Abstract: Cardiac transplantation is an effective therapeutic method for terminalstage heart diseases. The immunosuppressive treatment based on calcineurin inhibitors (CsA and FK506) is most commonly used, monoclonal antibodies are also used in some recipients as induction therapy before and/or after transplantation. Some new immunosuppressive drugs, such as Rapamycin and Everolimus, can not only inhibit the acute transplant rejection but also prevent cardiac vasculopathy. The application of some relatively nontraumatic tests, such as immunological indexes, cardiac markers and other serological parameters, are helpful for diagnosis and preventing postcardiac transplant rejection at early stage and improving the result of cardiac transplantation.
Objective To study the effect of recombinant lentiviral vector mediated human hepatocyte growth factor (hHGF) gene-modified bone marrow mesenchymal stem cells (BMSCs) on the immunological rejection after allograft l iver transplantation in rats, and to reveal the mechanism of immune tolerance. Methods Eight male Sprague Dawley (SD)rats of clean grade (aged 3 to 4 weeks, weighing 75-85 g) were selected for the isolation and culture of BMSCs; 64 adult male SD rats of clean grade (weighing 200-250 g) were used as donors; and 64 adult male Wistar rats of clean grade (weighing 230-280 g) were used as receptors. After establ ishing a stable model of rat allogeneic l iver transplantation, 1 mL sal ine, 2 ×106/mL of BMSCs 1 mL, 2 × 106/mL of BMSCs/green fluorescent protein 1 mL, and 2 × 106/mL of BMSCs/hHGF 1 mL were injected via the portal vein in groups A, B, C, and D respectively. Then the survival time of the rats was observed. The hepatic function was determined and the histological observation of the l iver was performed. The hHGF mRNA expression was detected by RT-PCR, the level of cytokine including hHGF, interleukin 2 (IL-2), IL-4, IL-10, and interferon γ (IFN-γ) by ELISA assay, the level of apoptosis by TUNEL method, and the expression level of prol iferating cell nuclear antigen (PCNA) by immunohistochemical method. Results The survival time of group D was significantly higher than that of groups A, B, and C (P lt; 0.01); the survival time of groups B and C was significantly higher than that of group A (P lt; 0.01), but there was no significant difference between group B and group C (P gt; 0.05). RT-PCR demonstrated the transcription of hHGF mRNA in the grafts of group D; the serum cytokine hHGF reached to (6.2 ± 1.0) ng/mL. Compared with groups B and C, group D exhibited significant inhibitory effect, significantly improved l iver function, and showed mild acute rejection. In addition, the levels of cytokine IL-2 and IFN-γ decreased; the levels of cytokine IL-4 and IL-10 increased; the level of apoptosis reduced; and the expression level of PCNA increased. Except for the expression of IL-4 (P gt; 0.05), there were significant differences in the other indexes between group D and groups B, C (P lt; 0.05). Conclusion BMSCs/hHGF implanting to rat l iver allograft via portal vein can induce immune tolerance. Compared with injection of BMSCs alone, BMSCs/hHGF treatment can alleviate acute rejection and prolong the survival time significantly. The immunosuppressive effect of BMSCs/hHGF is correlated with Th2 shifts up of Th1/Th2 shift, reduced apoptosis, promoted l iver regeneration.
Objective To review the research progress of composite tissue allotransplantation (CTA), analyzethe superiority and the inferiority, and inform the possible direction of further research. Methods Literature concerningCTA was reviewed and analyzed in terms of the l imits of conventional reparative and reconstructive surgery, the definitionof CTA, potential advantages, and treatment risks. Results The cl inical research of CTA both at home and abroad proved that the therapeutic effect of CTA was better than that of conventional reparative and reconstructive surgery. However, therisks resulting from immunosuppressive therapy were still the primary factors restraining the wide cl inical appl ication ofCTA. Conclusion The development of immunosuppressive therapy explores a great development potential for the CTA, and how to decrease the treatment risk of immunosuppressive therapy will be the main research direction in the field of CTA.
Objective To investigate effect of the removal of epithelium and mixed glands from the tracheal allografts on the graftimmunosuppression. Methods Fresh untreated tracheal allografts， cryopreserved tracheal allografts, and 10 off-epithelium tracheal allografts were obtained from 25 male SD rats. Fresh untreated tracheal allografts(40) were divided into 4 groups and dipped respectively in the solution of protease ⅩⅣ in 0, 0.1, 0.3 and 0.5 mg/ml at 4℃ for 12 hours. Thirty recipient male SD rats were randomly and equally divided into group A (fresh untreated tracheal allografts), group B(cryopreserved tracheal allografts), and group C(offepithelium tracheal allografts). The transplanted allografts were implanted into the abdominal cavity of other rats by being embedded in the greater omentum. Twenty-one days after transplantation, the tracheal graft segments were surgically removed, and then were initially fixed in cold 10% neutral buffered formalin solution for hematoxylineosin staining. Histological observation and lymphocyte infiltration were performed on the grafts to evaluate rejection. Results The 0.3 mg/ml protease ⅩⅣ could remove the epithelium and mixed glands of the grafts completely, but did no damage to cartilage. The cartilages of each group all survived and were revascularized. The lumens of group A were filled with granulation and necrosis tissue. In contrast, group B was filled with a few granulation tissues and group C was not at all. The number of lymphocyte infiltration in group A, B, and C was 29.16±2.69/HP, 15.17±2.19/HP, and 11.56±0.87/HP respectively. There was significant difference between group A and both group B and group C (Plt;0.05), and there was significant difference between group B and group C (Plt;0.05). Therefore, the grade of graftrejectionwas group Agt;group Bgt;group C. Conclusion The 0.3 mg/ml protease ⅩⅣ can completely remove the epithelium and mixed glands of grafts at 4℃ for 12 hours, and it preserves the normal structure of cartilage. The antigenicity of tracheal grafts can be greatly reduced by removing the epithelium and by the cryopreservation. The prior tracheal allograft in the omentum is feasible for the revascularization of the grafts.
OBJECTIVE To study the immunosuppressive effect of combined therapy with FK506 and RS-61443 in rat limb allotransplantation. METHODS: A total of 101 male SD rats were randomly divided into seven groups and used as recipients, and 101 Wistar rats were used as donors. All SD rats were performed limb allotransplantation without using immunosuppressants in control group. In experimental groups (Groups 1-6), the recipients were immunosuppressed with various dosages of FK506, RS-61443 or FK506 + RS61443, after transplantation for 5 weeks. To evaluate the results, we observed circulation of the transplanted limb, the mean rejection time, the histologic grading of skin rejection of limb grafts and the survival time of limb grafts. RESULTS: The control group showed rejection signs (edema and erythema of the skin) after a mean time of 3.36 +/- 1.15 days, and the mean survival time of the allografts was only 7.00 +/- 0.78 days. In the groups only using FK506 or RS-61443, the survival time were prolonged to varying degrees, but rejection occurred even in the period of using drug. As dosage increased, the rejection could not be prevented and the damage to liver and kidney could be induced. In the group using FK506 in combination with RS-61443, only skin and muscle of limb allografts showed slight rejection sign, function of liver and kidney was not obviously affected, the mean survival time of limb allografts was prolonged to 58.76 +/- 6.81 days. CONCLUSIONS: A combination of FK506 and RS-61443 is a more potent immunosuppressive agent than FK506 oro RS-61443 in preventing the rejection of limb allografts, and it can obviously prolong the survival time of limb allografts.
Objective To discuss peripheral nerve regeneration under immunosuppression. Methods Current research trends about relationship between peripheral nerve injury and immunoreaction, the experimental result of nerve regeneration after using various immunosuppressors, and the clinical findings after human allogenous hand transplantation were extensively reviewed. Results Peripheral nerve regeneration was accelerated under immunosuppression. Conclusion Peripheral nerve injury may induce immunoreaction, which inhibit nerve regeneration and function recovery.
Objective To investigate effect of intravitreal injection of FK506 on the survival of human retinal pigment epithelial (RPE) cells heter oplastically transplanted into the subretinal space of rabbits.Methods The immortalized human RPE cells were genetically labeled by retrovirus vector carrying a green fluorescent protein (GFP). A total of 50 μl RPE cells suspension with 4×103 cells/μl which expressed GFP were injected into the subretinal space of both eyes of 18 white rabbits and 10 gray rabbits. The left eyes of all of the rabbits were injected of 5 μl FK506 (5 μg/μl) intravitreally once a week during the first 5 weeks, then once every other week until the 20th week and the right eyes were as the control. The histological sections of heteroplastic RPE cells were observed by epifluorescent microscope.Results GFP-expressing cells could be seen after 1 week, 2, 3, 4, 6, 10, 11, 14, 18, 20, 23, 24, 25, 26, 33, and 54 weeks in white rabbits and after 4 , 5, 6, 7, 14, 18, 20, and 26 weeks in gray rabbits. The configuration and integrality of the RPE-GFP cells in the left eyes which had been intravitreally injected of FK506 1-14 weeks after transplantation were better than those in the right eyes without injection. After 18 weeks, the condition of heteroplastic cells with few difference in both eyes in 7 white and 3 gray rabbits were found. After 1-6 weeks, focal and disseminated lymphocytes around the choroidal small vessles of right eyes in 6 white and 3 gray rabbits could be seen while the infiltration of the lymphocytes in the left eyes was much reduced.Conclusion Intravitreal injection of a small amount of FK506 at the first 3 months after transplantation may significantly improve the survival of heteroplastic RPE cells in the subretinal space of rabbits. (Chin J Ocul Fundus Dis，2003，19：333-404)
Objective To find out the beneficial and harmful effectiveness of tacrolimus (TAC) compared with cyclosporine A (CSA) for simultaneous pancreas-kidney transplant (SPKT) recipients. Methods Randomized controlled trials (RCTs) of TAC versus CSA for SPKT recipients were collected from The Cochrane Library, MEDLINE, EMbase, SCI, and CBM database. Bias risk assessment and meta-analysis were performed based on the methods recommended by the Cochrane Collaboration. Results Five RCTs including 342 recipients were included. Pooled data of pancreas survival favored TAC (RR=1.15, 95%CI 1.04 to 1.27; RD=0.11, 95%CI 0.03 to 0.19). However, there were no significant differences of acute rejection (RR=0.81, 95%CI 0.58 to 1.12), patient survival (RR=1.00, 95%CI 0.94 to 1.05), kidney survival (RR=1.02, 95%CI 0.95 to 1.09), and infection (RR=1.00, 95%CI 0.83 to 1.20). Conclusion Based on the recent evidence, TAC results in higher episodes of pancreas survival compared with CSA after SPKT. Treating 100 patients with TAC instead of CSA would increase pancreas survival in 11 recipients.
Objective To investigate the anti-rejection effect and the mechanism of triptolide （TPT） on islet allo- grafts in a murine model. Methods BALB/c mice were used as islet donor. C57BL/6 mice were rendered diabetic by streptozotocin （STZ） injection， and transplanted with islets under the left kidney capsule. The recipients were randomly （method of random digits table） divided into three groups （n=8）. The mice in the treatment groups were injected intrap-eritoneally with TPT at 50 μg/kg （low-dose TPT group， L-TPT group） or 100 μg/kg （high-dose TPT group， H-TPT group） daily in the first 5 days and then on alternate days until 14 days；while the mice in control group were given vehicles （1% tween 80）. Blood glucose after operation were monitored. The grafts were defined as rejection when two consecutive reading of blood glucose＞20 mmol/L. The left kidney of three recipients in each group were resected for pathological examination. The proportion of CD4＋CD25＋Foxp3＋ regulatory T cells in spleen tissues were tested by flow cytometry. Results The median survival time of islet allografts from the control group， L-TPT group， and H-TPT group were 12.6 days （9-16 days）， 21.4 days （14-27 days） ， and 27.6 days （19-34 days）， respectivly. The percentageof CD4＋CD25＋Foxp3＋regulatory T cells in spleen tissues of three groups were （5.2±0.6）%， （12.0±1.3）%， and（15.7±1.8）%， respectivly. Compared with control group， the median survival time of islet transplantation in mice exte-nded and the proportion of CD4＋CD25＋Foxp3＋ regulatory T cells in spleen tissues increased （P＜0.05）. Conclusions TPT could increase the percentage of CD4＋CD25＋Foxp3＋ regulatory T cells， reduce the rejection after islet transplanta-tion， and prolong the survival time of islet transplantation in mice. The immunosuppressive effect of TPT shows a dose-dependent.
ObjectiveTo explore perioperative management model of ABO-incompatible liver transplantation. MethodsThe clinical data of ABO-incompatible caderveric liver transplantions without urgency performed in our center from July 2006 to May 2010 were analyzed retrospectively. Four patients had received an ABO-incompatible graft: AB to O in three, AB to A in one. All the cases were diagnosed as end-stage liver disese, one of them was primary hepatocellular carcinoma. ResultsFour survived to now （11 to 19 months） without severe infections and acute rejections. Two experienced coagulative disturbance and one of them had a second exploration. One developed acute renal failure and recovered with help under continuous veno-venous hemofiltration. All the cases were given 20 mg basiliximab two hours before revascularization and on day 4 after operation respectively. Splenectomy was performed in three, intravenous immunoglobulin was given in all more than seven days. Isohemagglutinin titers were basically stable and not relevant to the clinical manifestations. Antibiotic prophylaxis and immunosuppression protocol was same as the ABO compatible transplants except a 3-month-delay for steroid withdrawal. ConclusionABO-incompatible liver transplantation could be performed with appropriate perioperative management, such as basiliximab induction, splenectomy, intravenous immunoglobulin administration, and routine immunosuppression.