To diminish the specific lymphocytes that responsive to the rejection of allograft. Anti-rat CD4,CD8 monoclonal antibodies and trichosnthin (TCS) was conjugated to immunotoxin by heterobifunctional reagent SPDP, 2-IT. The free TCS was removed from conjugates mixture by a column of Sephacryl S-200. The SDS-PAGE and cytotoxic assay was used to measure the biological activity of immunotoxin. SDS-PAGE showed the immunotoxin, free McAb and TCS were in the mixture of conjugation, and the free TCS can be separated by Sephacryl S-200. In Vitro, the lymphocytes of rat can be killed by antiCD4,antiCD8 immunotoxin. The kill capability was relay to the amount of immunotoxin. The authors consider that the immunology unresponsiveness can be induced by antiCD4,antiCD8 immunotoxin. That was useful in induced transplantation tolerance.
Objective To study the effect of allogeneic canine cord blood mesenchymal stem cells（cbMSC）transplantation on the distribution of CD4+T and CD8+T in infracted area of hearts. Methods Mononuclear cells of cord blood were isolated by density gradient centrifugation and amplified by adherent culture. 36 adult male dogs were divided into experimental group and control group. Animal models of acute myocardial infarction were established by ligating anterior descending coronary artery. The fourth generations of mesenchymal stem cells (MSC) were transplanted into infarcted area of hearts by left anterior descending coronary artery after 72h induced by 5-aza and transfected by LacZ. The survival of transplanted cells in hearts can be confirmed by βgal expression. CD4+T and CD8+T cells distributed in infarcted area were detected by immunohistochemical staining method. The ImagePlus 5.1 software was used to analyze the images. Results Cells transplanted into infarcted area could survive for a long time. 2, 4, 8 weeks after transplantation, the IOD of CD4+T in experimental group were 44.35±7.03, 19.29±4.11 and 20.27±3.51 respectively, and the CD4+T/CD8+T ratios were 0.63±0.12, 0.51±0.15 and 0.66±0.08. In control group, the IOD of CD4+T at 2, 4, 8 weeks after transplantation were 65.78±10.27, 28.02±2.59, 29.79±6.83, and the CD4+T/CD8+T ratios were 1.28±0.20, 1.34±0.09 and 1.50±0.16. The IOD of CD4+T and CD4+T/CD8+T ratio in experimental group were significantly lower than that in control group. In experimental group the IOD of CD8+T at 2, 4, 8 weeks after transplantation were 69.88±7.84 , 37.80±8.83 and 30.81±7.42, higher than that in control group which were 51.28±10.01, 20.87±4.50 and 19.91±2.87. Conclusion The preliminary results indicated that allogeneic cbMSC transplanted in infarcted area can escape from immune rejection, its mechanism may be associated withdecreasing the amount of CD4+T cells infiltrated in periphery of infarcted area and maintaining CD4+T/CD8+T ratios at a lower level.
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)
ObjectiveTo assess the prognosis of asymptomatic pulmonary sarcoidosis with normal lung function. MethodsEighty-two patients with asymptomatic pulmonary sarcoidosis,diagnosed in Guangzhou Institute of Respiratory Diseases between July 2007 and November 2013,were followed for 24 months.The clinical data were collected and analyzed. Results31.7% of the patients showed spontaneous remission without therapy,whereas 39.0% deteriorated and 29.3% remain stable.There was no significant difference between the remission group and the progressive sarcoidosis group with regard to FEV1.17.1% of patients in the progressive sarcoidosis group had a decline in FEV1,while 73.8% had a reduced FVC,and 28.3% had a reduced DLCO.At the beginning of observation,TNF-α level was higher in the progressive sarcoidosis group compared with the remission group and the stable group[(173.5±37.8) μg/mL vs.(102.6±82.7) μg/mL and(131.7±57.9) μg/mL,P<0.05].In the progressive sarcoidosis group,TNF-α at the end time of the observation was higher than that at the beginning[(229.2±76.7) μg/mL vs.(173.5±37.8) μg/mL,P<0.05].The neutrophil in BALF was significantly higher in the progressive sarcoidosis group compared with the remission group[(10.6±4.6)% vs.(8.68±5.2)%,P<0.01). There were no significant differences between the progressive sarcoidosis group and the remission group with regard to CD4/CD8 T-cell ratio and the expression of angiotensin converting enzyme(ACE) in lung tissue.Non-necrotizing granulomas rate in endobronchial biopsy was 63.6% in the progressive sarcoidosis group,54.9% in the remission group,and 58.4% in the stable group,respectively,without significant difference between three groups(P>0.05) but was significantly different between three groups at the end time of the observation(69.1% vs.12.7% and 36.2%,P<0.05). ConclusionNearly 40% asymptomatic pulmonary sarcoidosis are likely to deteriorate.The increase of TNF-α and BALF neutrophils may indicate the progress of sarcoidosis.
Objective To investigate the proportions of CD4+ T cells, CD8+ T cells, and mutant of p53 gene in the microenvironment of breast infiltrating ductal carcinoma, and to explore its’ correlation with prognosis of breast infiltrating ductal carcinoma. Methods Eighty-five cases of breast infiltrating ductal carcinoma were collected who underwent surgery in the 371st Central Hospital of Peoples’ Liberation Army from 2010 to 2012, and then detected the proportion of CD4+ T cells and CD8+ T cells, ratio of CD4+ T cells to CD8+ T cells, and mutant of p53 gene in the cancer tissues with immunohistochemistry. Comparison between the sentinel lymph node metastasis group and non-sentinel lymph node metastasis group, mutant of p53 gene group and non-mutant of p53 gene group on the proportions of CD4+ T cells, CD8+ T cells, and ratio of CD4+ T cells to CD8+ T cells were performed, as well as the relationship between proportion of CD8+ T cells/mutant of p53 gene and prognosis of breast infiltrating ductal carcinoma. Results ① The relationship between proportion of CD4+ T cells/proportion of CD8+ T cells/ratio of CD4+ T cells to CD8+ T cells and situation of sentinel lymph node metastasis: at cluster, compared with the sentinel lymph node metastasis group, the proportion of CD8+ T cells was lower in the non-sentinel lymph node metastasis group (P<0.05), but there was no significant difference on the proportion of CD4+ T cells and ratio of CD4+ T cells to CD8+ T cells (P>0.05); at stroma, compared with the sentinel lymph node metastasis group, the proportions of CD4+ T cells and CD8+ T cells were lower, but the ratio of CD4+ T cells to CD8+ T cells was higher in the non-sentinel lymph node metastasis group (P<0.05). ② The relationship between proportion of CD4+ T cells/proportion of CD8+ T cells/ratio of CD4+ T cells to CD8+ T cells and mutant of p53 gene: both at the cluster and stroma, compared with the mutant of p53 gene group, the proportions of CD4+ T cells and CD8+ T cells were lower, but the ratio of CD4+ T cells to CD8+ T cells was higher in the non-mutant of p53 gene group (P<0.05). ③ The relationship between proportion of CD8+ T cells/mutant of p53 gene and prognosis of breast infiltrating ductal carcinoma: the prognosis was worse in patients with high degree of infiltration of CD8+ T cells and mutant of p53 gene than those patients with low degree of infiltration of CD8+ T cells and non-mutant of p53 gene (P<0.05). Conclusions The proportions of CD4+ T cells and CD8+ T cells, and ratio of CD4+ T cells to CD8+ T cells are associated with the situation of sentinel lymph node metastasis and mutant of p53 gene, and the degree of infiltration of CD8+ T cells and mutant of p53 gene are associated with the prognosis of breast infiltrating ductal carcinoma.
ObjectiveTo investigate the role of preoperative peripheral blood CD4/CD8 ratio in predicting the prognosis of patients with coronary atherosclerotic heart disease (CAD) after off-pump coronary artery bypass grafting (OPCABG).MethodsA total of 118 patients with CAD who underwent OPCABG in our hospital from September 2016 to April 2017 were included in the study, including 82 males and 36 females aged 62.74±4.50 years. The primary end point was the incidence of major adverse cardiovascular events (MACE). Patients were divided into a high CD4/CD8 group (≥1.40, 62 patients) and a low CD4/CD8 group (<1.40, 56 patients) according to the results of flow cytometry. The correlation between CD4/CD8 ratio and prognosis of patients after OPCABG and the value of CD4/CD8 ratio for predicting postoperative MACE were evaluated.ResultsMedian duration of follow-up was 23.25 (20.91, 24.70) months, during which 21 patients (17.80%) experienced MACE and 4 patients (3.39%) were lost to follow-up. Kaplan-Meier analysis revealed that high CD4/CD8 group had a significantly higher MACE rate than the low CD4/CD8 group did (log-rank χ2=5.797, P=0.02). The results of adjusted Cox proportional hazards model showed that CD4/CD8 ratio (HR=3.103, 95%CI 1.557-6.187, P<0.01) was an independent risk factor of MACE in patients with CAD after OPCABG. The receiver operating characteristic curve showed that area under curve was 0.778 (95%CI 0.661-0.894, P<0.01), the optimal cut off value was 2.24, the sensitivity was 57.1%, and the specificity was 87.6%.ConclusionPreoperative peripheral blood CD4/CD8 ratio is an independent predictor of MACE after OPCABG in patients with CAD.
Objective To investigate the expression of costimulatory molecules, Fas/FasL, and major histocompatibility complex (MHC)-class II antigens in normal ocular tissues. Methods Twelve eyes were obtained from the eye bank of Zhongshan Ophthalmic Center within 16 to 24 hours postmortem. Six eyes were used for making the retinal wholemounts, and the tissues (iris and ciliary body, choroid, and retina) of the others were used for making the frozen sections. Immunohistochemical staining was carried out on these retinal wholemounts as well as on tissue sections to investigate the exprenion of B7-1 and B7-2 (costimulatory molecules), HLA-DR(MHC class II), CD68 (macrophages), Fas/FasL. Results The results of immunohistochemical staining showed the presence of B7-2, FasL, CD68 and HLA-DR in the iris and ciliary body. Expression of B7-1, FasL, CD 68, and HLA-DR was found in the choroid while HLA-DR, CD68 and FasL were detec table in the retina. Conclusion Expression of costimulatory molecu les, MHC-class II molecules and molecules related to apoptosis is different in the iris, ciliary body, choroid, and retina, which may play an important role in the stability of the immunological microenvironment of these tissues.(Chin J Ocul Fundus Dis，2003，19：109-112)
目的:探讨结直肠癌KAI1/CD82与E－cadherin的表达与预后的关系。方法: 采用免疫组化S-P法检测105例结直肠癌中KAI1/CD82与E-cadherin的表达。结果: 74例中5年生存率为48.6%。生存5年以上的KAI1/CD82的阳性表达率(86.11%),明显高于生存5年以下者(57.89%)(Plt;0.01);而E－cadherin生存5年以上者的阳性表达率(6389%)亦明显高于生存5年以下者(31.58%)(Plt;0.01%)。结论:联合检测KAI1/CD82与E-cadherin的表达对临床判断结直肠癌的预后有重要价值。