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Purpose To investigate the effect of tolerogenic dendritic cells ( DC) on T lymphocytes in the spleen during the develop-ment of zymosan-induced sepsis in mice, and to explore whether PD-L1 blockade could alleviate the immunosuppressive effect of tolero-genic DC on T lymphocytes. Methods Mice sepsis model was established by intraperitoneal injection of zymosan. Splenic DC and T lymphocytes were isolated respectively by using anti-CD11c and anti-CD3 magnetic beads. The expressions of PD-L1, PD-1 and PIR-B on splenic DC were measured, and IL-12 and IL-10 secreted from DC were determined. Mitogen-induced T lymphocyte proliferation and IL-2 secretion were assessed. Anti- PD-L1 antibody was added into mixed culture of tolerogenic DCs with normal Tcells. T cell proliferation and IL-2, IL-12 and IL-10 concentrations in the supernatant of mixed culture were determined. Results At 5 days and 12 days after zymosan injection, the expressions of PD-L1, PD-1 and PIR-B on splenic DC increased greatly, secretion of IL-12p70 re-duced and that of IL-12p40 and IL-10 augmented in DC, which were associated with decrease of T cells proliferation and IL-2 secre-tion. Administrating anti-PD-L1 antibody into the mixed culture of tolerogenic DC and Tcell could alleviate the suppression of DC on T lymphocyte proliferation and secretion of IL-2, and ameliorate the ability of DC secreting IL-12 and IL-10 as well. Conclusions At late stage of zymosan-induced sepsis, the formation of splenic tolerogenic DC resulted in immunosuppression of T lymphocytes. Anti-PD-L1 antibody could improve the immunoactivity of DC and T lymphocyte through intervening PD-L1/PD-1 pathway.
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BACKGROUND: The tissue engineered corneal epithelium can be constructed with limbal stem cells of healthy rabbit cornea cultured in vitro, and its transplantation may promote the repair and healing of corneal alkaline burn.OBJECTIVE: To probe the effect and opportunity of treating corneal alkaline bum by tissue-engineered corneal epithelium transplantation.DESIGN, TIME AND SETTING: Controlled observational animal experiment was performed in the Department of Experimental Animal at the First Affiliated Hospital of Chinese PLA General Hospital between July 2007 and June 2008.MATERIALS: Twenty-one New Zealand female white rabbits, weighing 2.0-2.5 kg, were divided into control group (n=8, 16 eyes) and transplantation group (n=13, 26 eyes) at random. The transplantation group was further assigned into early transplantation group at 1 day (n=2, 4 eyes), 3 days (n=3, 6 eyes), 6 days (n=6, 12 eyes), and 9 days (n=9, 18 eyes); and mataphase transplantation group at 14 days (n=3, 6 eyes).METHODS: The tissue-engineered corneal epithelium was prepared on 21 rabbits with limbal stem cells cultured in vitro, and reforming comeel alkaline burn models of double eyes were induced by 1 mollL NaOH. The autotogous or allogenic tissue-engineered comeal epithelium transplantation was conducted on early time points (1, 3, 6, 9 days) or metaphase (14 days) after alkaline burning. In the control group, the burns were observed for 4 consecutive weeks.MAIN OUTCOME MEASURES: The ocular surface integrity and histopathology in the transplantation group and control group were observed on several time points within 4 weeks.RESULTS: At 1 weak after corneal alkaline burning, large corneal epithelial dasquamation occurred, 72% incidence of epithelial desquamation or corneal ulcer at 2 weeks, and continued to 4 weeks, while only 25% were observed at 4 weeks in transplantation group, most gained ocular surface integrity; cell infiltration and vascularization of corneal stroma had not bean suppressed in metaphase transplantation group, while they were suppressed obviously in the deep comeal stroma in early transplantation groups; Within 4 weeks, immunological rejection induced by allogenic transplantation was not greater than that of autologous transplantation.CONCLUSION: Cultivated autologous or allogenic tissue-engineered corneal epithelium transplantation could recover ocular surface integrity as soon as possible, and the effect of early transplantation surpasses that of mstaphase.
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Objective:To explore CCR7 expression in splenic dendritic cells and its role in migration and activity of splenic dendritic cells in multiple organ dysfunction syndrome (MODS) in mice.Methods:The MODS model of mice was reproduced by Zymosan injection into peritoneal cavity.The mice were randomly divided into groups of normal,3-6 hours,24-48 hours and 10-12 days post zymosan injection.CD11c and CD205 were analysed by immunohistochemistry;The expression of CD86 and CCR7 of DCs were studied by the flow cytometry analysis.Results:In normal mice,many DC were found at the margin between the red and white pulp.In the 3-6 h and 24-48 h groups,CD86 and CCR7 were strongly up-regulated in the DC,and they distributed in T cells areas.In the 10-12 d group,DC distributed at the margin by the immature form.Conclusion:The CCR7 expression level of DC has close correlations with the migration of DC,CCR7 can be used to evaluate the migration and functional activity of DC in MODS.
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Objective To explore the influence of lung IL-10 on interstitial dendritic cells in multiple organ dysfunction syndrome(MODS),and its role in immunological dysfunction.Methods The model of MODS was replicated by injecting zymosan into the peritoneal cavity of C57BL/6 mice.The mice were then randomly divided into normal groups as well as the groups of 3-6 hours,12-48 hours,5-7 days and 10-12 days post trauma.The level of CCR7 mRNA in lung was detected by RT-PCR and the proteins of CCR7 and IL-10 were immunohistochemically stained.The number of mononuclear cells in MHC-II positive peripheral blood was determined by flow cytometry.Results At the early stage of injury(3-6 hours),the number of IL-10 positive cells and the expression of CCR7 increased,but the number of mononuclear cells in MHC-II positive peripheral blood decreased.At the stage of 12-48 hours,the number of IL-10 positive cells and the expression of CCR7 continued to increase obviously,and that of mononuclear cells in MHC-II positive peripheral blood went on decreasing.At the stage of 10-12 days,the number of IL-10 positive cells increased dramatically compared with that in normal group,but the expression of CCR7 declined obviously in contrast to that in 12-48 hours,and the number of mononuclear cells in MHC-II positive peripheral blood decreased to the lowest level.Conclusions The expression of IL-10 in lung could inhibit the excessive immunological damage induced by dendritic cells on the one hand,and it probably leads to immunosuppression by reducing the CCR7 expression of dendritic cells and the number of mononuclear cells in MHC-II positive peripheral blood on the other hand.
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Objective To investigate the morphology and the distributon of thymus dendritic cells (TDCs) in mouse. Methods TDCs in mouse were observed and analyzed by the electron microscopy and the immunohistochemistry with DCs marker (including CD205, CD1a and MIDC 8). Results There were the most TDCs in the boundary area of thymus cortex and medulla, lesser TDCs in the cortex, and the least in the medulla. TDCs made up to 2 per cent of all cells in thymus. On TDCs surface there were many processes which extended into diastema surrounding lymphocytes. The nucleus was irregular in shape and there were intentations. The cytoplasm was electron lucent and there were few organelles without any lysosme. Conclusion It may effectively count TDCs and display their morphological character that TDC be jointly labeled by the diversiform markers and its ultrastructure observed.
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Objective To determine the dynamic change in IL-12 in MODS mice at different post-trauma time points and its relationship with immunodissonance. Methods 96 male C57BL/6 mice, 6-8 weeks old, were randomly divided into normal control, 3h, 24h, 3d, 7d and 12d post-trauma groups. MODS model were replicated by injection of zymosan into the peritoneal cavity of the mice. Spleen and blood were sampled at different time points. Pathological changes of spleen were observed by light microscope. Immunohistochemistry marker of IL-12p40 in spleen was produced by Powervision method. The levels of IL-12p40 and IL-12p70 in serum and homogenate of spleen were determined by enzyme-link immuno-sorbent assay,and T lymphocyte subsets were analyzed by flow cytometry, respectively. Results Atrophy of the white pulps, proliferation of monocytes and dendritic cells, infiltration of neutrophilic granulocytes in the red pulps could be seen in 12d post trauma group. During the "two hits" in early period (3-24h) and late period (12d) in MODS, the amount of IL-12p40 positive cells increased significantly in the spleen; the level of IL-12p40 increased, whereas the level of IL-12p70 decreased evidently, in serum and the homogenate of the spleen. In the early period, the amount of CD8 + T cells in peripheral blood increased, while the amount of CD4 + T cells reduced obviously in the late stage, thus CD4/CD8 ratio declined in both periods. Conclusions The results indicated that over expression of IL-12p40 by antigen presenting cells in immune organs was correlated with immunosuppression of MODS mice.
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Objective To observe the protective effects of natural antioxidant tea polyphenols(TP) on myocardial free radical metabolic disorder in mice induced by inhalation of hypobaric pure oxygen under 5500 m hypobaric condition. Method Fourty-two male Kunming mice were randomly divided into three groups(n=14 each):group A, normal control; group B, inhalation of pure oxygen(>96%) at simulated altitude of 5500 m in an animal altitude chamber;group C(TP protection group), same as group B but 100 mg/kg of TP was given orally before the exposure. The exposure time was 2 h/d,3 d/wk for a total of 8 wk, and distilled water was given to groups A and B before exposure. After experiment, the mice were decapitated on the next day and the heart was quickly removed. Malondialdehyde(MDA) concentration, superoxide dismutase(SOD) activity and nitric oxide(NO) content were measured. In addition, Cu,Zn-SOD and inducible NO synthase(iNOS) enzymatic contents in myocardial tissue were qualitatively examined by immunohistochemical assaying. Result Compared with the control, MDA concentration, SOD activity and Cu,Zn-SOD enzymatic content in group B were significantly increased(P<0.05).But in TP protection group, myocardial MDA formation was significantly decreased(P<0.01) and SOD activity and Cu,Zn-SOD expression restored to normal. On the contrary, myocardial NO generation and iNOS expression were significantly reduced after repeated inhalation of hypobaric oxygen at 5500 m.NO metabolism regained to normal after repeated administration of TP. Conclusion Natural antioxidant TP had protective effects on myocardial free radical metabolic disorder induced by inhalation of hypobaric pure oxygen under 5500 m hypobaric condition.
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Objective The expression of transforming growth factor beta receptor Ⅰ (TGF?RⅠ) and transforming growth factor beta receptor Ⅱ (TGF?RⅡ) in the synovium and the articular cartilage of patients with osteoarthritis were studied,to explore the possible relationship between the transforming growth factor beta receptor and the pathogenesis and treatment of osteoarthritis.Method The distribution and positive levels of TGF?RⅠ,TGF?RⅡ and TGF? 1 in the synovium and articular cartilage from 26 patients with osteoarthritis and 3 patients with trauma were studied,using Immunohistochemial methods.Results Immunohistochemical staining of TGF?RⅠ showed positive in all synovial samples in patients with osteoarthritis.The positive staining of TGF?RⅠ was found in most synovial lining cells,endotheliocyte and the macrophage in subsynovial layer with osteoarthritis,especially macrophage like synoviocyte.The positive particles were distributed in the cytoplasma.The distribution and staining levels of TGF?RⅡ in synovium and articular cartilage of patients with osteoarthritis were similar to those of TGF?RⅠ.Positive stainings for TGF?RⅠ, TGF?RⅡ and TGF? 1 were found in over half of chondrocytes.Conclusion It is suggested that vehicles of signal trasmission of TGF? are rich in the synovial membrane and cartilage.There is an important effect of TGF? receptor on inhibiting inflammatory process and helping to repair local tissue.
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Objective To analyse the clinical characteristics of primary extra-nodal malignant lymphoma of the ear, nose and pharynx in order to improve the diagnosis of the disease. Methods Clinical characteristics of primary extra-nodal malignant lymphoma of the ear, nose and pharynx were analysed retrospectively in 16 patients. Especially that of the middle ear and paranasal sinuses, which were rare in occurrence. Results There was one patient with lymphoma of the middle ear, presenting otalgia, chronic secretory otitis media (conductive hearing loss) and facial nerve palsy as main symptoms. Seven patients of lymphoma of tonsils showed hypertrophy of the tonsil and foreign body sensation in pharynx as main symptomatology. Two patients of lymphoma in the nose and paranasal sinus complained of bloody rhinorrhea, odorous discharge, and inferior turbinate hypertrophy. The patients suffering from nasopharyx lymphoma always complained of headache and bloody rhinorrhea. Oropharyngeal lymphoma always presented symptoms of pharyngalgia, fever, soft palate ulcer, and ozostomia. Conclusion The keys to make a correct diagnosis of extra-nodal lymphoma of the ear, nose, or pharynx are to become familiar with their clinical characteristics and to perform a prompt biopsy.