【Abstract】ObjectiveTo investigate the effect of xenotransplantation of microencapsulated rabbit parathyroid tissue in different sites in rats for the treatment of hypoparathyroidism. MethodsThe parathyroid glands from Wistar rats were removed to make them aparathyroid. Ultimately, sixteen rats were included because their serum calcium values were continuously below 1.6 mmol/L. We also encapsulated the cultured rabbit parathyroid tissue with alginateBaCl2 microcapsule. According to the transplantation sites, rats were randomly divided into two groups: renal adipose microcapsule group and peritoneal microcapsule group, eight in each group. Encapsulated rabbit parathyroid tissues were then transplanted accordingly to different microcapsule groups. The calcium serum contents were examined on 5，15，25，35，45，55 and 65 d respectively after transplantation and the grafts were observed through electron microscope on the 65 d in particular. ResultsThe calcium contents after transplantation in renal adipose microcapsule group restored to normal and the observation outcomes of grafts showed that they survived well. The calcium contents of posttransplantation in peritoneal group also restored to normal with an exception that it dropped to a level lower than 1.6 mmol/L on the 65 d. Electron microscope also showed that there were necrotic tissues in the center and only a few cells survived on the edge of the grafts. Within peritoneal microcapsule group, the values were significantly lower than others taken at different phases. ConclusionMicroencapsulated rabbit parathyroid tissue that was xenotransplanted into rats can survive and function without administration of immunodepressant. There are significant differences of calcium contents at varying phases between two transplantation sites, which demonstrate that renal adipose may be an optimal site for microcapsule xenotransplantation.
The worldwide shortage in the supply of donor organs and tissues is becoming more pronounced. Xenotransplantation may probably give the hope to overcome the problem ultimately. However, it gives rise to a number of social and ethical issues, among them, the pig appears to be a likely source for human transplantation because it entails least social and ethical issues than no-human primates or other animals and the pig is similar to human in many aspects. The ethical and economic aspects must also be taken into consideration. Patient and his family’s privacy may be stripped because the patient has received a new or unusual treatment. Xenograftings will squint towards a kind of commodities which are different from human graftings and it is a challenge to human transplantation. Xenotransplantation brings a risk of creating new human disease and pandemic, so, it is necessary to formulate a policy and provide input to draft guidelines on the regulation of xenotransplantation.
After escaping from the hyperacute rejection (HAR), the xenograft has to be faced the challenge of acute vascular, acute cellular and even chronic rejection. Endothelial cells have been confirmed as a kind of antigen processing cell (APC) in allo-rejection. The porcine aortic endothelial cell (PAEC) expressed SLA-II and B7 which are the characteristics of professional APC. PAEC also has plenty of alpha-Gal residues, whether the antigen play any role in the post-HAR is still unknown. Human and porcine peripheral blood lymphocyte (PBLC) were isolated and divided into two parts, one for the effectors and the another were incubated with mitomycin C (MMC) as stimulators. The two kinds of PBLC were mixed-cultured within five days. Cultured PAEC from NJZ Pig was incubated with MMC and divided into two: One digested with alpha-galactosidase. The two kinds of PAEC were taken as stimulators to mixed-culture with human PBLC for five days. All the proliferation was detected with 3H-TdR intermingled in the system. The results showed that allo-MLR was ber than xeno-MLR in the cases. The proliferation was much ber when PAEC was used as the stimulator than that of porcine PBLC. However, the response was remarkably decreased after the digestion of alpha-Gal with alpha-galactosidase. The conclusion was that the low response of porcine-to-human MLR in vitro might be related to the predominant indirect pathway of antigen recognition in this system. While PAEC was used as the stimulator the proliferation in MLR was ber which might be concerned that PAEC itself was an APC as well as xeno-antigen sources, thus the direct pathway was predominant and worked more efficiently. The alpha-Gal might induce T cell proliferation through the linkage with the biological big molecules working as a complete antigen. The other post-HAR antigen might also exist in PAEC such as SLA-II, etc.
Insufficient supply of organ for allotransplantation made the study on finding new organ resources from animal progress. Pig is regarded as one of the optimal donor animals for human. The major obstacle in this field is hyperacute reaction (HAR), which is triggered after the xenogenic natural antibodies preexisting in recipient blood combine to the antigens on the surface of the endothelium and activate the complement system. alpha-Galactose residues (alpha-Gal) on the endothelial cell have been identified as the major xenoantigens. NJZ Pig has been closely breed since 1938, whose family history is clear. Tissue samples from heart, liver, kidney, pancreas, lung, small intestine, skin, spleen, thymus and lymph node were obtained and embedded in paraffin. The sections were performed the immunohistochemical staining with the sera from health volunteers (including all the blood types) as the primary antibodies as well as the biotin labeled bandeirae simplicifolia I isolectin B4 (BS I-B4), which has specific affinity to alpha-galactose. All the staining sections were compared with the tissues digested with alpha-galactosidase. There was no difference between the antigens recognized by sera of different blood types. alpha-Gal was still the major xenoantigen on the endothelial cells. There might exist non-alpha-Gal antigens on the distal convoluted tubules and collecting tubules of the kidney. There was no alpha-Gal distributing on the secreting part of pancreas, either the islet cells or the matrix cells, but surely on pancreatic duct and vessels. All the antigenity was destroyed after the enzyme digestion except that the small intestine gland still positive with the BS I-B4. alpha-Gal is the major xenogenic antigen in NJZ Pigs. There exist some unknown antigens on the distal convoluted tubules and collecting ducts of the kidney. The blood type of recipient is not the first affair to be considered in pig-to-human xenotransplantation. The specificity of BS I-B4 for the alpha-galactose needs more detail research.
Objective To review the current condition, test method and progress of the animal model of xeno graft versus host disease(xeno GVHD). Methods The literature review and comprehensive analysis methods were used in this article. Results Implanted immunologic cells, the recepient had the chance of showing host versus graft reaction, GVHD or microchimerism. Now, xeno GVHD could be induced in vivo at small and large animals, it also could be supervised through many ways. Conclusion Chimeric cell is very important to xeno-GVHD animal model. With this model, we can really mimic the immunologic change in vivo after xenotransplantation.
Objective
To evaluate tissue regeneration, body reaction, and biological safety of xenogeneous bladder acellular matrix (BAM) that can be used to repair rabbit bladder.
Methods
Porcine BAM was prepared through physical, chemical, and enzymatic methods, and the effects of acellularization and the structure were observed with HE staining and scanning electron microscope (SEM). Eighteen New Zealand white rabbits (weighing, 2.5-3.0 kg) undergoing partial cystectomy were randomly divided into 2 groups. After partial (about 30%) cystectomy, the porcine BAM was used to replace partial rabbit bladder in the experimental group (n=12), and the incision was directly sutured as control group (n=6). The survival condition of animals was observed after operation. At 15 days, 1, 2, 3, and 6 months after operation, the blood routine, renal function, and electrolyte were tested by collecting the blood samples. At 1, 2, 3, and 6 months after operation, maximum bladder capacity, bladder leak point pressure, and bladder compliance were measured through urodynamic studies. Then gross observation was performed for regeneration of bladder, and the specimens of the bladder were harvested for HE staining and immunohistochemical staining. The surrounding organs and local lymphoid tissues were harvested for gross observation and HE staining.
Results
Cell components were completely removed in the porcine BAM, showing three-dimensional porous structure under SEM. All the animals survived during the experiment. At 15 days after operation, white blood cell count increased, and then returned to normal level in 2 groups, showing no significant difference between 2 groups (P gt; 0.05). The tests of renal function and electrolyte suggested no significant difference between 2 groups (P gt; 0.05). The level of serum creatinine showed a tendency of increase, but it remained within normal range at 6 months after operation. The maximum bladder capacity and compliance in experimental group were significantly higher than those in control group at 3 and 6 months after operation (P lt; 0.05), but no significant difference in bladder leak point pressure at each time point between 2 groups (P gt; 0.05). The urothelial regeneration, smooth muscle regeneration, and blood vessel regeneration were seen by histological observation in 2 groups. In the 2 groups, chronic inflammatory cells infiltration could be observed at 1 month postoperatively, and then chronic inflammatory cells decreased significantly (P lt; 0.05), until complete disappearance. There was no significant difference in score of chronic inflammatory cell infiltration between 2 groups at 3 and 6 months after operation (P gt; 0.05). The α-smooth muscle actin expression was significantly increased with time passing in 2 groups (P lt; 0.05), and it was significantly higher in control group than in experimental group at each time point (P lt; 0.05). In addition, gross and HE staining observations showed no abnormalities in surrounding organs and local lymphoid tissues.
Conclusion
No immune rejection response occurs when porcine BAM is used for xenotransplantation. It is indicated that porcine BAM is relative safety for xenotransplantation.
ObjectiveTo investigate the aim antigen coursing the hyperacute rejection of xenotransplantation. MethodsDocuments about hyperacute rejection in xenotransplantation were reviewed and summarized in detail. ResultsPig is thought to be one of the ideal donors of xenotransplantation, but the major obstacle is hyperacute rejection mediated by complement that is activated though human serum. αGal is recognized as the major antigen and its expression is controlled by α1,3 galactosyltransferase. Immunoabsorption of preexsisted antibody, enzymatic digestion of αGal, knockout αGT gene and transgenic technology have been used to solve this problem. Even so, there remain other antigens which can combine with natural antibodies in human serum, such as, 40×103 molecule in erythrocyte, 210×103, 105×103 and 50×103 antigen in pig embryo brain cell, etc. Conclusion αGal is the major antigen which course the hyperacute rejection. Besides αGal, many nonalphagal need further investigation.
OBJECTIVE: To investigate the role of direct and indirect recognition in pig-to-man xenotransplantation. METHODS: Taken the peripheral blood lymphocytes (PBLC) from three Neijiang pigs and two humans as stimulators and respondors, the one-way mixed lymphatic reactions (MLR) of xenograft were carried out, and allo- and self-PBLC as control. RESULTS: Among the three patterns of MLR, syngeneic was MLR the lowest in proliferation, the allogenic MLR was the highest, and the xenogenic MLR was medium. The PBLCs from humans and pigs were matched on HLA-A, B, DR and DQ by means of modified Terasaki assay. The match on pigs was failure because of the pre-existing natural xenogenic antibody in the testing serum. CONCLUSION: The results suggest that the degree of MHC matching still affect the rejection in xenotransplantation, but the present serum assay of MHC matching is not fit for pig.
OBJECTIVE To investigate the mechanism of xenotransplantation rejection and the interaction between the immunocytes. METHODS: This review concluded the research achievements and new advances in xenotransplantation based on the relevant experimental data. RESULTS: Transgenic pig technology and novel immunosuppressants were applied to suppress hyperacute rejection and acute vascular rejection respectively. Modulation of T cell and antigen presenting cells and induction of tolerance were taken for the prevention of acute cellular rejection. CONCLUSION: In general, the technology of transgenic pig is relatively mature and effective. The mechanism and prevention of acute vascular rejection and acute cellular rejection should be further investigated.
Objective To review the methods of overcoming immunological rejection in xenotransplantation.Methods The strategies of overcoming immunological rejection in xenotransplantation were analyzed and summaried on the basis of an extensive review of the latest l iterature concerned. Results The research development of immunological rejection mechanism and molecular biological technique provided new approaches for overcoming immunological rejection in xenotransplantation. Conclusion It is only a matter of time for xenotransplantation to be appl ied cl inically.
