Objective To study degradation of the antigen-extracted meniscus in PBS solution with no enzyme or with different enzymes. Methods Four types of enzymes (collagenase, hyaluronidase, trypsin, papain) were used to enzymolyze the antigen-extracted meniscus and the fresh meniscus for 3, 7, 15 and 30 days (37℃). The antigenextracted meniscus and the fresh meniscus were immersed in PBS solution (37℃) for 30 days. Weight loss measurement, UV spectrophotometry, and scanning electron microscopy (SEM) were used to characterize the degraded materials. Results The two types of the materials were remarkably digested under the enzymes, especially under trypsin. The degradation curves showed that the antigen-extracted meniscus was enzymolyzed less than the fresh meniscus. The degradation products were grouped as amino, peptide, and polyose by the analysis. Both of the materials could hardly behydrolyzed in PBS solution without the enzymes. The four different enzymes had different surface morphologies under the examination of SEM. Conclusion The antigen-extracted meniscus is enzymolyzed more slowly than the fresh meniscus in vitro, and the result can be used as a guideline to the further research.
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.
Purpose To investigate whether experimental autoimmune uveitis can be induced equally in different rats by urea soluble fraction of bovine melanin-associated antigen(USF-BMAA),and,if so,difference among them. Methods Lewis rats,F344 rats,Wistar rats were immunized with USF-BMAA emulsified with complete Freud is adijuvant and Bordelella pertussis to induce experimental autoimmune uveitis.The animal models were investigated clinically and histopathologically and compared with each other. Rusults Experimental autoimmune uveitis could be induced in Lewis rats,F344 rats and Wistar rats with US-BMAA.Clinical and histopathalogical examination showed that bilateral ocular inflammation developed after immunization 9-13 days.Although inflammation was mainly located in anterior uvea,a mild focal choroiditis was noted in those with severe anterior inflammation.No inflammation was observed in the retina and pineal gland.Experimental autoimmune uveieis induced with USF-BMAA was similar to experimental autoimmune anterior uveitis incited with BMAA presented by other authors.Inflammation induced with USF-BMAA in F344 rats and in Lewis rats was quite similar in the severity and course of the model.But the inflammation was less in Wistar rats compared with that in Lewis rats and F344 rats. Conclusion Experimental autoimmune anterior uveitis was successfully induced with USF-BMAA in Lewis rats,F344 rats and Wistar rats.The difference with regard to the severity among these aminals were propably attributed to their genetic bankground. (Chin J Ocul Fundus Dis,1998,14:149-152)
Objective To investigate the correlation of expression of Fas/Fas ligand (FasL) and apoptosis in retinoblastoma (RB)． Methods The expression and distribution of Fas/FasL were detected by using immunohistochemical staining in 32 cases of RB． Light microsc opy (32 cases), electron microscopy (4 cases) and TdT mediated biotin-d UTP nick-end labeling (TUNEL) (12 cases) were used to study apoptosis in RB． Results Apoptotic RB cells mostly located at RB regress area． Chromatin margination and apoptotic bodies were found in RB． TUNEL posi tive labeling cells especially located in tumor regress area． Positive immunola beling for Fas and FasL was found in all RB specimens． There was a highly signi ficant and positive correlation between the expression of Fas/FasL and apoptotic indices (AI) (Plt;0.01 or 0.001)． Conclusion The results suggest that apoptotic cell death is prevalent in RB and it may be one type of the most dominant cell death． Fas system may play an important role in oncogenesis and progression of RB, and the up-regulation of Fas system expression might induce RB cell apoptosis． (Chin J Ocul Fundus Dis, 2001,17:21-23)
Regulatory T cells (Treg) are critical for regulation of tolerance, control immune responses to self-antigens thereby preventing autoimmunity, and limiting responses to foreign antigens thereby minimizing T cell-mediated immunopathology. Recent data indicate that suppression of organ-specific autoimmunity is dependent on the antigen specificity of Treg. An emerging model of Treg action is that organ-specific Treg acquire suppressive activity through activation by dendritic cells expressing specific antigens. Thus, the efficacy of Treg-based therapy should be increased by using antigen-specific Treg rather than polyclonal Treg. It is necessary to identify relevant antigens and to expand antigen-specific Treg from polyclonal populations. Here, we discuss recent techniques for expansion of antigen-specific Treg, function and antigen specificity of Treg and the therapeutic potential of Treg in controlling autoimmune disease and inducing transplant tolerance.
Objective To evaluate which is better method zymogen or low temperature frozen in removing vascular endothelial cell so as to lay a foundation for creating a kind of brace which is not to be rejected and the same as own blood vessel. Methods Fresh and not damaged umbilical blood vessel was collected from natural labour women, human umbilical blood vessel was remove carefully from normal foetus, then was put into disinfectant at 37℃ for 24 hours. They were divided into 3 groups:normal group(NG),zymogen group(ZG) and low temperature frozen group(LG). ZG: 0.1% collagenⅡ enzyme was addedin umbilical blood vessel and closed the both sides and the vascular endothelialcell was removed in 37℃ water. LG:Umbilical blood vessel was put into liquidnitrogen for 24 hours after frozened step by step, and then it was put into 37℃ water for 30-60 s and the vascular endothelial cells were washed away by normal saline. NG:Umbilical blood vessel was kept into 4℃ Kerb’s liquid. The bacteria were culturedin each group. The samples were stained by HE,elastic fiber and collagen fiberwere observed by light and scanning electron microscope. The difference of compliance was compared. Human leukocyte antigen ABC(HLA-ABC) and HLA-DR were observed by immunohistochemical method and the expression of antigen of umbilical blood vessel was analysed. Results In LG, umbilical vascular endothelial cells were removed completely; artery showed vertical smooth muscle and vein showed elastic membrane. InZG, umbilical vascular endothelial cells were removed completely after 20 minutes;artery showed vertical smooth muscle cells and vein showed lower endothelial layer. The vascular compliance in LG was higher than that in NG, and the latter was also higher than that in ZG,but showing no significant differences (Pgt;0.05). The compliance of umbilical vein was 2-3 times as much asthat of umbilical artery.The expression of HLA-ABC and HLA-DR in LG andZG were lower than that in NG, showing significant differences (Plt;0.01). Conclusion Low temperature frozen methodand zymogen method(0.1% collagen Ⅱ enzyme for 20 min) can remove vascular endothelial cells of human umbilical blood vessel completely.Low temperature frozenmethod was better than zymogen method.
The expression of T antigen in rectal cancer and mucosa remote from carcinoma by immunohistochemistry was investigated. Mucin protein was also examined by HID-AB staining. The results showed that the expression of T antigen in rectal cancer was much ber than those in 10cm mucosa remote from carcinoma and no significant difference as compared with 5cm mucosa. The sialomucin reactions in 5cm and 10cm mucosa remote from carcinoma were 45% and 20% respectively. The coincident sialomucin positive reaction and expression of T antigen were found in 40% 5cm remote mucosa .There is significant correlation between them (Ｐ＜0.05). The authors conclude that the expression of tumorrelated antigen and change of mucin protein in remote mucosa without malignant invasion may suggest the malignant potential of the mucosa. Further investigations should be performed into the effect of these changes on the local recurrence after redical resection of rectal cancer.
PURPOSE: To investigate the activation and immune respones of lymphocytes in epiretinal membranes (ERMs)and subretinaI membranes (SRMs). METHODS: A panel of morioclonal antibodies against CD23 (activated B cell), CD25 (activated T cell), CD68(macrophages) and HLA-DR (human leukocyte antigen II antigen)were used for the study of 20 specimens of ERMs from 20 patients with proliferative vitreoretinopathy (PVR),traumatic PVR and secondary traction retinal detachment,and 2 SRMs from PVR and traumatic PVR, with positive and negative reaction specimens as controls. RESULTS:Four cases of ERMs were found to be CD23 and CD25 positive respectively,and one case of SRMs to be CD23 and CD25 positive respectively. All the specimens of ERMs and SRMs revealed CD68 and HLA-DR positive in this series. CONCLUSIONS :There might be an aberrant immunoreaction mediated by T and B cells in the ERMs and the SRMs,and they might play an important role in the patbogenesis of PVR,traumatic PVR and secondary traction retinal detachments. （Chin J Ocul Fundus Dis,1996,12: 147-150）
Objective To investigate the effects of gamma;-interferon (IFN-gamma; on the expression of costimulatory molecules B7-1 (CD80) and B7-2 (CD86) and Fas/FasL in human retina. Methods Nine human eyes were obtained from the eye-bank of Zhongshan Ophthalmic Center. Six eyes were used for making retinal wholemounts, and 12 retinal wholemounts from each eye were put into the 24-hole culture board which had 2ml DMEM/F12 culture medium in each hole. The wholemounts were divided into 3 groups whose concentration of IFN-gamma; was 0, 200, and 1000 U/ml respectively. After cultured in 37℃ culture box (95%O 2，5%CO 2) for 24 hours, the expressions of B7-1 (CD80), B7-2 (CD86), and Fas/FasL on these retinal wholemounts were detected by immunohistochemical method. The retinal wholemounts from 3 healthy people were detected by immunohistochemical method as the control. Results Expression of FasL but not B7-1, B7-2 or Fas was found in the control group, while the expression of B7-1, B7-2 and Fas and increased expression of FasL were found after cultured with IFN-gamma;. Conclusion IFN-gamma; may be involved in the occurrence of ocular immune response and induction of apoptosis via the stimulation of expression of costimulatory molecules and Fas/FasL, which plays an important role in the activation of T lymphocytes. (Chin J Ocul Fundus Dis, 2006, 22: 117-119)
Objective To observe the influence of the expression of CD18 on the neutrophile and the leukocyte adhesion to retinal vascular endothelium by hypoxia-inducible factor-1 alpha (HIF-1alpha;) in early diabetic retinopathy rats. Methods Male Sprague-Dawley rats received intraperitoneal injection of streptozotocin to induce diabetes model. 18 diabetic rats were divided into 3 groups randomly after 2 months of diabetes induction, including diabetic group (group B), HIF-1alpha; anti-sense oligonucleotides (ASODN) injection group (group C) and HIF-1alpha; sense oligonucleotides (SODN) injection group (group D), the age and weigh matched health rats were chosen as control group (group A), with 6 rats in each group. Then group A and B rats received 5% glucose solution caudalis veins injection, group C and group D rats received HIF-1alpha; ASODN and HIF-1alpha; SODN caudalis veins injection, respectively(025 mg/kg).The level of CD18 on the neutrophil isolated from the peripheral blood was measured by flow cytometry. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Results The percentage of CD18 positive neutrophil cell was（44.93plusmn;3.60）% in group B，（18.66plusmn;1.52）% in group A，（31.66plusmn;4.72）% in group C，（51.00plusmn;5.66）% in group D. Compared with each other groups，the differences are statistically significant （F=42.46, Plt;0.001）. The number of positive staining cells of retinal leukocyte was （46.16plusmn;10.68）in group A，（133.83plusmn;20.43）in group B,（99.83plusmn;9.28）in group C，（121.33plusmn;10.23） in group C. Compared group B with group C，the number of positive staining cells raised about 2.89 times；compared group B with group C and D，the differences are statistically significant （P=0.12，95% confidence interval -3.69~28.69）. Conclusions In vivo, HIF-1alpha; can decreased the expression of CD18 on neutrophils from diabetic ratsprime; peripheral blood and the collection of retinal leukostasis in the diabetic animals. HIF-1alpha; may serve as a therapeutic target for the treatment and/or prevention of early diabetic retinopathy. （Chin J Ocul Fundus Dis，2008，24：268-271）