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Background Researches demonstrated that CpG ODN,a immunostimulatory sequences,has preventing and treating effect on allergic conjunctivitis caused by protein allergen.However,its effect on allergic conjunctivitis caused by fungal allergen is unclear. Objective This study aimed to investigate into whether the Th1-Th2 switching immunostimulatory CpG ODN could reverse the response in the murine allergic conjunctivitis model caused by aspergillus fumigatus. Methods A mixture of spores and hyphae of aspergillus fumigatus strain was used to induce allergic conjunctivitis in male BALB/C mice aged 6-8 weeks.This experiment was designed into preventive or therapeutical treatment program.Under both settings,allergic conjunctivitis of the animals were treated with CpG ODN,nonstimulatory GpC ODN or PBS.After the last challenge with the allergen,the clinical symptoms of the animals were scored based on the criteria of Magone.The animals were sacrificed and the histopathological examination of conjunctiva was performed.Expression of TLR4 mRNA in conjunctiva was analyzed by real-time PCR assay.The responsiveness and populations of lymphocytes in spleen and draining lymph nodes were analyzed by flow cytometry.The use complied with the Standard of Association for Research in Vision and Ophthalmology. Results In the prevention mode.CpG ODN decreased subconjunctival infiltration compared with GpC ODN and PBS groups with the average neutrophil count index(21.25 ±11.59/section,30.75 ±11.44 section and 69.00±9.90/section,respectively).Expression of TLR4 mRNA was up-regulated significantly by CpG ODN.The clinical scores for CpG ODN group were insignificantly lower than those in GpC ODN group and PBS group(P＞0.05).In the therapeutic mode,compared with GpC ODN and PBS groups,the allergic symptom score in CpG ODN group manifested significantly lower(t=4.000.t=2.750,P＜0.01)and showed fewer cellular infiltration(t=4.870,t=3.829,P＜0.01)and higher expression of TLR4 mRNA(P＜0.01).In cultured splenic and draining lymph node cells,increased percentages of CD4+ CD25+ and CD4+ CD25+ CD69+ in CpG ODN group were observed compared with control groups(|P＜0.05). Conclusion CpG ODN can relieve aspergillus fumigatus-induced allergic conjunctivitis via either subconjunctival injection or topical application by upregulating expression of TLR4 and activating Treg lymphocytes.
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Objective To investigate the effeet of eyclosporine A (CsA) on CD4+ Foxp3+ T cells in C57BL/6 mice and the underlying mechanism.Methods C57BL/6 mice, aging 8～ 10 weeks, were administered intraperitoneally with CsA at a dose of 20 mg· kg-1·d-1 for 2 weeks, while the control mice given sterile PBS.After 2 weeks, peripheral blood was collected, and splenoeytes and thymoeytes were prepared for the detection of CD4 + Foxp3 + T cells by using flow cytometry.Results The proportion of CD4+ Foxp3+ T cells in peripheral blood, spleen and thymus of CsA-treated mice was (0.581 ± 0.089) %, (2.189 ± 0.046) % and (0.472 ± 0.049) %, while that in PBS-treated mice was (1.751 ± 0.227) %, (3.684 ± 0.169) % and (1.412 ± 0.188) %, respectively (P < 0.001).Conclusion The reduction of CD4 + Foxp3+ T cells in CsA-treated mice suggests that CsA can inhibit the development of regulatory T cells.
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CD4~+T helper cells not only activate CD8~+T cells,but also play a pivotal role in the gener- ation and maintenance of memorial cytotoxic T lymphocytes(CTL).In addition, it is shown that CD4~+T cells have direct anti-tumor function.However,CD4~+ CD25~+ regulatory T cells(Tregs) negatively control immune re-sponse,and play an important role in immunosuppression and immune escape, it is considered as a key reason for ineffective immunotherapy.In recent years,there is great progress in cancer immunotherapy,and relevant cancer vaccines have attracted a wide spread attention.
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Objective To study the correlation between the percentage of intrahepatic regulatory T cell (Treg) and liver inflammatory activity in chronic hepatitis B (CHB) patients.Methods Twenty-six cases of CHB patients admitted to First Department of Internal Medicine,Nagasaki University School of Medicine,Sakamoto,Nagasaki,Japan were enrolled and performed liver biopsy in this study.CD3+cell and Foxp3+ cell in liver were detected by immunohistochemistry.the percentage of Foxp3+/CD3+cell was determined.Clinical data including alanine aminotransferase (ALT).aspartate aminotransferase(AST), hepatitis B virus (HBV) DNA level and the inflammatory activity of histological activity index score (HAD of liver pathology using Knodell evaluating system were collected.The data was analyzed by SPSS 13.0 software.Results Foxp3+Tregs in serial sections of CHB accumulated mainly in the portal area.There Was a significance correlation between the percentage of Foxp3+/CD3+ cell and liver parenchyma inflammation (P=0.007 6).Moreover,Foxp3+Treg in CHB patients with high serum ALT or AsT level presented in a higher frequency than in patients with low AIT or AST level.The difference between these tWO groups was statistically significant (rALT=O.438,PALT=0.025; fAST=O.436,Past=O.026).There Was a tendency between the percentage of Foxp3+/CD3+ cell in liver and HBV DNA level.however the correlation was not statistically significant.Conclusion It is suggested that CD4+ CD25+ Foxp3+Treg may play a major role in the pathogenesis of liver injury in the CHB patients.
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Objective To investigate the therapeutic effect of 4-1BB monoclone antibodies on mice hepatitis induced by Coneanavalin A (ConA) and its influenes on CD4+CD25+T lymphoeytes during the course. Methods The miee model of hepatic injury was indueed by ConA and monitored by hepatic function tests and hepatic pathology. The expressions of 4-1BB were examined by flow eytometry. 4-1BB monoelone antibodies were intravenously injected to the mice. The therapeutic efficacy was then examined by hepatic function tests and hepatic pathology. The expressions of CD4+CD25+T lymphoeytes were also examined by flow eytometry. Results The group of immune hepatic injury induced by ConA showed damage and marked increase of ALT and AST which were (139±22) U/L and (130±16) U/L respectively. The expression level of 4-1BB was 8.1±2.6. Compared with the eontrol group, the difference was significant (P<0.05). The overall eondition of the miee was improved after being treated with 4-1BB monoelone antibodies. ALT and AST were lowed down to (98±14) U/L and (89±11) U/L respectively and the differenee was signifieant (P<0.01). The expression of 4-1BB of the control group was 3.0±0.8 and that of the treatment group was 8.3±3.0. The difference was significant (P<0.01). Conclusion 4-1BB eontributes to the immune-mediated hepatic injury induced by Con-A.
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Using indirect immunofluorescent technique and immunohistochemical staining method,we detected 14 patients with progressive Mooren's ulcer for T lymphocyte suhpopulations(Pan-T,Th,Ts)and the ratio of Th/Ts in peripheral blood as well as in the limbus conjunctiva near the corneal ulcer.It was found that Pan-T,Th,Ts and Th/Ts ratio in con- junctiva were significantly higher than that in normal control,whereas these parameters in peripheral blood showed no differences from that in the control group.From the above data,we may conclude that the intensive immune activities due to the increased local Th/Ts ratio play an important role in the process of Mooren's ulcer.
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The changes of T cell subpopulations of peripheral blood were studied in children with acute leukemia by means of McAb-SACI-Ig rosette mcthod. The results show that OKT8~+ cells in patients with acute lymphocytic leukemia (ALL) and aplastic anaemia (AA) increase and the ratio of OKT4~+ to OKT8~+ decreases significantly, but there is no significant change of T cell subpopulations in the cases with acute granulocytic leukemia. It suggested that inhibitory T cell is related to the development of ALL and AA.
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Objective:To observe the change of immunity in vivo after Vitreous cryopreservation with tetramethylpyrazine nerve allograft was implanted into the recipient so as to judge its immunogenicity.Methods:Sciatic nerves of Sprague-Dawley rats were resected and made by Vitreous cryopreservation with tetramethylpyrazin.The nerves from Sprague-Dawley rats were implanted into the dorsal and ventral subcutis of Wistar rats in experimental group。Fresh nerves from Sprague-Dawley rats were implanted into the dorsal and ventral subcutis of Sprague-Dawley rats in control group respectively.T lymphoeytes infiltraion in isograft nerve was investigated by immunohis-tochemical Methods:staining CD4 and CD8 positive T lymphocytes with anti-CD4 and CD8 monoclonal antibodies(MoAb).Results:320 mg/L tetramethylpyrazine combined vitreous cryopreservation stainings at-20℃for 3 weeks showed that the number of CD4 and CD8 positive T lymphocytes infiltrated into the nerve implants of the experimental group was much lower than that of other tetramethylpyrazine combined vitreous cryopreservation in the control group.there was no marked difference of the 320mg/L tetramethylpyrazine combined vitreous cryopreservation stainings at-20℃for 3 weeks nerves group compared with autografting group.Conclusion:The nerve treated by 320 mg/L tetramethylpyrazine combined vitreous cryopreservation at-20℃for 3 weeks shoued lower immunogenicity after implanta-tion,and there was no significant rejection response after transplantation in rats.