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Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.
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Limbal stem cells (LSCs) transplantation is an effective surgical treatment to address limbal stem cells deficiency ( LSCD) resulting from Stevens Johnson's syndrome or corneal chemical burn. LSCs transplantation has been reported as the successful stem cell therapy for bodybuilding, while it still faces a number of daunting challenges. Researchers should pay more attention to substitute seed cell, novel cell carrier materials, effective expansion method,niche modification and immune rejection,so that patients with ocular surface diseases would get fullly therapeutic benefit.
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Objective To evaluate the application values of two kinds of single-platform flow cytometric methods,the Volumetric method based on flow sensor and the Trucount method based on Trucount beads,in the counts of T-lymphocyte subsets in peripheral blood of patients after transplantation.Methods The absolute number and percentage of CD4 +,CD8 +,and CD3 + T cells in peripheral blood samples from 107 patients after liver or renal transplantation were determined by the Trucount method and the Volumetric method,respectively,and their results were compared using paired t-test and linear regression analysis.Five samples with low CD3 + counts were selected and the precisions of the absolute number of CD4 +,CD8 + and CD3 + T ceils detected by the Volumetric method were evaluated.Results There was no significant difference in the levels of CD4+,CD4+/CD3+,CD8+,CD8+/CD3+,and CD4+/CD8 + in peripheral blood between the Trucount method and the Volumetric method (P ＞ 0.05),and the linear regression coefficients between them were from 0.9 to 1.1.When the concentration of CD3 + was equal or more than 40/μL,the coefficients of variation (CVs) were below 5.5％ for the Volumetric method.When the concentration of CD3 + was 20/μL,the CVs of CD3 +,CD4 +,and CD8 + were 5.19％,10.28％ and 6.48％,respectively.Conclusion The single-platform method based on flow sensor is accurate and reproducible for counting T-lymphocyte subsets in peripheral blood,which may be used to monitor the immune state of the patients after liver or renal transplantation.
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Aim: To evaluate the therapeutic effect of subconjunctival Bevacizumabon corneal neovascularisation Design: A prospective randomized noncomparative study. Method: The charts of 10 consecutive patients with corneal neovascularisation who received single S.C. inj. of Bevacizumab (2.5mg/0.1ml) were reviewed. Digital photographs of the cornea were taken pre & post injection & then at 1 wk,3wk & 2months duration. Digital photographs of the cornea were analyzed to determine the length, density, extent, centricity of corneal neovascularisation and the area of cornea covered by neovascularisation as a percentage of the total corneal area. Results: Subconjunctival injection of Bevacizumab (Avastin) caused significant regression of corneal neovascularisation in 1 pt, partial regression in 6 pts and no effect in 3 pts as measured by length and surface area of neovascularisation. No significant ocular or systemic complications were found. Conclusion: Subconjunctival inj. of Bevacizumab is effective in regressing corneal neovascularisation.
Subject(s)
Antibodies, Monoclonal, Humanized/administration & dosage , Conjunctivitis, Acute Hemorrhagic/drug therapy , Corneal Neovascularization/drug therapy , Humans , Injections, Intraocular , Sodium Glutamate/administration & dosageABSTRACT
· AIM: To evaluate the combined effect of rapamycin ophthalmic solution and cyclosporin A ophthalmic solution on allogeneic transplantation in a rat model.rats were used as donors. The animals were randomly assigned to 4 groups. Group A: control (ophthalmic solution solvent 100μ L); group B: 2g/L rapamycin ophthalmic solution 100μ L; group C: 10g/L cyclosporin A ophthalmic solution 100μ L; group D: 2g/L rapamycin ophthalmic solution 50μ L and 10g/L cyclosporin A ophthalmic solution 50μ L, 4 times every day. The treatment was started on 2d after operation, and the animals were administered until rejection. The grafts were inspected by slit-lamp microscope and the corneal survival time was recorded. The pathologic changes were measured by light microscope.significantly prolonged compared with the control group (P ＜ 0.01). However, the combined therapy (group D) was significantly superior compared with group B and C (P＜0.05, P＜ 0.01). The histopathological findings showed that the inflammation cells, neovascularity in each treated group were significantly fewer than that in control group at 14d after operation.a double drug regimen with rapamycin ophthalmic solution and cyclosporin A ophthalmic solution for the control of acute corneal allograft rejection. It indicats that the combined therapy produced synergistic effect.
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Objective To study the histocompatibility of tissue engineered skin with the observation of the effects of selectins E and P on the immunological rejection after skin allograft in rats. Methods Tissue engineered skin was prepared as follows: The materials obtained from the neonatal SD rats were cultured and then grafted onto the adult Wistar rats. The expression of selectins E and P in the grafted skin was determined with immunohistochemical staining. Results The expression of selectins E and P was significantly higher in the rats with allograft than in the rats with the grafts of tissue-engineered material. Conclusion Selectins E and P play an important role in the immunological rejection after allograft of skin but the tissue-engineered skin graft possesses favorable histocompatibility and shows no obvious immunological rejection.
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AIM: To observe the effect of cornus of ficinalis glycosides(COG) ophthalmic solution on the corneal allograft rejection by topical instillation. METHODS: The corneal transplantation model on the closed colony rats was established. The rejection time of all animals was recorded and compared by slit-lamp microscope. The pathologic changes were measured by immunohistochemistry and scanning electron microscope.RESULTS: The histopathological and immunohistochemistry findings showed that the lymphocytes, neovascularity and the expression of ICAM-1 in COG-treated group were significantly fewer than that in control group at 15 d after operation.CONCLUSION: COG ophthalmic solution prevents and suppresses the corneal allograft rejection.