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Background Experimental autoimmune uveoretinitis(EAU)is proved to be an organ-specific,T lymphocyte-mediated autoimmune and self-limited disease.Research showed that CD4+CD25+T cell may play important regulation on the course of events,but its mechanism is pending for further study. Objective Present study was to investigate the potential role of CD4+CD25+T cell in the pathogenesis of EAU. Methods Retinal Santigen(S-Ag)was isolated from bovine retinas according tO the procedure as Caspi's previously describe.0.1 ml Santigen(50μg)emulsified with complete Freund'S adjuvant was injected on footpad of 24 inbred adult female Lewis rats aged six to eight-week-old to induce EAU,and 4 normal Lewis rats were as normal control group.Slim-lamp examination was performed to observe the ocular manifestation.Retinal section was prepared in 7,12,15,21 days aher injection for the pathological examination.The pathological grading was on the lnoki'S method.The retinas,inguinal nodes and spleens of rats were obtained in 7,12,15,21 days after injection and the cellular suspension was prepared.Expression of CD4+CD25+T cells on cellular suspension was assayed using flow eytometry.This study complied with the Standard of Association for Research in Vision and Ophthalmology. ResuRs The obvious inflammatory response of the anterior segment was found in S-Ag injected eyes from 7 days through 21 days.The most serious inflammation was found in 12-15 days under the slim-lamp.The hemotoxylin and eosin staining showed the higher pathological grading from 12 to 15 days after injection,showing significant difference in comparison with 7 days and 14 days(P=0.000).In EAU model rats,expressions of CD4+CD25+T cells was significantly increased in retinas, inguinal nodes and spleens in 15 days after injection,showing evidently differences in comparison with control rats(P=0.000). Conclusion The expression level of CD4+CD25+T cells in inflammatory tissue is associated with the inflammation procedure in EAU model rats.This study indicates that CD4+CD25+T cells may play a role in the development of EAU.
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Objective To explore the effect of targeting Kupffer ceHs on CD4~+ CD25~+T cells in schistosome gramtloma.Methods A total of 30 six-to eight-week-old C57BL/6 female mice were divided into three groups equally,namely a control group,an infection group with S.japonicum cercariae(10 cercariae per mouse) and an infection group injected with GdCl_3 through the penile vein(15 mg/kg)twice perweek.After8 weeks of theinfection,the number of CD4~+ CD25~+T cells was detected by using flow cytometry and the number of Foxp3 was detected by using immunohistochemistry.For the detection of murine IL-4,IL-5,IL-10,TGF-β1 and IFN-γ,a DuoSet ELISA development kit was used.Results The number of CD4~+ CD25~+T ceils and the level of IL-10 decreased significantly in the infection group injected with GdCl_3 compared with the infection group.GdCl_3 treatment decreased Foxp3 production and the level of ALT,and reduced the inflammatory response in schistosome Granuloma.Conclusion Kupffer ceils Can regulate the response of CD4~+ CD25~+T cells in schistosome granuloma.
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Objective To investigate whether and how regulatory T cells(Tr) affect macrophages foam-cell formation, and thereby investigate the mechanism of Tr in the development of atherosclerosis. Methods Tr were isolated from lymphocyte suspensions by magnetic cell sorting-column and analyzed by flow cytometry. Macrophages were cultured alone, with CD4~+ CD25~+ T, or CD4~+ CD25~- T cells in the pres-ence of oxLDL for 48 h to transform macrophages into foam cells. Oil red O staining and cellular lipid meas-urement were used to identify macrophage foam cell formation. Semi-quantitative PCR, quantitative real-time PCR and Western blot analysis were carried out to explore the mechanism of Tr-mediated suppression on macrophages foam cell formation. Results Foam cell formation, as identified by oil red O staining, was readily apparent in cells treated with CD4~+ CD25~- T cells and without T cells. After treatment with Tr, a marked decrease(13.9% ± 5.6% ) in foam cell count was observed, compared with untreated cells(13.9% ±5.6% vs 52.9% ± 10.4%, P<0.001 ) or CD4~+ CD25~- T-treated cells(13.9% ± 5.6% vs 53. 1% ± 17.2%, P<0.001 ), 52.9% ± 10.4% and 53.1% ± 17.2%, respectively (P<0.001). The similar effect of Tr was obtained when extracted oil red O and measured by a spectrophotometer. Cholesteryl ester ac-cumulation also used to quantify foam cell formation. Compared with untreated (no T) and CD4~+ CD25~- Tr-treated macrophage cells (CD25~-), the lipids accumulation in CD4~+ CD25~+ Tr-treated macrophage foam cells(CD25~+) was significantly reduced. Total cellular cholesterol and cellular cholesteryl ester was siginifi-cantly reduced in CD25~+ cultures relative to no T[TC(total cholesterol): 57.46 ± 17.92 vs 159.48 ± 16.38, P<0.01 ; CE(esterified cholesterol): 26.68 ± 8.88 vs 102.54 ± 16.67, P<0.001] or CD25~- (TC: 58.50±7.00 vs 150.55±25.11, P<0.01; CE: 26.68±8.88 vs96.90 ± 11.95, P<0.001) cul-tures. Moreover, PCR and Western blot analysis showed that the expression of both CD36 and SRA in Tr-treated macrophage foam cells was significantly down-regulated. Conclusion Results collectively suggest that CD4~+ CD25~+ Tr cells may inhibit macrophage foam-cell formation, which is largely attributed to a down-regulated expression in scavenger receptor in Tr-treated macrophage foam cells.
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Objective To explore the immune function of the whole body and the tumor site of human nasopharyngeal carcinoma (NPC) patients as well as its correlation with CCR4. Methods The ratios of CD4~+CD25~+T cells and CCR4+cells to lymphocytes were measured by flow cytometry in tissues (25 cases) and peripheral blood (35 cases) from nasopharyngeal carcinoma patients, and then statistical analysis was made. Results The ratios of CD4~+CD25~+T cells and CCR4+ cells in tissue and peripheral blood of NPC were higher than those in the control group (P<0.05). In NPC the ratios of the two cells in tissue were higher than in peripheral blood respectively (P<0.05), but there was no such difference between tissue and peripheral blood in the control group (P>0.05). The ratio of CD4~+CD25~+T cells in tissue at stage Ⅲ+Ⅳ was higher than that at stage Ⅰ+Ⅱ of NPC (P<0.05), but there was no such difference between the two stages in peripheral blood in NPC. There was a positive correlation between CD4~+CD25~+T cells and CCR4+ cells in tissue and peripheral blood of NPC (P<0.05). Conclusion NPC patients have different immunosuppression, and there is even more severe immunosuppression in the tumor site. The ratio of CD4~+CD25~+T cells is correlated with the stage of NPC. Therefore, as NPC progresses to later stages, the percentage of CD4~+CD25~+T cells is increased, which is correlated with poor prognosis. CCR4 plays an important role in reactant of CD4~+CD25~+T cells to tumor sites, and is resistant to immunosurveillance.
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Objective To explore the role of Schistosoma japonicum egg antigens in host immune response. Methods Female BALB/c mice aged 6-8 weeks were divided into two groups. The mice in the experiment group were administrated with 10 000 eggs of S.japonicum orally and injected with 200 ?g SEA via tail vein,once for a week. The mice in the control group were infected with PBS. The number of CD4+CD25+T cells was detected in a murine model treated by S. japonicum egg antigens,and the regulatory properties of CD4+CD25+ T cells were assessed while CD4+CD25+ T cells were cocultured with CD4+CD25-T cells. For the detection of murine TGF-? and IL-10,a DuoSet ELISA development kit was used. IL-4,IL-10 and IFN-? were detected by using flow cytometry. Results The number of CD4+CD25+T cells and the level of IL-10 increased in mice treated with S. japonicum egg antigens. CD4+CD25+T cells dramatically enhanced IL-10 production and decreased IFN-? production compared with the CD4+CD25-population. CD4+CD25+T cells suppressed the proliferation of CD4+T cells. Conclusion S. japonicum egg antigens downregulate the host immune response by inducing the production of CD4+CD25+ T cells and IL-10.
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Objective To explore the effect of culture supernatant of Toxoplasma gondii on CD4+CD25+ T cells in mice in vitro. MethodsCD4+CD25+ T cells were separated from spleens of C57BL/6 mice and incubated in the culture superntant of Toxoplasma gondii. The apoptosis percentage of the CD4+CD25+ T cells were detected by FACScan, and the suppression of the CD4+CD25+ T regulatory cells were examined by 3H-TdR incorporation. ResultsCompared with the CD4+CD25+ T cells incubated with RPMI-1640, (36.90?0.36)% CD4+CD25+ T cells took apoptosis after incubated with the culture supernatant of Toxoplasma gondii for 10 hours,and the Annexin-v positive rate increased by (13.60?2.15)%. Compared with RPMI-1640, the culture supernatant of Toxoplasma gondii incubating the CD4+CD25+ T regulatory cells for 5,10 hours reduced their suppressive potential on the proliferation of the CD4+CD25- T cells significantly. ConclusionsSome composition of the culture supernatant of Toxoplasma gondii might cause apoptosis of CD4+CD25+ T cells and as a result, could reduce their suppression on CD4+CD25- T cells.
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Objective:To study the relationship betwen the mechanism of autoimmune disease and CD4+CD25+ T cell population in NIK mutated mice-aly mice.Methods:NIK mutated mice-aly/aly mice were used as model,aly/+mice as NIK normal control;cell populations were determined by FACS and the thymus structure were analyzed by immunohistochemistry.Results:The CD4+CD25+CD8- population were remarkably decreased in aly mice;and the UEA-1 positive cells were absent in aly mice.Conclusion:The autoimmune disease in aly mice might be the result of deceased the CD4+CD25+ population;the UEA-1 positive cells might play an important role in the development of CD4+CD25+ population. [
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Objective:To investigate the expressions of CD4+ CD25+ T cells and IL-10 in the peripheral blood of patients with systemic lupus erythematosus(SLE) and their clinical significance.Methods:Thirty SLE patients(seventeen active and thirteen remissive) and twenty normal controls were enrolled in the study. Flow-cytometric assay was employed for detection of CD4+ CD25+ T cells,and double antibody sandwich ELISA was applied to detect IL-10 in sera from SLE patients and normal controls.Results:The levels of CD4+ T cells in active and remissive SLE were significant lower than normal controls; the positive rate of CD4+ CD25+ T cells in both active and remissive SLE was higher than that in normal controls; the levels of IL-10 in active stage of SLE were significant higher than in remissive stage of SLE or in normal controls. No correlation was found among the levels of CD4+ CD25+ T cells or IL-10 in SLE with SLEDAI scores, anti-DNA and C3 level.Conclusion:CD4+ CD25+ T cells in the peripheral blood of patients with SLE are the marker for activation of T-cells, and aberrant IL-10 production was related with onset of SLE.