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Objective:To investigate the effect of interleukin (IL)-7 receptor α (IL-7Rα) antibody on the immune inflammation and renal injury in MRL/lpr lupus mice.Methods:Fifteen 3-4-week-old female MRL/lpr lupus mice (specific pathogen free) weighing 15-16 g were bred to 14-week-old and randomly divided into three groups: IL-7Rα antibody intervention group, isotype antibody (positive control) group and normal saline (negative control) group. The mice in the threc groups were intraperitoneally injected with IL-7Rα antibody, isotype antibody and normal saline respectively, with 100 μg three times a week for 4 weeks. At the age of 18-week old, the mice were sacrificed. Twenty-four-hour urinary protein was detected by Coomassie brilliant blue method, serum creatinine was detected by peroxidase method, and the expression of autoantibody (anti-double strand DNA antibody) and inflammatory factors such as tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-21 was detected by enzyme-linked immunosorbent assay method. Renal pathology was detected by PAS and Sirius red staining, and CD3 and F4/80 in renal tissues were detected by immunohistochemistry method. Regulatory T cells, follicullar helper T cells (Tfh) and follicular regulatory T cells (Tfr) were detected by flow cytometry.Results:The 24-hour urinary protein, serum creatinine, serum anti-double strand DNA antibody and serum IFN-γ and IL-21 in the IL-7Rα antibody intervention group were significantly lower than those in the control groups (all P<0.01). However, there was no significant difference in serum TNF-α among the three groups ( F=0.39, P>0.05). The positive infiltrating cells of CD3 and F4/F80, and the ratio of type Ⅰ/Ⅲ collagen fibers ( F=41.11, P<0.01) of renal tissues in the IL-7Rα antibody intervention group were lower than those in the other two groups. Compared with the control groups, the ratio of regulatory T cells (CD4 +CD25 +Foxp3 +)/effector T cells (CD4 +CD25 +) in blood of IL-7Rα antibody intervention group increased ( F=21.64, P<0.01), while the ratio of Tfr (CD4 +CXCR5 +Foxp3 +)/Tfh (CD4 +CXCR5 +) in peripheral blood and spleen increased ( F=38.95, P<0.01; F=12.90, P<0.01). Conclusion:IL-7Rα antibody can reduce the production of autoantibodies such as anti-double strand DNA antibody and inflammatory factors by increasing the ratio of regulatory T cells and Tfr/Tfh, thus alleviating immune inflammation and renal damage in MRL/lpr lupus mice.
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Objective:To investigate the modulatory function of interleukin-7 (IL-7)/CD 127 signaling pathway on CD 8+T cells in patients with myasthenia gravis (MG). Methods:Fifty-seven treatment-naive MG patients who were hospitalized in Department of Neurology, Nanyang Central Hospital between 2017 and 2020 as well as 35 healthy controls were enrolled. Peripheral blood was collected, while plasma and peripheral blood mononuclear cells were isolated. Plasma IL-7 and soluble CD 127 (sCD 127) were measured by enzyme linked immunosorbent assay (ELISA). Membrane-bound CD 127 (mCD 127) percentage in CD 8+T cells was measured by flow cytometry. The differences of above indices between different gender, onset age, afflicting with thymoma, or different Osserman type and their correlation with Quantitative Myasthenia Gravis (QMG) score were analyzed. Purified CD 8+T cells from MG patients were stimulated with recombinant human IL-7 (5 μg/L). Changes of sCD 127 and mCD 127 level were analyzed. Levels of perforin, granzyme B, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) in the cultured supernatants were measured by ELISA. Immune checkpoint molecules mRNA in CD 8+T cells was semi-quantified by real-time fluorescence quantitative polymerase chain reaction. Results:Plasma IL-7 level was up-regulated in MG patients compared with controls [(293.4±74.7) pg/ml vs (233.8±70.8) pg/ml, t=3.78, P<0.001], while sCD 127 level was down-regulated in MG patients compared with controls [(102.7±13.7) pg/ml vs (131.2±20.9) pg/ml, t=7.91, P<0.001]. Peripheral CD 8+T cells percentage was up-regulated in MG patients compared with controls (35.4%±7.1% vs 30.2%±7.5%, t=3.31, P=0.001), and mCD 127+CD 8+T cell percentage was also elevated (45.5%±7.7% vs 34.7%±11.5%, t=5.44, P<0.001). There were no significant differences of above indices between different gender, onset age, afflicting with thymoma, or different Osserman type. There was no significant correlation between above indices and QMG score. There were no significant differences of sCD 127 in cultured supernatants, mCD 127+CD 8+T cell percentage, or immune checkpoint molecules mRNA expression between CD 8+T cells from MG patients with and without IL-7 stimulation. IL-7 stimulation promoted the secretion of perforin [(208.1±67.2) pg/ml vs (168.8±46.2) pg/ml, t=2.16, P=0.038], granzyme B [(941.8±273.9) pg/ml vs (782.4±137.2) pg/ml, t=2.33, P=0.025], and IFN-γ [(19.1±5.2) pg/ml vs (15.3±4.5) pg/ml, t=2.47, P=0.018] by CD 8+T cells. However, there was no remarkable difference of TNF-α production between CD 8+T cells with and without IL-7 stimulation. Conclusion:Elevated IL-7-mediated signaling pathway enhanced the secretion of cytotoxic molecules and cytokines by CD 8+T cells, leading to increased activity of CD 8+T cells in MG patients.
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Objective To explore the effect of anti-retroviral therapy on interleukin(IL)-7/IL-7R in human immunodeficiency virus(HIV) infected patients in China.Methods Cases were divided into 2 groups:HIV-infected group (35 cases),and control group (30 cases).IL-7 in serum,IL-7R(CD127) expression in CD4 +T cells,and CD4 +T cells count were detected and compared between two groups before and after treatment for 1 year.Results IL-7 level in the serum of HIV infected group before treatment [(8.98 ±3.77) pg/ml] was significantly higher than that in control group [(3.84 ±0.86) pg/ml] (P ＜0.05).The counts of CD4+T cells [(202.65 ± 121.54)/μl],CD4 + CD127 + T cells [(60.25 ± 11.75) ％],and CD8 + CD127 + T cells [(46.27 ± 12.10)％] in HIV-infected group were significantly lower than those in control group [(766.99 ± 103.21)/L,(76.89 ± 20.01) ％,(81.27 ± 12.35)％] (P ＜0.05).After anti-retroviral therapy (ART),IL-7 level in the serum of HIV-infected group[(5.55 ± 1.35) pg/ml]was decreased,and CD4+T cells [(450.58 ± 15)/μl],CD4 + CD127 +T cells [(69.82 ± 15.24)％],and [CD8 + CD127 + T(59.23± 14.73) ％] cells was increased in HIV-infected group,with a significant difference between two groups (P ＜0.05).Conclusions ART could improve the IL-7 level in the serum and IL-7R(CD127)expression in CD4 +T cells of HIV-infected patients.However,they still cannot become normal level.
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Objective To compare the effects between the impact of IL-7Rα,bacterial flagellin alone to the acute graft-versus-host disease and the combination of both,and to explore its possible mechanism.Methods The BALB/C (H-2d) female mice as recipients were divided into alone irradiation transplantation group (group A),IL-7Rα intervention group (group B),bacterial flagellin intervention group (group C),IL-7Rα combined with bacterial flagellin intervention group (group D),and 6 mice in each group.All mice were accepted 9 Gy 60Co total body irradiation,and 1×107 bone marrow cells and 2× 107 spleen cells of donors C57BL/6 (H-2b) mice were infused via the tail vein to recipient mice.The symptoms,signs,survival time and hematopoietic function recovery of the recipient mice were observed.Results Mice survival of group A was (22.5±2.30) d,30 d survival rate was 50.0 ％ (3/6),and aGVHD performances inculding the fatigue,anorexia,hair removal,arched,and so on appeared obviously.Survival of group B was (25.83±3.49) d,30 d survival rate was 33.3 ％ (2/6),aGVHD performances compared with group A was lighter.Survival of group C was (26.33±3.52) d,30 d survival rate was 33.3 ％ (2/6),also appeared aGVHD performance,which degree was same to the group B.survival of group D was (30.17±2.86) d,30 d survival rate was 66.7 ％ (4/6),aGVHD performances compared to the other three groups was lightest.The white blood cell count of four groups were reduced to minimum at +7 d,then the three intervention groups gradually recovered.The WBC recovery at 14,21,28,30 day after the transplant of group A compared with slowly was the intervention groups (P ＞ 0.05),WBC recovery of B was roughly equal to group C (P ＞ 0.05),while the WBC recovery of group D was faster than group B or C (P ＜ 0.05).At 2nd week after transplantation,CD3+ T cells was significantly decreased in 4 groups,and at 3rd week began gradually rised.Compared with group A,the proportion of CD3+ cells of other three groups were increased significantly,there was no statistical signifiance of CD3+ cell proportion between group B and group C at 2nd,3rd,4th week after transplantation (P ＞ 0.05),while the CD3+ T cell recovery in group D was faster than group B or C (P ＜ 0.05).Conclusions The stable aGVHD mouse model was established.Exogenous IL-7Rα and bacterial flagellin may reduce the incidence of aGVHD.There was no significant difference for aGVHD when they was used alone,but when combination of them,aGVHD is slighter and the hematopoietic recorery is faster.
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Objective To analyze CD127 expression on the memory CD8+ lymphocytes from hepatitis B e antigen (HBeAg) positive chronic hepatitis B (CHB) patients treated with peginterferon α-2a (Pegasys). Methods Thirty HBeAg positive CHB patients were treated with peginterferon α-2a 180 μg once a week for 48 weeks and followed up for 24 weeks. The memory CD8+ lymphocytes were characterized by expressing CD45RA and CD27 markers. CD127 expression on cell surface was measured by four-colour flow cytometry. The difference of mean values between groups was evaluated by Mann-Whitney test. Results The CD127 expression on CD8+ T lymphocytes was significantly lower in HBeAg positive CHB patients compared to healthy controls (Z=2.889, P＜0.05), which was negatively correlated with serum hepatitis B virus (HBV) DNA level and HBeAg titers. The CD127 expression increased along with the decrease of HBV DNA and HBeAg after 24-week, 48-week and 72-week treatment in patients showing good response to peginterferon α-2a, while CD127 expression didn't change markedly in non responders (Z24w = 1.954, Z48w = 2.789, Z72w = 2. 989; all P＜0. 05). Conclusion CD127 expression on memory CD8+ lymphocytes increases along with effective anti-HBV treatment in CHB patients, which can be used as a marker for evaluating the effectiveness of anti-viral treatment.