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Regulatory T (Treg) cells is an indispensable subset of T lymphocyte with the ability of immunosuppression in the periphery.Thymus-derived Treg cells(CD4+CD25+FOXP3+Treg cells) play a fundamental role in maintaining immune homeostasis in vivo.Treg cells have been actively involved in the onset and development of major human diseases including malignant tumors, autoimmune diseases,infectious diseases,allergic diseases and graft versus host disease(GVHD).Exosomes are membranous vesicles of endosomal origin that released from multiple cells into the extracellular space.They are currently considered to be vehicles containing protein,RNA and MicroRNA which can been transferred to recipient cells to modulate their activity by cell-contact-independent mecha-nism.Exosomes have a great impact on the induction and proliferation of Treg cells,understanding the relation of them will lead to novel therapeutic approaches for cancer immunotherapy,treating autoimmunity,infection,allergy and organ transplantation.
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Objective To compare the synchronous changes of high risk human papillomavirus load(HPV-DNA)and CD4+CD25+Foxp3+regulatory T cells(Treg)in local microenvironment of cervix,and investigate the ef-fects of HPV virus replication and progression of cervical lesions on Treg cells. Methods 304 cases of HR-HPV infection with cervical lesions were divided into 5 groups,cervical intraepithelial neoplasia(CIN)I,CINII,CINIII, cervical cancer and chronic cervicitis.The HPV-DNA of cervical secretion was detected by PCR fluorescence,and the relative Treg cells numbers from cervical brush samples were determined by flow cytometry with CD4+CD25+Foxp3+gating,and the data were statistically analyzed. Results(1)There was significant difference of cervical Treg cells in different degrees of cervical lesion and different copy numbers by variance comparison(F = 24.93, 109.86,P < 0.05),and a further pairwise comparison showed that there was no significant difference of Treg cell between chronic cervicitis and CINI and low load(HPV DNA 104~105copies/mL)and medium load(HPV DNA 105~106copies/mL)(P>0.05).There was a significant difference between the other groups(P<0.05);(2)Treg cells as variable,interaction effect of cervical lesions and viral load factors were significant different(F=3.39,P<0.05). The effect of different cervical lesions and HR-HPV viral load on the expression of Treg cells was differen-tial. An overall showing with the degree of cervical lesions increased,HR-HPV virus copy number increased, Treg cells expression increased gradually;(3)CD4+CD25+Foxp3+Treg cells were highly expressed in cervical cancer patients,but the expression level fluctuated widely and the numerical distribution was the most dispersedly. Conclusion The immune suppression function of local CD4+CD25+Foxp3+Treg cells with different cervical lesions and different HR-HPV DNA may bilaterally regulate the prognosis of cervical lesions,as a whole,between Treg cells and HR-HPV load and cervical lesions were the consistent progress trend.
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Japanese encephalitis (JE) is neuroinflammation characterized by uncontrolled infiltration of peripheral leukocytes into the central nervous system (CNS). We previously demonstrated exacerbation of JE following CD11c(hi) dendritic cell (DC) ablation in CD11c-DTR transgenic mice. Moreover, CD11c(hi) DC ablation led to abnormal differentiation of CD11b⁺Ly-6C(hi) monocytes and enhanced permeability of the blood-brain barrier (BBB), resulting in promoting the progression of JE. Here, we examined changes in lymphoid and myeloid-derived leukocyte subpopulations associated with pro- and anti-inflammation during JE progression. The analyses of this study focused on regulatory CD4⁺Foxp3⁺ regulatory T cells (Tregs), IL-17⁺CD4⁺ Th17 cells, and CD11b⁺Ly-6C(hi) and Ly-6C(lo) monocytes. CD11c(hi) DC ablation resulted in the accumulation of IL-17⁺CD4⁺ Th17 cells in the CNS, thereby leading to lower ratio of Tregs to Th17 cells. This result was corroborated by the higher expression levels of IL-17 and RORγT in CD4⁺ T cells from the brains of CD11c(hi) DC-ablated mice. In addition, CD11c(hi) DC-ablated mice showed higher frequency and total number of inflammatory CD11b⁺Ly-6C(hi) monocytes, whereas CD11b⁺Ly-6C(lo) monocytes were detected with lower frequency and total number in CD11c(hi) DC-ablated mice. Furthermore, CD11c(hi) DC ablation altered the phenotype and function of CD11b⁺Ly-6C(lo) monocytes, resulting in lower levels of activation marker and anti-inflammatory cytokine (IL-10 and TGF-β) expression. Collectively, these results indicate that CD11c(hi) DC ablation caused an imbalance in CD4⁺ Th17/Treg cells and CD11b⁺Ly-6C(hi)/Ly-6C(lo) monocytes in the lymphoid tissue and CNS during JE progression. This imbalanced orchestration of pro- and anti-inflammatory leukocytes following CD11c(hi) DC ablation may contribute to the exacerbation of JE.
Subject(s)
Animals , Humans , Mice , Asian People , Blood-Brain Barrier , Brain , Central Nervous System , Dendritic Cells , Encephalitis, Japanese , Interleukin-17 , Leukocytes , Lymphoid Tissue , Mice, Transgenic , Monocytes , Permeability , Phenotype , T-Lymphocytes , T-Lymphocytes, Regulatory , Th17 CellsABSTRACT
Objective:To explore the rule of CD8+CD25+FoxP3+ regulator T cells(Treg) in the pathogenesis of pre-eclampsia (PE) in peripheral blood.Methods:This study included 24 gestational-age-matched healthy pregnant women and 46 pregnant women diagnosed with mild PE (MPE,n=24) or severe PE (SPE,n=22) during the third trimester of gestation.An 3 ml sample of peripheral blood was drawn from each subject and anti-coagulated with heparin sodiurm The percentage of CD8 + CD25 + FoxP3 + Treg cells was detected by flow cytometry.The cytokines IL-6,IL-17A,IL-10,IL-1,IL-33 and TGF-β31 were detected using Luminex200.Peripheral blood mononuclear cells (PBMCs) were isolated frum healthy controls and treated with IL-33,the percentages of CD8+ CD25+ FoxP3 + Treg cell were measured by flow cytometric detection.Results:Compared to that of healthy pregnant controls [0.48(0.21-0.96)％],MPE patients [0.32(0.19-0.63)％] and SPE patients [0.13(0.02-0.41)％] had lower percentages of CD8+CD25+FoxP3+Treg cells (P＜0.05).Compared to HP controls,higher levels of IL-6 and IL-17A were found in MPE and SPE patients(P＜0.05) and even higher in SPE patients.The levels of IL-10,IL-1[β and IL-33 were similar in all three groups (P＞0.05).Compared to HP contruls,the levels of TGF-[β1 was significantly increased in SPE and MPE patients(P＜0.05),but no significant differences were found between these two groups (P ＞ 0.05).The percentage of CD8+ CD25+ FoxP3+ Treg cells showed a negative correlation with the serum concentrations of IL-17A (r =-0.338,P =0.338),and a positive correlation with the serum concentrations of IL-33 (r =0.548,P =0.548).After PBMCs were treated with IL-33 for five days,the percentages of CD8+CD25+FoxP3+ Treg were significantly higher than those of the contruls (P＜0.05).Conclusion:These findings suggested that the reduced CD8+ CD25 + FoxP3 + Treg cells may play a role in the pathogenesis of ore-eclamosia.
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Objective: To investigate the effect of Yupingfeng on hyaluronic acid (HA) and Foxp3+ Treg in patients with nasopharyngeal carcinoma. Methods: A total of 58 cases of nasopharyngeal carcinoma were divided into two groups, 30 cases in the treatment group, 28 cases in the control group. Patients in two groups were treated with synchronous radiotherapy and chemotherapy treatment, the treatment group was treated with the Yupingfeng granules through oral administration, 10 g/time, tid for 2 courses. The serum Foxp3+ Treg markers of each group were detected by flow cytometry assay before treatment and after treatment, and the level of HA in serum was detected by radio immunoassay. Results: After radiotherapy and chemotherapy, the contents of Foxp3+ Treg and HA were significantly decreased in two groups (P + Treg and HA (P + Treg and HA in nasopharyngeal carcinoma patients. Yupingfeng can also effectively reduce the side effect due to radiation and chemotherapy.
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OBJECTIVE@#To investigate the effect of Yupingfeng on hyaluronic acid (HA) and Foxp3(+) Treg in patients with nasopharyngeal carcinoma.@*METHODS@#A total of 58 cases of nasopharyngeal carcinoma were divided into two groups, 30 cases in the treatment group, 28 cases in the control group. Patients in two groups were treated with synchronous radiotherapy and chemotherapy treatment, the treatment group was treated with the Yupingfeng granules through oral administration, 10 g/time, tid for 2 courses. The serum Foxp3(+) Treg markers of each group were detected by flow cytometry assay before treatment and after treatment, and the level of HA in serum was detected by radio immunoassay.@*RESULTS@#After radiotherapy and chemotherapy, the contents of Foxp3(+) Treg and HA were significantly decreased in two groups (P < 0.05), and the decrease of treatment group was more significantly (P < 0.01). Correlation analysis showed positive correlation between Foxp3(+) Treg and HA (P < 0.05). After treatment, the incidence of side effects in two groups was significantly decreased. And there was significant difference between two groups (P < 0.05).@*CONCLUSIONS@#Combined chemotherapy and radiotherapy with Yupingfeng treatment can decrease the levels of Foxp3(+) Treg and HA in nasopharyngeal carcinoma patients. Yupingfeng can also effectively reduce the side effect due to radiation and chemotherapy.
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Objective:To investigate the effect of the adoptive transfer of CD4+CD25+Foxp3+regulatory T cells ( iTregs) induced by 5-aza-2′-deoxycytidine (5AzaD) on pregnant outcome of the abortion-prone mice.Methods:Sixty cases of female CBA/J × male DBA/2J abortion-prone matings were taken as study group,the CD4+T cells from spleen of twenty female CBA/J mice were separated by magnetic activated cell sorting (MACS),5AzaD was applied to the conversion of CD4+CD25-T cells to iTregs,the expression of Foxp3 in Tregs was characterized by flow cytometry analysis before and after epigenetic modification.The purified iTregs were injected into abortion-prone mice on day 1 or 4 of pregnancy,respectively,which were used as therapy groups,and then the embryo resorption rate was counted on day 14 of pregnancy.Results:After the treatment of 5AzaD,the percentage of iTregs in CD4+T cells was (41.50±8.03)%.The embryonic absorption rates of the two therapy groups were 10.47%(on day 1 of pregnancy) and 21.69%(on day 4 of pregnancy) ,respectively ( P<0.05 ) .Conclusion: Epigenetic modication of CD4+CD25-T cells may solve the problem of nTregs deficiency,particularly adoptive therapy of 5AzaD-induced iTregs at early stage of pregnancy can maintain normal pregnancy.
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Objective To explore the effect of Lactobacillussalivariuson the number of CD4+CD25+Foxp3+Treg cells and expression of transform?ing growth factorβ1(TGF?β1)in asthma Balb/c mice. Methods Thirty?two female Balb/c mice were randomly divided into four groups:the nor?mal control group,the asthma group,the Lactobacillus salivarius group,and the asthma combined Lactobacillussalivariusgroup. Acute asthma mod?el was established by the ovalbumin challenge method. After extraction of primary spleen cells,flow cytometry was used to test CD4+CD25+Foxp3+Treg/CD4+T ratio in spleen lymphocytes. The levels of IL?4,IFN?γand TGF?β1 in the spleen cell culture supernatant were measured by ELISA method. Results The level of Th2 cytokine(IL?4)in the spleen cell culture supernatant of the asthma group was significantly higher than that of the control group(P<0.05),however,the level of Th1 cytokine(IFN?γ)was significantly lower than that of the control group(P<0.05). The ex?pression level of Th2 cytokine(IL?4)in the Lactobacillussalivariusintervention group was significantly decreased compared with the asthma group, and the Th1 cytokine(IFN?γ)expression level was elevated compared with the asthma group(P<0.05). The level of TGF?β1 in the Lactobacillus salivarius intervention group was higher than in the asthma group(P<0.05). The proportion of CD4+CD25+Foxp3+Treg/CD4+T in spleen lympho?cytes in the asthma group was lower than that in the control group(P<0.05),and was higher in the Lactobacillus salivarius intervention group than in the asthma group(P<0.05). Conclusion CD4+CD25+Foxp3+Treg was associated with the pathogenesis of asthma. Lactobacillus salivarius could adjust Th1/Th2 imbalance and reduce asthma inflammation through up?regulation of CD4+CD25+Foxp3+Treg and TGF?β1 expression.
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Objective To investigate the regulatory effects of IFN-γon Treg cells from HIV/AIDS patients receiving highly active antiretroviral therapy (HAART) for one year.Methods Thirty HIV/A1DS patients whose CD4+T cells were below 350/μ1 were recruited for HAART therapy.Blood samples were collected at the time points of 0,24,48 weeks after HAART.PBMCs were isolated and randomly divided into two culture groups.One group was cultured directly in medium and another group was co-cultured with IFN-γ (40 pg/ml).The supernatants and cells were separated after 5 days of culture for analysis.The concentrations of IL-12 and CD4+CD25+Foxp3 Treg cells were measured by ELISA and flow cytometry,respectively.Results The levels of IL-12 in the supernatants from the culture without IFN-γ at time points of 0,24,48 weeks after HAART were lower than those from the co-cultured group [(37.02±12.76) vs (41.79± 15.02),t=2.336,P=0.03; (41.76±17.01) vs (47.2±14.26),t=2.702,P=0.014; (48.01± 11.84) vs (53.44± 11.30),t =3.14,P =0.003].The percentages of CD4+ CD25 + Foxp3 Treg cells in CD4+ T cells from the direct-cultured group were higher than those from the co-cultured group at the three time points [(10.41±1.10)％ vs (2.40±1.11)％,t=13.89,P=0.000; (8.33±2.03)％ vs (1.99± 0.86)％,t=12.93,P=0.000; (5.65±1.55)％ vs (1.32±0.73)％,t=10.61,P=0.000].Moreover,the results within the same group at the time points of 0,24,48 weeks upon HAART were also significantly different.Conclusion With the interference of HAART,IL-12 levels were increased,while CD4+CD25+ Foxp3 Treg cells were decreased in patients with HIV/AIDS.IFN-γ plays an important role in this process.
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Objective To observe the changes of number and proportion of CD4+CD25+FOXP3+ cells in peripheral blood of active rheumatoid arthritis patients (RA),and explore the function of recombinant human TNFR Ⅱ-Fc on the CD4+CD25-FOXP3+ Treg cells.Methods ① Forty severe RA patients were selected,who were divided into the combined treatment group (TNFR Ⅱ-Fc+MTX) and MTX only group according to the principle of randomized,double-blind,parallel and placebo-controlled study.All patients were treated for 12 weeks.Flow cytometry was used to analyze and compare the expression ratio of CD4+CD25+FOXP3 +Treg cells of RA patients' peripheral blood.Forty healthy controls were selected for parallel comparison.② VAS,DAS28,HAQ average of the two groups at different periods were compared.Matched t test was used to examine the quantity data between the groups.Results ① The proportion of CD4+CD25+FOXP3+ cells in the peripheral blood of active rheumatoid arthritis patients was significantly lower than healthy group [ (5.4±1.4)％ vs ( 7.5±1.5 )％,P＜0.01 ].The proportion of CD4+CD25+ FOXP3+ Treg cells in combined treatment group was significantly higher [(7.0+1.2)％ vs (5.2±1.6)％,P＜0.01 ] after TNFR Ⅱ-Fc and MTX combination therapy for 12 weeks.The increase rate of CD4+CD25+FOXP3+Treg cells in combined treatment group was evidently more remarkable than MTX only group [ (7.0 ± 1.2)％ vs (5.6 ±0.7 )％,P＜0.01].② After 12 weeks treatment,the arerage scores of VAS,DAS28 and HAQ of the combined treatment group were better than MTX only group,and the difference was statistically significant (P＜0.01).Conclusion This study has shown that the healing effect of TNFR Ⅱ -Fc combined with MTX is better than MTX only.TNFR Ⅱ -Fc can restore and improve the proportion of CD4+CD25+FOXP3+ Treg cells in active RA patients,which is likely to be an important mechanism for the treatment of RA patients.