty-six rats were transplanted intravenously with 1 million mESC, without immunosuppression (group 1, n = 23) or with cyclosporine (group 2, n = 23). Three months after mESC transplantation, skin grafts were performed from allogeneic, xenogeneic identical to mESC, or xenogeneic third party donors. At day 27 post-transplant, we detected circulating mouse cells in the blood of 4/23 and 5/23 animals of group 1 and group 2, respectively. Chimerism was confirmed by PCR. We also identified long-term surviving murine cells within livers of chimeric animals. Skin grafts showed no difference in survival between allogeneic and xenogeneic donors. Transplantation of xenogeneic mouse ESC induced short-term chimerism in the blood and persistent tissue chimerism in the liver of recipient rats, but did not induce tolerance to skin grafts. Improved immunosuppressive protocols should be tested to prolong chimerism and allow tolerance.     

异种肝细胞移植缓解大鼠急性肝功能衰竭及其排斥反应观察

目的探讨异种肝细胞移植对大鼠急性肝功能衰竭防治效果及其免疫排斥反应。方法将异种豚鼠肝细胞,90%肝除术前1d植入大鼠脾脏内。观察受试大鼠存活时间,及切肝术后24h血生化改变。另外观察植入肝细胞被排斥情况及受体CH50、异种抗体IgG、IgM水平变化。结果(1)中位存活时间,对照组为21h,同种组为56h,异种组为40h。同种组存活时间较对照组延长(P<0·01),异种组亦延长(P<0·05)。(2)异种组血糖和凝血酶原时间改善较明显(P<0·05),同种组谷丙转氨酶、总胆红素、血糖、凝血酶原时间都有明显改善(P<0·05或P<0·01)。(3)受体CH50和异种抗体IgM水平下降与植入异种肝细胞排斥过程同步。结论异种肝细胞移植对大鼠急性肝功能衰竭有防治作用,补体和异种抗体IgM与排斥反应关系密切。     

T cells from presensitized donors fail to cause graft-versus-host disease in a pig-to-mouse xenotransplantation model

BACKGROUND: The ability of T cells from pigs, the most suitable donors for clinical xenotransplantation, to induce graft-versus-host disease (GVHD) and to facilitate hematopoietic cell engraftment in highly disparate xenogeneic recipients remains unclear. In this article, the authors address these questions in a presensitized pig-to-mouse transplantation model using porcine cytokine-transgenic mice. METHODS: Swine donors were presensitized by mouse skin grafting and boosted by the injection of mouse cells after the skin graft was rejected. Porcine peripheral blood mononuclear cells (PBMC) and splenocytes were collected at various times after mouse skin grafting, and their potential to induce GVHD and to facilitate donor hematopoietic cell engraftment in conditioned murine recipients was evaluated. RESULTS: Presensitization of donor pigs resulted in marked enhancement of anti-mouse responses, as reflected in augmented anti-mouse mixed lymphocyte responses, cell-mediated cytotoxicity, and antibody production. However, injection of high numbers of PBMC and splenocytes (>1 x 10(8)) with bone marrow cells from the presensitized pigs failed to induce detectable GVHD or long-term chimerism in mice that were treated with depleting anti-T-cell and natural killer cell antibodies, cobra venom factor, medronate-liposomes, and 4 Gy of whole-body and 7 Gy of thymic irradiation. Histologic analysis revealed no mononuclear cell infiltration or GVHD-associated lesions in the liver, intestine, lungs, or skin of the mouse recipients. Furthermore, the recipient mice had no detectable T cells or anti-pig immunoglobulin G antibodies in the blood by 6 weeks after injection of porcine cells. CONCLUSION: These results demonstrate that porcine T-cell function is severely impaired in the xenogeneic murine microenvironment.     

Hemolymph transfer as an assay for immunorecognition in the cockroach Periplaneta americana and the locust Schistocerca gregaria

A quantitative assay for measuring the number of hemocytic nodules formed in response to foreign particles and soluble molecules has been used, in the locust Schistocerca and the cockroach Periplaneta, to investigate the response to transferred hemolymph. Xenogeneic test particles, rabbit neutrophil leukocytes, stimulate formation of nodules when injected into both insect species, compared with saline-injected controls. However, the number of nodules formed in the locust in response to cockroach hemolymph is significantly reduced compared with the response to other xenogeneic cells, and it is suggested that, in view of the strong reactivity of cockroach hemocytes to locust hemolymph and plasma, a graft-versus-host response might be occurring in the recipient locust. Whole hemolymph transferred allogeneically between Periplaneta, or xenogeneically from Blatta to Periplaneta, does not stimulate a response in the recipient. This corresponds well with results from other assays for immunorecognition of transplants and is further confirmation that allogeneic and, in some combinations xenogeneic, recognition is absent in insects.     

Comparison of in vivo lymphocyte proliferation between allogeneic and xenogeneic heart transplantation in mice

There are controversial in vitro data comparing the strength of the cellular immune response between allogeneic and xenogeneic stimulator/responder combinations. The present study therefore compares in vivo lymphocyte proliferation using heart transplantation (HTx) models in mice. Heterotopic HTx into BALB/c mice was performed using donor organs from mice (BALB/c and C57BL/6) or Lewis rats. Intraperitoneally given bromodeoxyuridine (BrdU) was incorporated into the DNA and was subsequently analyzed by flow cytometry. On postoperative days 3 and 5, proliferation of splenocytes, CD4(+) T-lymphocytes, and CD19(+) B-lymphocytes was significantly higher after xenogeneic than after allogeneic and isogeneic HTx. No significant difference was observed when proliferation of CD8(+) lymphocytes was determined. The increased in vivo proliferation after xenotransplantation may reflect an earlier and probably stronger cellular immune response compared to allogeneic transplantation. The higher CD4(+) lymphocyte proliferation underscores the importance of this cell population in xenograft rejection.     

Anti-mouse CD154 antibody treatment facilitates generation of mixed xenogeneic rat hematopoietic chimerism, prevents wasting disease and prolongs xenograft survival in mice

BACKGROUND: The induction of xenogeneic hematopoietic chimerism is an attractive approach for overcoming the host response to xenografts, but establishing xenogeneic chimerism requires severe myeloablative conditioning of the recipient. The goal of this study was to determine if co-stimulation blockade would facilitate chimerism and xenograft tolerance in irradiation-conditioned concordant recipients. METHODS: Wistar Furth rat bone marrow (BM) cells were injected into irradiation-conditioned C57BL/6 mice with or without co-administration of anti-mouse CD154 monoclonal antibody (mAb). Chimerism was quantified by flow cytometry, and mice were transplanted with WF rat skin and islet xenografts. RESULTS: Blockade of CD40-CD154 interaction facilitated establishment of xenogeneic chimerism in mice conditioned with 600 cGy irradiation. Anti-CD154 mAb was not required for establishment of chimerism in mice treated with 700 cGy. However, mice irradiated with 700 cGy but not treated with anti-CD154 mAb developed a "graft-versus-host disease (GVHD)-like" wasting syndrome and died, irrespective of their development of chimerism. Xenogeneic chimeras established with irradiation and anti-CD154 mAb treatment exhibited prolonged skin and, in many cases, permanent islet xenograft survival. Chimerism was unstable and eventually lost in most recipients. Skin xenografts were rejected even in mice that remained chimeric, whereas most islet xenografts survived to the end of the observation period. CONCLUSIONS: Blockade of host CD40-CD154 interaction facilitates the establishment of xenogeneic chimerism and prevents wasting disease and death. Chimerism permits prolonged xenograft survival, but chimerism generated in this way is unstable over time. Skin xenografts are eventually rejected, whereas most islet xenografts survive long term and perhaps permanently.