The value of peripheral blood and pleural effusion T-SPOT.TB combined with pleural effusion adenosine deaminase in diagnosis of tuberculous pleurisy / 中国热带医学

@#Abstract: Objective To explore the relationship between peripheral blood and pleural effusion tuberculosis (TB) infection effector T cells, and to further evaluate the value of combined pleural effusion adenosine deaminase (ADA) for rapid diagnosis of tuberculous pleurisy. Methods The test data of 80 cases of tuberculous pleurisy and 70 cases of nontuberculous pleurisy treated in the Sixth People's Hospital of Nantong City from January 2017 to December 2020 were analyzed. The TBinfected effector T cells were also detected simultaneously in the peripheral blood and the pleural effusion by the T-SPOT technique, and the pleural effusion ADA was detected by the rate method. The subject operating characteristic curve (ROC) was applied to take the optimal pleural effusion ADA threshold to compare the sensitivity and specificity of different critical values. Person phase analysis was applied to analyze the correlation between peripheral blood and pleural effusion T-SPOT.TB. Data of peripheral blood, pleural effusion T-SPOT.TB and ADA were integrated. Results When pleural effusion ADA>45 U/L, the sensitivity and specificity for the diagnosis of tuberculous pleurisy were 50.0% and 94.3%, respectively; when ADA > 25.15 U/ L, the sensitivity and specificity were 80.0% and 72.9%. When ADA > 45 U / L, pleural/ blood T-SPOT.TB spot ratio (spot forming cells, SFCs) > 2 times, the specificity for the diagnosis of tuberculous pleurisy was 100% (highest); when 25.15 U/L< pleural effusion ADA ≤ 45 U/L, pleural/blood T-SPOT.TB spot ratio > 2 times, the specificity for the diagnosis of tuberculous pleurisy was 92.3% (second). When pleural effusion ADA ≤ 25.15 U/L, and the pleural effusion/blood T-SPOT.TB spot number ratio > 2 times, with 83.3% specificity (the lowest of the three groups). Conclusions The level of pleural effusion ADA is one of the most used methods for diagnosing tuberculous pleurisy. Further combination of pleural effusion and blood T-SPOT.TB, if the ratio of pleural effusion / blood T-SPOT. TB spots is greater than 2 times, it can further improve the diagnosis rate of tuberculous pleurisy.

Effect of metformin on bleomycin-induced systemic sclerosis mice by regulating the balance of Teff/regulatory T cells / 中华风湿病学杂志

Objective:To explore the effect and mechanism of different concentrations of metformin on bleomycin (BLM)-induced systemic sclerosis (SSc) mice model.Methods:C57BL/6 mice were divided into the normal group, the model group, the high, the medium and the low metformin (MET) treatment groups randomly. All mice were sacrificed after BLM and metformin treatment for 4 weeks. Local skin was exminedby histopathological staining method to measure the thickness of dermis and collagen, and immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) were used to detect the protein and mRNA levels of Interleukin (IL)-17, forkhead box P3 (Foxp3) and α-smooth muscle actin (α-SMA). Flow cytometry was used to detect the percentage of effector T cell (Teff) and regulatory cells (Treg) in splenic mononuclear cells. The data such as dermal collagen thickness, α-SMA, IL-17, Foxp3, Teff and Treg levels were statistically analyzed by one-way analysis of variance. The data such as dermal collagen thickness, α-SMA, IL-17, Foxp3, Teff and Treg levels were analyzed by one-way analysis of variance, and least significant difference (LSD)- t or Kruskal-Wallis test was used for comparison between groups. Results:Compared with the normal group, remarkable fibrotic lesions appeared in the skin of mice in the model group, and the levels of T-helper cells (Th)1, Th2, Th17, and T follicular helper cells (Tfh) cells were increased, accompanied by a significant decrease in the level of Treg cells. After high-dose metformin treatment, the dermal thickness [(131±25) μm], collagen thickness [(119±18) μm], and α-SMA [(3.0±0.5)/HPH] were significantly reduced( F=14.390, P<0.01; F=40.245, P<0.01; F=44.626, P<0.01). Th1[(27.00±6.68)%], Th17[(0.56±0.20)%], Tfh[(6.4±1.6)%] cells ware significantlyreduced ( F=32.390, P<0.01; F=16.083, P<0.01; F=16.546, P<0.01), and Treg[(11.23±1.52)%] cells were significantly increased ( F=10.171, P<0.01). Conclusion:Metformin can effectively reverse the local skin changesin BLM-induced SSc mouse model, and show immune regulation and anti-fibrosis effects by restoring the Teff/Treg balance.

Differences in susceptibility to Lewis lung carcinoma and T lymphocyte subsets of aged mice / 中华老年医学杂志

Objective To investigate the differences in susceptibility to Lewis lung carcinoma and T lymphocyte subsets in the immune microenvironment between young and elderly mice.Methods Six C57/B6 mice at two months(young)and six mice at twelve months(aged)were injected with Lewis lung carcinoma cells at the dose of 1 × 106 in the left armpit to establish a murine model of lung carcinoma.The weight and tumor growth were monitored.Blood samples for routine blood tests were collected after 24 days.The proportions of CD4+ and CD8+T cells in the spleen were detected by flow cytometry,and the infiltration of CD4+,CD8+ T cells and related effector T cells in the tumor microenvironment were determined in the same way.Results The body weight of tumor bearing mice in the aged group was significantly higher than that in the young group(P ＜0.001);The tumor weight in the aged group(5.084±0.528)g was significantly higher than that in the young group(2.963 ±0.378)g(t =3,349,P =0.012);Routine blood tests showed that the numbers of leukocytes and subsets(except mononuclear)in the aged group were significantly lower than in the young group(P ＜0.05);Flow cytometry found that the effector and memory/effector CD4+T cell ratios in the spleen were significantly higher in the aged group than in the young group(P ＜0.001)and the expression of effector and memory/effector CD8+T cells in the tumor microenvironment was also significantly higher than in the young group(P ＜0.05);Quantitative expression values of IL-6 and IL-10 in the tumor microenvironment were 25090±3820 and 10670± 1793 in the aged group and 6252±864 and 3061±451 in the young group,respectively.Moreover,the expression levels of IL-6 and IL-10(t =3.925,P =0.01;t =3.552,P =0.02)in the tumor microenvironment in the aged group were significantly lower than those in the young group.Conclusions Young mice are more susceptible to Lewis lung carcinoma,probably as a result of differences in inflammation and immunity.

Effect of pioglitazone on balance of regulatory and effector T cells of ure-mic apolipoprotein E knockout mice in vitro / 中国病理生理杂志

AIM:To investigate the effects of pioglitazone on the quantity and function-related factors of regu-latory and effector T cells ( Treg and Teff ) of uremic apolipoprotein E knockout mice in vitro with or without the stimulation of atherosclerotic plaque-specific antigen oxidized low-density lipoprotein ( oxLDL) .METHODS:Uremic apolipoprotein E knockout mouse model was established by 2-step surgical procedure.After intervention with different concentrations ( 2μmol/L and 20μmol/L) of pioglitazone and PPARγantagonist GW9662 (5μmol/L) on splenocytes of uremic mice for 12 h in the presence or absence of oxLDL (2 mg/L), the levels of CD4 +CD25 +Foxp3 +Treg and IFNγ+CD4 +Teff were de-termined by flow cytometry.The mRNA expressions of Foxp3 and IFNγwas detected by real-time fluorescent quantitative PCR.RESULTS:In vitro, oxLDL induced a Treg/Teff imbalance in splenocytes from the uremic mice.Pioglitazone up-regulated the level of Treg and mRNA expression of Foxp3 in the presence of oxLDL, which was not antagonized by GW9662.Meanwhile, pioglitazone downregulated the level of Teff and mRNA expression of IFNγin the presence or ab-sence of oxLDL, which was reversed by GW9662.CONCLUSION:oxLDL induces a Treg/Teff imbalance in uremic apo-lipoprotein E knockout mice.Pioglitazone modulates the Treg/Teff imbalance probably through PPARγ-independent and-dependent mechanisms.

Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

Regulatory T (TREG) cells play an important role in maintaining immune tolerance and avoiding autoimmunity. We analyzed the expression of membrane molecules in TREG and effector T cells in systemic lupus erythematosus (SLE). TREG and effector T cells were analyzed for the expression of CTLA-4, PD1, CD28, CD95, GITR, HLA-DR, OX40, CD40L, and CD45RO in 26 patients with active disease, 31 with inactive disease, and 26 healthy controls. TREG cells were defined as CD25+/highCD127Ø/lowFoxP3+, and effector T cells were defined as CD25+CD127+FoxP3Ø. The ratio of TREG to effector T cells expressing GITR, PD1, HLA-DR, OX40, CD40L, and CD45RO was determined in the three groups. The frequency of TREG cells was similar in patients with SLE and controls. However, SLE patients had a decreased frequency of CTLA-4+TREG and CD28+TREG cells and an increased frequency of CD40L+TREG cells. There was a decrease in the TREG/effector-T ratio for GITR+, HLA-DR+, OX40+, and CD45RO+ cells, and an increased ratio of TREG/effector-T CD40L+ cells in patients with SLE. In addition, CD40L+TREG cell frequency correlated with the SLE disease activity index (P=0.0163). In conclusion, our findings showed several abnormalities in the expression of functionally critical surface molecules in TREG and effector T cells in SLE that may be relevant to the pathogenesis of this disease.

Cytokine Production in Patients With Leprosy / 나학회지

No abstract available.

Effect of Xuebijing injection on lipopolysaccharide-induced apoptosis of CD4+CD25+regulatory T cells and immune function of effector T cells in vitro / 中华急诊医学杂志

Objective To investigate the effect of Xuebijing injectiong on lipopolysaccharide(LPS)-induced apoptosis of CD4+CD25+regulatory T cells(Tregs)and immune function of effector T cells(Teff)in vitro.Method CD4+CD25+Tregs isolated from rat spleens were divided into the control group,anti-CD3/CD28 group,anti-CD3/CD28+LPS group,anti-CD3/CD28+Xuebijing injection group,and anti-CD3/CD28+LPS+Xuebijing injection group.The apoptosis rate and expression of forkhead/winged helix transcription factor p3 (Foxp3)and cytotoxic T-lymphocyte-associated antigen 4(CTLA-4)of CD4+CD25+Tregs were detected by flow cytometry(FCM),and the secretion of IL-10 of Tregs was measured by ELBA on day 3.CD4+CD25-T cells were co-cultured with CD4+CD25+Tregs(1:1)for 68 hours,proliferative activity of Teff was determined by MTT,and interleukin(IL)-2/sIL-2Rα levels were measured by ELISA.Results The apoptosis rate of CD4+CD25+Tregs in anti-CD3/CD28 group was 33.70± 3.06%,which was significantly higher than that in control group(12.84±0.84%).Also,apoptosis rate of CD4 CD25+Tregs in anti-CD3/CD28+LPS+Xuebijing injection group(45.13±2.70%)was much higher than that in anti-CD3/CD28+IPS group(29.41 ± 1.63%,P＜0.01).The expression of Foxp3 as well as CTLA-4,and the secretion of IL-10 were markedly decreased along with increases in the apoptosis rates.Compared with control group(54.48%),the mean inhibitory rate of Teff proliferative activity in response to Con A was significantly decreased in anti-CD3/CD28+Xuebijing injection group(31.26%,P＜0.05),and it was markedly decreased in anti-CD3/CD28+LPS+Xuebijing injection group comaped to anti-CD3/CD28+LPS group(P＜0.01).In addition,IL-2 levels in the supernatant of anti-CD3/CD28+Xuebijing injection group and anti-CD3/CD28+LPS+Xuebijing injection group were significantly higher than those of anti-CD3/CD28+IPS group(P＜0.01).Conclusions The inhibitory activity of CD4+CD25+Tregs on Teff appears to be upregulated by IPS stimulation in vitro,and Xuebijing injection could markedly enhance apoptosis of CD4+CD25+Tregs,thereby improving suppressive immune function of Teff.