CD4 T lymphocyte subsets displa heterogeneous susceptibility to apoptosis induced by serum from patients with systemic lupus erythematosus

Abstract Background Serum from systemic lupus erythematosus (SLE) patients has been shown to induce T-lymphocyte (TL) apoptosis. Given that different cells of the immune system display different sensitivity to apoptosis, we set to evaluate the in vitro effect of SLE serum on regulatory T-cells (Treg), Th17, Th1 and Th2 from SLE patients and healthy controls. Methods Peripheral blood mononuclear cells from SLE patients or normal controls were exposed to a pool of sera from SLE patients or normal controls. Annexin V was used to label cells in apoptosis or necrosis. Annexin V-labeled Treg, Th17, Th1 and Th2 cells were determined using flow cytometry. Results Total CD3 + and CD4+cells from SLE patients showed higher frequency of spontaneous apoptosis/necrosis, whereas Th1 cells from SLE patients presented reduced spontaneous apoptosis/necrosis rate as compared with cells from controls. Incubation with SLE serum induced increased frequency of apoptotic/necrotic CD3 +, CD4 + and Th2 cells from normal controls or from SLE patients as compared with cultures incubated with normal human serum (NHS) or without human serum at all. Incubation with SLE serum did not increase the apoptosis/necrosis rate in Th1 or Th17 cells. Treg cells from SLE patients were more prone to apoptosis/necrosis induced by SLE serum than Treg cells from normal individuals. Th1, Th2, and Th17 cells presented increased apoptosis rates in cultures without human serum. Conclusion Our findings indicate that the serum of patients with active SLE stimulates apoptosis of CD4+T cells in general and exhibit differentiated effects on CD4+T-cell subsets.

Immunomodulatory mechanism of interleukin-7 receptor α antibody attenuating renal injury in MRL/lpr lupus mice / 中华肾脏病杂志

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.

Changes of Th17/Treg and serum cytokines in peripheral blood of patients with acute exacerbation of chronic obstructive pulmonary disease and secondary pulmonary fungal infection / 中国综合临床

Objective:To explore the changes of peripheral blood Th17/Treg and serum cytokines in AECOPD patients with secondary pulmonary fungal infection.Methods:Selected the clinical data of 27 AECOPD patients who were admitted between January 2018 to March 2020 in the Department of Respiratory Medicine, Hai'an People's Hospital Affiliated to Nantong University with fungal infection (fungal infection group), and 58 AECOPD patients without fungal infection (non-fungal infection group) who received treatment in the hospital during the same period. Compared the general clinical data, peripheral blood Th17 and Treg cell levels, Th17/Treg ratio, interleukin-17 (IL-17), interleukin-23 (IL-23), interferon-γ (interferon-γ, IFN-γ), and transforming growth factor-β (TGF-β) levels. Meanwhile, compared the levels of Th17 and Treg cells in peripheral blood, the ratio of Th17/Treg and serum cytokines in patients with different infection severity in fungal infection group. The measurement data with normal distribution were compared by independent samplet t-test between the two groups, one-way ANOVA between multiple groups, LSD-t test for pairwise comparision, and χ 2 test for counting data. Results:In the 27 AECOPD patients with fungal infection group, the pathogen distribution was 65.52% (19/27) of candida albicans, 10.34% (3/27) of candida tropicalis,10.34% (3/27) of candida albicans, and 6.90% (2/27) of Aspergillus. The level of Th17 [(16.18±3.15) % and (12.34±2.64) %, t=5.87, P<0.001)], the ratio of Th17/Treg [(4.70±0.85) and (2.41±0.51), t=22.87, P<0.001] in Patients with fungal infection group were higher than those in the non-fungal group. The level of Treg [(3.42±0.42) % and (5.13±0.51) %, t=20.77, P<0.001] in Patients with fungal infection group was lower than those in the non-fungal group. The levels of IL-17 [(85.67±21.51) μg/L and (53.64±14.36) μg/L, t= 8.12, P<0.001], and IL-23 [(61.38±16.58) μg/L and (38.29±12.60) μg/L, t=7.10, P<0.001] in Patients with fungal infection group were higher than those in non-fungal infection group, but the levels of IFN-γ ((47.75±17.72) μg/L and (62.37±19.06) μg/L, t=3.37, P=0.001) and TGF-β ((110.34±26.03) μg/L and (131.40±35.03) μg/L, t=2.87, P=0.007) were lower than those in non-fungal infection group, and the differences were statistically significant. There were statistically significant differences in the ratio of Th17/Treg, and the levels of Th17, Treg cells and cytokine among patients with different infection severity in the fungal infection group. With the increase of infection severity, the levels of Th17 ((13.06±1.98)%, (15.94±2.11)%, (17.75±2.20)%, F=10.19, P<0.001), the ratios of Th17/Treg ((5.01±0.60), (5.66±0.69), (6.52±0.65), F=10.77, P<0.001), the levels of IL-17 ((63.39±11.64) μg/L,(78.66±12.82) μg/L, F=9.01, P=0.001), and IL-23 ((42.52±13.11) μg/L, (59.97±15.25) μg/L, (69.75±14.30) μg/L, F=7.41, P=0.003) were increase, the levels of Treg ((4.33±0.39)%, (3.32±0.42)%, (2.50±0.35)%, F=44.42, P<0.001), IFN-γ ((57.78±10.52) μg/L, (48.82±10.39) μg/L, (38.90±10.56) μg/L, F=6.50, P=0.006), TGF-β ((126.62±18.94) μg/L, (115.34±13.66) μg/L, (102.52±17.73) μg/L, F=4.25, P=0.026) were significantly decreased. Conclusion:The imbalance of Th17/Treg ratio and related serum cytokines play an important role in the process of lung fungal infection in AECOPD patients, and their imbalance is related to the severity of fungal infection. Therefore, the levels of Th17/Treg and serum cytokines should be closely monitored in AECOPD patients.

Changes of Th17/Treg proportion and the expressions of related cytokines in peripheral blood of patients with Hashimoto's thyroiditis / 中华地方病学杂志

Objective:To observe the ratio of helper T cells 17 (Th17)/regulatory T cells (Treg) in peripheral blood of patients with Hashimoto's thyroiditis (HT) and the expression changes of related cytokines in serum, and to explore their role in the occurrence and development of HT.Methods:Using the case-control study method, 35 HT patients examined in the General Hospital of Heilongjiang Beidahuang Group from February to November 2019 were selected as HT group, and 39 healthy people in the same period were selected as control group. Early morning fasting venous blood samples of the two groups were collected to test the levels of thyroid stimulating hormone (TSH), free thyroxine (FT 4), thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb). The expressions of serum interleukin (IL)-6, IL-17 and transforming growth factor β (TGF-β) were tested by enzyme-linked immunosorbent assay (ELISA); the number of Th17, Treg in peripheral blood were determined by flow cytometry. Results:The levels of TPOAb, TgAb and TSH in HT group [130.60 (43.37, 714.40), 368.10 (136.90, 1 103.00) U/ml, 9.05 (6.62, 15.23) μU/ml] were significantly higher than those in control group [2.66 (1.52, 4.69), 12.63 (11.43, 14.60) U/ml, 1.87 (1.36, 2.23) μU/ml, U = 6.87, 6.62, 4.85, P < 0.001], and the FT 4 level [0.76 (0.63, 1.04) ng/dl] was lower than that in control group [1.14 (1.02, 1.26) ng/dl, U = 7.39, P < 0.001]. The expressions of IL-6, IL-17 and TGF-β in HT group were higher than those in control group ( t = 2.41, 9.04, 2.44, P < 0.05). The number of Th17 and the ratio of Th17/Treg in HT group were higher than those in control group ( t = 4.20, 3.50, P < 0.05), and the number of Treg was lower than that in control group ( t = 4.45, P = 0.001). Conclusion:In HT patients, Th17 are increased, Treg decreased, Th17/Treg ratio increased, and the expressions of IL-6, IL-17 and TGF-β are increased.

Advances on follicular helper T cells in patients with hepatitis B virus infection / 中华临床感染病杂志

Hepatitis B virus (HBV) infection is a major global public health problem. Persistent HBV infection is prone to develop chronic hepatitis B (CHB), and CHB is closely related to the development of liver fibrosis and hepatocellular carcinoma. High-affinity specific anti-HBs are essential for the control of HBV infection, while the antibody production is closely related to follicular helper T (Tfh) cells. Tfh cells can help B cells differentiate into plasma cells to produce specific antibodies to control virus infection. This article reviews the latest research progress of Tfh cells in HBV infection to provide information of new strategies for the prevention and treatment of HBV.

Intervention effects and mechanism of Compound yu ’e nasal drops on allergic rhinitis in rats / 中国药房

OBJECTIVE To study the intervention effects and mechanism of Compound yu ’e nasal drops on ovalbumin induced allergic rhinitis in rats . METHODS The allergic rhinitis model of rat was induced with ovalbumin . Model rats were randomly divided into model group ，triamcinolone acetonide group （positive control ，0.026 mg/kg），Compound yu ’e nasal drops high-dose，medium-dose and low -dose groups （134.4、67.2、33.6 mg/kg），12 rats in each group . Another blank control group was set. Except for blank control group ，the corresponding drugs were given by nasal drip twice a day for 14 days. One hour after last administration，the nasal symptom scores of rats were recorded ；the levels of serum immunoglobulin E （IgE），interleukin-2（IL- 2），IL-13 and tumor necrosis factor -α（TNF-α）were measured by enzyme -linked immunosorbent assay . The changes of nasal mucosa in rat were observed by HE staining . The expressions of TNF -α，IL-2 and IL -13 in nasal mucosa were detected by Western blot. RESULTS Compared with blank control group ，nasal symptom score and the levels of serum IgE ，IL-2，IL-13，TNF-α in model group were increased significantly （P＜0.01）；obvious pathological injury was found in nasal mucosa ，and the expressions of TNF -α，IL-2 and IL -13 protein were increased significantly （P＜0.01）. Compared with model group ，Compound yu ’e nasal drops significantly reduced the nasal symptom score ，the levels of serum IgE ，IL-2，IL-13，TNF-α to different extents ，improved pathological injury of nasal mucosa and significantly inhibited the expressions of TNF -α，IL-2 and IL -13 protein（P＜0.05 or P＜ 0.01）. CONCLUSIONS Compound yu ’e nasal drops play significant effects against allergic rhinitis in rats by regulating the balance of t ype 1 helper T cells/type 2 helper T cells ，balancing and inhibiting the secretion of inflammatory cytokines .

Research progress on OX40/OX40L axis mediated T cell immunity in the course of oral lichen planus / 口腔疾病防治

@#Oral lichen planus (OLP) is a common chronic disease of the oral mucosa with unclear pathogenesis. Local infiltration of T cells plays a key role in the pathological process of OLP. Increased evidence supports the notion that the imbalance of helper T cells (Th) 1/Th2 and Th17/regulatory T cells (Treg) and their related cytokines is closely related to the pathogenesis and progression of OLP. In recent years, studies have shown that OX40 (CD134) and its ligand OX40L (CD252) play an important role in the process of the T-cell immune response. They participate in the balance regulation of Th1/Th2 and Th17/Treg, mediate the imbalance of pro-inflammatory and anti-inflammatory, and affect the occurrence and development of a variety of autoimmune diseases. However, there is no direct evidence that the OX40/OX40L axis mediates the imbalance of T-cell subsets in the pathogenesis of OLP. Therefore, large sample clinical as well as in vitro and in vivo experimental studies on the mechanism by which the OX40/OX40L axis regulates the balance of T-cell subsets in OLP are still needed in the future.

Follicular Helper T Cells， Psoriasis， and Chinese Medicine Intervention： A Review / 中国实验方剂学杂志

Follicular helper T （Tfh） cells are a newly discovered subset of CD4+ T cells. As reported， abnormalities in their development， differentiation， and function are closely related to the occurrence of autoimmune diseases. Psoriasis is an autoimmune skin disease and it is intractable with a prolonged course. At present， it is generally believed that immune imbalance mediated by T cells is the core mechanism of the pathogenesis of psoriasis. In the context of this mechanism， Tfh cells are associated with psoriasis， and their cellular level and abnormal expression of related candidates can promote the occurrence of psoriasis. In terms of treatment， Chinese medicine， by virtue of the characteristics of wide application and low price， serves as a good complementary and alternative treatment option for psoriasis. As confirmed by previous findings， some active ingredients or preparations of Chinese medicine used in the treatment of psoriasis can also intervene in and regulate the immune response mediated by Tfh cells and the related candidates. Based on the research reports and experimental data， the present study reviewed the research progress from the differentiation of Tfh cells， the relationship between Tfh cells and psoriasis， and the intervention and regulation of Tfh cells and related molecules by Chinese medicine， which is expected to provide certain theoretical support and references for the determination of new strategies for psoriasis treatment and research in related fields.

First report of expansion of CD4+/CD28 null T-helper lymphocytes in adult patients with idiopathic autoimmune hemolytic anemia

ABSTRACT CD28 null T helper (Th) cells are rare in healthy individuals, but they are increased in various inflammatory and immune-mediated diseases. In this study, we determined the size of the CD4+/CD28 null T lymphocyte compartment in the peripheral blood of 40 autoimmune hemolytic anemia (AIHA) patients (idiopathic and secondary) and 20 healthy control subjects, using tri-color flow cytometry. The frequency and absolute count of CD4+/CD28 null T helper (Th) cells was significantly higher in idiopathic AIHA patients, compared to healthy controls (p = 0.001 and 0.001, respectively) and to patients with secondary AIHA (p = 0.04 and 0.01, respectively). The percentage of CD4+/CD28 null Th cells was also negatively correlated to the hemoglobin (Hb) level (p = 0.03). These findings demonstrate, for the first time, the expansion of this phenotypically-defined population of T lymphocytes in patients with idiopathic AIHA and indicate that it likely plays an etiological role in the development of this disease. However, establishing the use of this marker for diagnosis or monitoring treatment of such patients needs further studies.

The significance of follicular helper T cells and galactose-deficient IgA 1 in the pathogenesis of Henoch-Sch?nlein purpura in children / 国际儿科学杂志

Objective:To investigate the role of follicular helper T(Tfh) cells and galactose deficiency IgA 1(Gd-IgA 1) in the children that were suffering from Henoch-Sch?nlein purpura(HSP) and Henoch-Sch?nlein purpura nephritis(HSPN)and the correlation between them. Methods:According to the presence or absence of renal injury, 62 children with HSP were divided into HSP group with 32 children and HSPN group with 30 children.Twenty children who underwent physical examination at outpatients were known as the healthy control group.Flow cytometry was used to measure the proportion of Tfh(CD4 + CXCR5 + PD-1 + ) in peripheral blood.Immunoturbidimetry and ELISA were used to measure the serum levels of IgA 1 and Gd-IgA 1 respectively. Results:(1) The proportion of Tfh cells in peripheral blood and the serum levels of Gd-IgA 1 in both HSP group and HSPN group had significantly increased than those in healthy control group( P<0.01). Compared result of the HSPN group with HSP group, the proportion of Tfh cells in peripheral blood and the serum levels of Gd-IgA 1 in HSPN group were higher than that in HSP group( P<0.05). (2) In the HSPN group, the proportion of peripheral blood Tfh cells and the serum levels of Gd-IgA 1 in group of renal pathology ≥ grade Ⅲ and heavy proteinuria were significantly elevated compared with group of renal pathology < grade Ⅲ and non-heavy proteinuria(<0.01). (3) In the healthy control group, the serum levels of Gd-IgA 1 was positively correlated with the proportion of Tfh cells in peripheral blood and the serum levels of Gd-IgA 1( P<0.05). Conversely, a non-positive correlation was shown in HSP and HSPN groups( P>0.05). Conclusion:The excessive activation of Tfh cells and the serum levels of Gd-IgA 1 may be one of the pathogenesis of HSP/HSPN, the degree of increment of the two factors may be related to the activity and severity of the disease.The mechanism of Tfh cells potentially leading to an increase of Gd-IgA 1 production requires further study.

Therapeutic effect of gene silencing peptidyl arginine deaminase 4 on pulmonary interstitial lesions induced by collagen-induced arthritis mice / 北京大学学报(医学版)

OBJECTIVE@#To investigate the therapeutic effect of gene silencing peptidyl arginine deaminase 4 (PAD4) on pulmonary interstitial lesions induced by collagen-induced arthritis (CIA) mice, and possible mechanisms.@*METHODS@#A CIA mouse model was established in DBA/1 mice, followed by a tail vein injection of the virus solution prepared by the PAD4-siRNA expression vector once a week for 8 times. The mice were sacrificed at the end of the experiment. The expression of PAD4 mRNA in lungs was detected by real-time quantitative PCR (qRT-PCR). The expression of PAD4 protein was detected by tissue immunohistochemistry. Cell culture was performed by spleen tissue. Flow cytometry changes in the ratio of Tfh cells to Tfr cells were examined; lung staining was performed in the lungs to observe changes in lung pathology.@*RESULTS@#(1) Compared with the blank group, the expression of PAD4 mRNA in the lung tissue of the model group increased, the difference was statistically significant (P < 0.05). PAD4 mRNA in the lung tissue of the CIA mice after PAD4-siRNA treatment. The expression level was significantly lower than that of the model group and the negative control group, and the difference was statistically significant (P < 0.05). (2) Red fluorescence was less in the lung tissue of the blank group, while more red fluorescence was observed in the inflammatory cell infiltration area and trachea around the lung tissue of the model group and the negative control group, and the red fluorescence of the three groups after PAD4-siRNA treatment was significantly reduced; (3) Compared with the blank group, the proportion of Tfh cells in the model group increased, the difference was statistically significant (P < 0.05), the proportion of Tfh cells in spleen cells of the CIA mice after PAD4-siRNA treatment was significantly lower than that of the model group and the negative control group, the difference was statistically significant (P < 0.05); compared with the blank group, in the mouse spleen cells in the model group the proportion of Tfr cells was slightly decreased, but the difference was not statistically signifi-cant. The proportion of Tfr cells in the spleen cells of the mice increased after PAD4-siRNA treatment, but the difference was statistically significant only in the PAD4-siRNA2 group compared with the model group and the negative control group (P < 0.05); (4) The proportion of Tfh/Tfr in the spleen cells of the model group was increased, compared with the blank group, the difference was statistically significant (P < 0.05); the ratio of Tfh/Tfr in the three groups after PAD4-siRNA treatment all decreased, the difference was statistically significant (P < 0.05); (5) Compared with the blank group, the alveolar wall of the lung tissue of the model group was thickened, the inflammatory cell infiltration was increased, and the lung tissue destruction and inflammatory infiltration of the CIA mice were decreased after PAD4-siRNA treatment. The degree of reduction was reduced.@*CONCLUSION@#Gene silencing of PAD4 can reduce the proportion of Tfh cells, increase the proportion of Tfr cells, reverse the proportion of Tfh/Tfr, and reduce the degree of interstitial lesions and inflammatory infiltration of lung tissue.

Observation of Effect of Qianjin Weijingtang on Differentiation of Th17/Treg and Expressions of Related Cytokines in Lung Tissues of Model Rats Exposed to Cigarette Smoke / 中国实验方剂学杂志

Objective:The present study was to observe the effect of Qianjin Weijingtang on the differentiation of helper T cells 17 （Th17）/T regulatory cell （Treg） and the expressions of related cytokines in the lung tissues of the model rats exposed to cigarette smoke. Method:Totolly 60 male rats were randomly assigned into six groups （control group， model group， acetylcysteine group and Qianjin Weijingtang high， moddle and low dose groups）， with 10 rats in each group. After 30 day's modeling and 30 day's intervention， rats were killed peacefully with their tissues collected. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression of retinoic acid associated orphan receptor （ROR-γt） and forkhead/pterygoid helix transcription factor 3 （Foxp3） mRNA， enzyme-linked immunosorbent assay （ELISA） was used to check the concentration of interleukin-17（IL-17）， IL-6， IL-10 and transforming growth factor-β1（TGF-β1）. Htoxylin eosin （HE） staining was used to observe the pathological changes of lung tissues， while flow cytometry was used to detect Treg（Foxp3+CD25+）， Th17（CD4+IL-17+） and Treg/Th17 ratio. Result:As compared with the control group， the ROR-γt mRNA expression in model group was higher（P<0.01）. As compared with the model group， various doses of Qianjin Weijingtang down-regulated the expression（P<0.05，P<0.01）. As compared with the control group， the expression of the Foxp3 mRNA was down-regulated in model group（P<0.05）， but was up-regulated in Qianjin Weijingtang middle and low dose groups（P<0.05）. Compared with the control group， the concentration of IL-17 and IL-6 in the model group were significantly increased， while the concentration of IL-10 and TGF-β1 in the model group were significantly decreased（P<0.05，P<0.01）. As compared with the model group， intervention with various doses of Qianjin Weijingtang could help to decrease the concentration of IL-17 and IL-6 in lung tissues， and increase the concentration of IL-10（P<0.05）， which were consistent with those of Real-time PCR results. Flow cytometry examination showed that the Th17（CD4+IL-17+） proportion decreased and Treg/Th17 ratio increased after Qianjin Weijing Tang middle and low dose intervention（P<0.05，P<0.01）. Conclusion:Qianjin Weijingtang could regulate Treg/Th17 ratio and help to achieve Th17/Treg balance.

Effect of scraping therapy on Th1/Th2 balance in rats with lumbar disc herniation / 针灸推拿医学（英文版）

Objective: To observe the effect of scraping therapy (ST) on the immune balance of serum helper T (Th) 1/Th2 cells in autologous nucleus pulposus transplantation non-compressive lumbar disc herniation (LDH) model rats, and explore the immune mechanism of ST in reducing LDH pain. Methods: Seventy-two male Sprague-Dawley (SD) rats were randomly divided into a model group, a sham operation group and a ST group, with 24 rats in each group. Autologous nucleus pulposus transplantation non-compressive LDH models were established in the model group and ST group, while the rats in the sham operation group underwent sham operation without model establishment. On the fifth day after the model was successfully prepared, rats in the ST group received ST, once every other day, 3 times as a course for a total of 3 courses. Six rats in each group were randomly selected to observe their pain thresholds, and peripheral blood of the rats was collected before the first scraping treatment and at the end of the first, second, and third courses of treatment. The serum was isolated and enzyme-linked immunosorbent assay (ELISA) was applied for the detection of rat serum interferon (IFN)-γ, interleukin (IL)-4 and IL-10. Results: Compared with the model group, the pain threshold in LDH rats in the ST group increased (P<0.05), the serum IFN-γ level was significantly reduced (P<0.05), but the levels of IL-4 and IL-10 did not change significantly (both P>0.05). At the end of the second and third courses of treatment, the IFN-γ/IL-4 ratio was negatively correlated with the pain threshold in the rats, and the IFN-γ/IL-4 ratio was significantly reduced in the ST group (P<0.01). Conclusion: ST can help suppress the Th1 immunity in LDH rats triggered by the autologous nucleus pulposus, restore the immune balance of Th1/Th2, and reduce the pain of LDH.

Therapeutic effects of baicalin on lupus nephritis in a lupus-prone mouse model and its regulatory role in follicular helper T cell differentiation / 中华皮肤科杂志

Objective To evaluate the therapeutic effect of baicalin on lupus nephritis in a lupusprone mouse model,and to investigate its regulatory role in the differentiation of follicular helper T (Tfh)cells.Methods Eight 12-week-old female MRL/lpr lupus-prone mice were randomly and equally divided into two groups by a random number table i.e.,baicalin group and control group intraperitoneally injected with 200 mg/kg baicalin sodium and chloride physiological solution,respectively,once every day for 4 weeks.After the end of treatment,Coomassie brilliant blue staining was performed to detect the level of 24-hour urine protein.Then,the mice were sacrificed,and the spleens were resected and weighed.Mononuclear cells were isolated from these spleens,and flow cytometry was conducted to determine the proportion of Tfh cells.Additionally,the kidneys were resected and subjected to hematoxylin and eosin (HE) staining for the evaluation of kidney impairment.Moreover,some other mononuclear cells were isolated from the spleens of the lupus-prone mice in the control group,and magnetic activated cell sorting (MACS) was performed to isolate naive CD4+ T cells,which were divided into 3 groups:blank control group receiving no treatment,induction group treated with 10 μg/L anti-interleukin (IL)-21 and anti-IL-6 antibodies and 3 μg/L anti-CD3 and anti-CD28 antibodies for 5 days,and intervention group additionally treated with 40 μmol/L baicalin for 5 days besides the above treatment.Then,50 μg/L phorbol ester,750 μg/L ionomycin and 20 mg/L brefeldin A were used to stimulate some cultured naive CD4+ T cells in the above groups.Flow cytometry was conducted to determine the proportion of CD4+CXCR5+PD-1 + cells and CD4+IL-21+ cells.Statistical analysis was carried out with SPSS20.0 software by using one-way analysis of variance (ANOVA) and student t test for the comparison of quantitative data between groups.Results The baicalin treatment could effectively improve the kidney impairment in the lupus-prone mice.Compared with the control group,the baicalin group showed significantly decreased 24-hour urine protein level ([1 416 ± 171] vs.[2 623 ± 278] μg/24 h,P =0.022),and significantly decreased proportion of Tfh cells in the spleen (12.6％ ± 2.3％ vs.40.2％ + 1.1％,P =0.005).In vitro baicalin could further inhibit the differentiation of Tfh cells.Compared with the induction group,the intervention group showed significantly decreased proportion of CD4+CXCR5+PD-1+ Tfh cells (13.3％ ± 0.8％ vs.17.6％ ± 0.9％,P =0.04) and CD4+IL-21+ cells (1.0％ ± 0.4％ vs.2.7％ ± 0.2％,P ＜ 0.01).Conclusion Baicalin can effectively ameliorate lupus nephritis,which may be associated with the inhibition of Tfh cell differentiation.

Activated Leukocyte Cell Adhesion Molecule Modulates Th2 Immune Response in Atopic Dermatitis

PURPOSE: Activated leukocyte cell adhesion molecule (ALCAM), a member of the immunoglobulin superfamily, is highly expressed on dendritic cells. ALCAM and its receptor CD6 are co-stimulatory molecules in the immunological synapse; their interaction is required for T cell activation. While atopic dermatitis (AD) is recognized as a T helper 2 (Th2)-mediated allergic disease, the role of ALCAM in its pathogenesis is unclear. METHODS: ALCAM levels were measured in the serum of AD patients and AD-induced murine model by ovalbumin treatment. We next investigated transepidermal water loss, clinical score, Th2-immune responses, skin barrier gene expression and T-cell activation using wild-type (WT) and ALCAM deficiency mice. An oxazolone-induced AD-like model was also established and analyzed using WT- and ALCAM-deficient mice. RESULTS: We found that serum ALCAM levels were elevated in pediatric AD patients as well as WT AD mice, whereas Th2-type cytokine production and AD symptoms were suppressed in ALCAM-deficient mice. In addition, CD4+ effector T-cell counts in murine skin and skin-draining lymph nodes were lower in ALCAM-deficient mice than in their WT counterparts. ALCAM deficiency was also linked to higher expression of skin barrier genes and number of lamellar bodies. CONCLUSIONS: These findings indicate that ALCAM may contribute to AD pathogenesis by meditating a Th2-dominant immune response and disrupting the barrier function of the skin.

Advances in Research on Intervention of Th17/Treg Balance in Ulcerative Colitis by Traditional Chinese Medicine / 中国实验方剂学杂志

Ulcerative colitis (UC) is a common chronic non-specific intestinal inflammation, which has a long course and is difficult to cure, and the incidence is increasing year by year. Helper T cells 17 (Th17) are one of immune-promoting cells, while regulatory sputum cells (Treg) are immunosuppressive cells, and Th17 cells and regulatory T cells together maintain the balance of the body's immune microenvironment. During progression of UC, the population of sputum helper cells 17 (Th17 cells) that cause inflammation generally increases, while the number of sputum regulatory T cells that inhibit Th17 cell activity decreases. Among them, Th17 cells mediate immune response, regulatory T cell-mediated immunosuppression, and the balance between the two plays a key role in the inflammation and immune process of ulcerative colitis. Although Western medicine treatment of UC has certain effects, however, The frequency and severity of side effects, inconvenient dose adjustments, and partial price excessions limit their clinical application.As a traditional medicine in China, traditional Chinese medicine(TCM) has multi-target, multi-link and multi-channel treatment characteristics, and has unique advantages and broad prospects in anti-ulcerative colitis. In recent years, the TCM field has taken Th17/Treg balance as the entry point, and carried out a large number of clinical and experimental studies on the intervention of TCM in Th17/Treg balance in UC, and achieved certain results. Clinical and experimental evidence clearly indicates intervention in Th17/Treg. It is an important mechanism of action of TCM in the treatment of ulcerative colitis. This paper mainly summarizes and analyzes the intervention effects of TCM monomer, component or active ingredient and TCM compound on Th17/Treg balance in UC, which is helpful for people to understand Chinese medicine intervention in UC more accurately and comprehensively. The mechanism of action of Th17/Treg balance provides a reference for the clinical design of a treatment plan for anti-ulcerative colitis.

Enhancement of T Follicular Helper Cell-Mediated Humoral Immunity Reponses During Development of Experimental Autoimmune Myasthenia Gravis / 神经科学通报·英文版

Myasthenia gravis (MG) is a prototypical antibody-mediated neurological autoimmune disease with the involvement of humoral immune responses in its pathogenesis. T follicular helper (Tfh) cells have been implicated in many autoimmune diseases. However, whether and how Tfh cells are involved in MG remain unclear. Here, we established and studied a widely-used and approved animal model of human MG, the rat model with acetylcholine receptor alpha (AChRα) subunit (R-AChR)-induced experimental autoimmune myasthenia gravis (EAMG). This model presented mild body-weight loss 10 days after the first immunization (representing the early stage of disease) and more obvious clinical manifestations and body-weight loss 7 days after the second immunization (representing the late stage of disease). AChR-specific pre-Tfh cells and mature Tfh cells were detected in these two stages, respectively. In co-cultures of Tfh cells and B cells, the number of IgG2b-secreting B cells and the level of anti-AChR antibodies in the supernatant were higher in the cultures containing EAMG-derived Tfh cells. In immunohistochemistry and immunofluorescence assays, a substantial number of CD4/Bcl-6 T cells and a greater number of larger germinal centers were observed in lymph node tissues resected from EAMG rats. Based on these results, we hypothesize that an AChR-specific Tfh cell-mediated humoral immune response contributes to the development of EAMG.

Effects of Yangyin Recipe on imbalance of Th17 and Treg cells in ApoE-/- mice under HSP65 attack / 中草药

Objective To investigate the effects of Yangyin Recipe on atherosclerosis (AS) lesions, helper T cells 17 (Th17), and regulatory T cells (Treg) in apolipoprotein E knockout (ApoE-/-) mice under the heat shock protein (HSP) 65 attack, and explore the potential mechanisms. Methods ApoE-/- mice were fed with normal and high-fat diets respectively, and injected with sterile PBS or HSP65. Simvastatin was used as the positive drug to study the effects of Yangyin Recipe on blood lipids, plaques, Th17/Treg cells and related inflammation indicators in AS model mice. Drugs were all intragastric administrated with equal volume of normal saline as a control. Serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) were detected by esterase method; The area of aortic plaque was determined by oil red O staining; The amounts of IL-17A+CD4+ Th17 and Foxp3+CD25+CD4+ Treg cells in anticoagulated blood were determined by flow cytometry; Serum IL-6, IL-17, IL-2, TGF-β, and IL-10 were determined by ELISA; mRNA expression of Stat3, Stat5a, Foxp3 in spleens and Stat3, Stat5a in aortas were detected by qPCR; Protein expression of phosphorylated STAT3 (pSTAT3), phosphorylated STAT5 (pSTAT5), FOXP3 in spleen, and pSTAT3 in aortas were determined by Western blotting. Results Under the influence of Yangyin Recipe, serum TC, TG, LDL levels decreased (P < 0.01), and HDL increased (P < 0.05); Aortic plaque area decreased remarkably (P < 0.01); IL-17A+CD4+Th17 cells in peripheral blood decreased (P < 0.05), Foxp3+CD25+CD4+ Treg increased (P < 0.05), and Th17/Treg ratio decreased drastically (P < 0.01); Serum IL-6 decreased (P < 0.05), IL-2 (P < 0.01), TGF-β and IL-10 (P < 0.05) increased; Protein levels of pSTAT3 in spleens decreased (P < 0.01), but pSTAT5 (P < 0.01) and FOXP3 (P < 0.05) increased; Aortic Stat3 mRNA (P < 0.01) and pSTAT3 (P < 0.05) protein decreased. Conclusion Yangyin Recipe can improve hyperlipidemia and AS in ApoE-/- mice during HSP65 attack, and regulate the balance of Th17/Treg in peripheral blood and the secretion of inflammatory factors to improve inflammation, which may be related to attenuate IL-6/pSTAT3 and enhance IL-2/pSTAT5 signaling.

Expression of follicular helper T cells in peripheral blood of patients with hepatic echinococcosis / 中国血吸虫病防治杂志

Objective To detect the expression of follicuLar helper T cells (Tfh) and interleukin-21 (IL-21) in the peripheral blood of patients with hepatic echinococcosis and healthy controls, so as to explore the associations of Tfh and IL-21 expression with the progression of hepatic echinococcosis. Methods Fifty cases of hepatic echinococcosis and healthy controls were collected from Qinghai Provincial People's Hospital, respectively. Flow cytometry was used to detect the expression of Tfh cells in the peripheral blood of hepatic echinococcosis patients and healthy controls, and enzyme-linked immunosorbent assay (ELISA) was used to detect serum IL-21 expression in hepatic echinococcosis patients and healthy controls. The correlation between Tfh cell expression and serum IL-21 level was examined in the patients with hepatic echinococcosis. Results Flow cytometry detected a higher percentage of CD4+CXCR5+ T cells (18.49% ± 5.67% vs. 16.18% ± 4.04%, P < 0.05), CD4+CXCR5+PD-1+ T cells (4.94% ± 1.91% vs. 2.29% ± 0.79%, P < 0.05) and CD4+CXCR5+ICOS+PD-1+ T cells (30.93% ± 24.10% vs. 21.07% ± 14.25%, P < 0.05) in hepatic echinococcosis patients than in healthy controls, and no significant difference was seen in the percentage of CD4+CRCR5+ICOS+ T cells between the patients and controls (0.29% ± 0.32% vs. 0.25% ± 0.31%, P > 0.05) . The serum IL-21 level was significantly higher in the patients with hepatic echinococcosis than in healthy controls ([ 293.35 ± 2 03.65) pg/mL vs. (192.72 ± 70.09) pg/mL, P < 0.05]; however, there was no correlation between the Tfh cell expression and serum IL-21 level in patients with hepatic echinococcosis (P > 0.05). Conclusion The expression of peripheral blood Tfh cells and serum IL-21 is elevated in patients with hepatic echinococcosis, and Tfh cells and IL-21 may contribute to the progression of hepatic echinococcosis.

AKT/mTOR/STATs signaling pathway regulates various immune cells that mediate MS / 医学研究生学报

AKT/mTOR/STATs signaling pathway not only plays an important role in tumor growth, but also in the regulation of immune system. Activated AKT promotes the activation of downstream signaling pathways mTORC1 and mTORC2 through phosphorylation. mTOR is currently being considered as an important regulator of immune system and plays an important role in regulating the function and metabolism of various immune cells. Multiple sclerosis (MS) is an autoimmune deficiency disease whose pathogenesis has not been fully elucidated. This article focuses on the regulation of AKT/mTOR/STATs signaling pathways in various immune cells such as macrophage M1/M2 polarization, B cell proliferation and differentiation, helper T lymphocyte (Th cell) proliferation and differentiation, and Treg cell proliferation and differentiation, which would be helpful to illustrate the role of the AKT/mTOR/STATs signaling pathway in mediating the regulation of immune cells in MS.