• Molecules and Cells
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• 제43권5호
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• pp.438-447
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• 2020

The aim of this study was to explore the role of IL-6-miR-210 in the regulation of Tregs function and atrial fibrosis in atrial fibrillation (AF). The levels of interleukin (IL)-6 and IL-10 in AF patients were detected by using ELISA. Proportions of Treg cells were detected by fluorescence activated cell sorting analysis in AF patients. The expression of Foxp3, α-SMA, collagen I and collagen III were determined by western blot. The atrial mechanocytes were authenticated by vimentin immunostaining. The expression of miR-210 was performed by quantitative real-time polymerase chain reaction (qRT-PCR). TargetScan was used to predict potential targets of miR-210. The cardiomyocyte transverse sections in AF model group were observed by H&E staining. The myocardial filaments were observed by masson staining. The level of IL-6 was highly increased while the level of IL-10 (Tregs) was significantly decreased in AF patients as compared to normal control subjects, and IL-6 suppressed Tregs function and promoted the expression of α-SMA, collagen I and collagen III. Furthermore, miR-210 regulated Tregs function by targeting Foxp3, and IL-6 promoted expression of miR-210 via regulating hypoxia inducible factor-1α (HIF-1α). IL-6-miR-210 suppresses regulatory T cell function and promotes atrial fibrosis by targeting Foxp3.

• Journal of Korean Medical Science
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• 제33권52호
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• pp.336.1-336.12
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• 2018

Background: We aimed to investigate mucosal immunity related to forkhead box P3 ($FOXP3^+$) regulatory T (Treg) cells, T helper 17 (Th17) cells and cytokines in pediatric inflammatory bowel disease (IBD). Methods: Mucosal tissues from terminal ileum and colon and serum samples were collected from twelve children with IBD and seven control children. Immunohistochemical staining was done using anti-human FOXP3 and anti-$ROR{\gamma}t$ antibodies. Serum levels of cytokines were analyzed using a multiplex assay covering interleukin $(IL)-1{\beta}$, IL-4, IL-6, IL-10, IL-17A/F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon $(IFN)-{\gamma}$, soluble CD40L, and tumor necrosis factor-${\alpha}$. Results: $FOXP3^+$ Treg cells in the lamina propria (LP) of terminal ileum of patients with Crohn's disease were significantly (P < 0.05) higher than those in the healthy controls. $ROR{\gamma}t^+$ T cells of terminal ileum tended to be higher in Crohn's disease than those in the control. In the multiplex assay, serum concentrations (pg/mL) of IL-4 ($9.6{\pm}1.5$ vs. $12.7{\pm}3.0$), IL-21 ($14.9{\pm}1.5$ vs. $26.4{\pm}9.1$), IL-33 ($14.3{\pm}0.9$ vs. $19.1{\pm}5.3$), and $IFN-{\gamma}$ ($15.2{\pm}5.9$ vs. $50.2{\pm}42.4$) were significantly lower in Crohn's disease than those in the control group. However, serum concentration of IL-6 ($119.1{\pm}79.6$ vs. $52.9{\pm}39.1$) was higher in Crohn's disease than that in the control. Serum concentrations of IL-17A ($64.2{\pm}17.2$ vs. $28.3{\pm}10.0$) and IL-22 ($37.5{\pm}8.8$ vs. $27.2{\pm}3.7$) were significantly higher in ulcerative colitis than those in Crohn's disease. Conclusion: Mucosal immunity analysis showed increased $FOXP3^+$ T reg cells in the LP with Crohn's disease while Th17 cell polarizing and signature cytokines were decreased in the serum samples of Crohn's disease but increased in ulcerative colitis.

• 한국미생물·생명공학회지
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• 제37권2호
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• pp.160-169
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• 2009

$\gamma$-PGA는 우리 전통 콩 발효식품인 청국장의 끈적끈적한 점액성의 성분으로, 매우 다양한 기능을 가지고 있는 천연 소재이다. 이러한 $\gamma$-PGA가 아토피발진 억제 가능성을 알아보기 위해 NC/Nga 생쥐를 사용하여 in vitro 실험을 실시하였다. $\gamma$-PGA(PGA-HM, 분자량 300 kDa)를 초음파처리로 저분자화시킨 30 kDa 이하의 저분자 PGA-LM를 만들고, 고분자 PGA-HM과 PGA-LM을 사용하여 실험하였는데 동일한 결과를 얻어 PGA-LM 실험결과 중심으로 보고한 것이다. 아토피 피부발진 NC/Nga 생쥐의 비장에서 B 세포와 T세포를 순수 분리하여 항알레르기 작용에 대한 in vitro 실험을 실시하였다. PGA-LM은 hFCs에 대한 세포독성 실험에서 모든 농도에서 세포독성을 나타내지 않았다. PGA-LM이 B 세포 분화 및 활성화에 미치는 영향을 관찰하기 위하여, NC/Nga 생쥐의 비장에서 순수 분리한 B 세포에 anti-CD40/rmIL-4로 자극한 결과, 대조군은 전사인자인 NF-${\kappa}B$의 활성화로 IL-$1\beta$, IL-6, 그리고 TNF-$\alpha$ mRNA의 발현이 증가되었다. 그러나 PGA-LM과 양성대조군인 rmIL-10 투여군은 염증사이토카인 IL-$1\beta$, IL-6 그리고 TNF-$\alpha$ mRNA 유전자 발현이 감소하였고, IL-10 mRNA 유전자 발현은 증가하였으나 TGF-$\beta$ mRNA의 유전자 발현은 대조군과 큰 차이가 나타나질 않았다. 또한 CD4+ T 세포에 PGA-LM $100\;{\mu}g/ml$를 처리한 후 4일간 동시 배양하여 CD4+IFN-$\gamma$+와 CD4+CD25+foxp3+ Treg 세포를 세포내 염색으로 분석한 결과에서는 CD4+IFN-$\gamma$+인 Th1 세포의 증가와 CD4+CD25+foxp3+ Treg 세포를 증가시켜 알레르기반응에서 우위한 Th2 세포에서 Th1 세포로 전환시키는 면역조절 역할을 나타내었다. 이상의 결과로 NC/Nga 생쥐에서 PGA-LM은 염증유전자 발현을 억제시키고 IFN-$\gamma$+의 증가 및 조절 T 세포의 유도로 아토피피부염의 피부발진을 치료하는 면역조절제로 사용될 수 있을 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제54권2호
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• pp.163-171
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• 2016

As most infections by the helminth parasite elicit the recruitment of $CD4^+CD25^+Foxp3^+$ T ($T_{reg}$) cells, many scientists have suggested that these cells could be used for the treatment of immune-mediated inflammation and associated diseases. In order to investigate the distribution and alteration of activated $T_{reg}$ cells, we compared the expression levels of $T_{reg}$ cell activation markers in the ileum and gastrocnemius tissues 1, 2, and 4 weeks after infection. The number of $T_{reg}$ cells was monitored using GFP-coded Foxp3 transgenic mice. In mice at 1 week after Trichinella spiralis infection, the number of activated $T_{reg}$ cells was higher than in the control group. In mice at 2 weeks after infection, there was a significant increase in the number of cells expressing Foxp3 and CTLA-4 when compared to the control group and mice at 1 week after infection. At 4 weeks after infection, T. spiralis was easily identifiable in nurse cells in mouse muscles. In the intestine, the expression of Gzmb and Klrg1 decreased over time and that of Capg remained unchanged for the first and second week, then decreased in the 4th week. However, in the muscles, the expression of most chemokine genes was increased due to T. spiralis infection, in particular the expression levels of Gzmb, OX40, and CTLA-4 increased until week 4. In addition, increased gene expression of all chemokine receptors in muscle, CXCR3, CCR4, CCR5, CCR9, and CCR10, was observed up until the 4th week. In conclusion, various chemokine receptors showed increased expressions combined with recruitment of $T_{reg}$ cells in the muscle tissue.

• Parasites, Hosts and Diseases
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• 제50권1호
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• pp.29-35
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• 2012

The aim of the study is to characterize the phenotypes of $CD4^+$ $CD25^+$ T regulatory cells within the liver granulomas and association with both Foxp-3 gene expression and splenic cytokines. Naive C57BL/6 mice were intravenously injected with multiple doses of the soluble egg antigen (SEA) 7 days before cercarial infection. The immunized and infected control groups were sacrificed 8 and 16 weeks post-infection (PI). Histopathology, parasitological parameters, splenic phenotypes for T regulatory cells, the FOXP-3 expression in hepatic granuloma using real-time PCR, and the associated splenic cytokines were studied. Histopathological examination of the liver revealed remarkable increase in degenerated ova within hepatic granuloma which decreased in diameter at weeks 8 and 16 PI ($P$<0.01). The percentage of T regulatory cells ($CD4^+$ $CD25^+$) increased significantly ($P$<0.01) in the immunized group compared to the infected control at weeks 8 and 16 PI. The FOXP-3 expression in hepatic granulomas increased from 10 at week 8 to 30 fold at week 16 PI in the infected control group. However, its expression in the immunized group showed an increase from 30 at week 8 to 70 fold at week 16 PI. The splenic cytokine levels of pro-inflammatory cytokines, IFN-${\gamma}$, IL-4, and TNF-${\alpha}$, showed significant decreases ($P$<0.05) compared to the infected control group. In conclusion, the magnitude and phenotype of the egg-induced effects on T helper responses were found to be controlled by a parallel response within the T regulatory population which provides protection in worm parasite-induced immunopathology.

• Biomolecules & Therapeutics
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• 제25권2호
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• pp.130-139
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• 2017

$CXCR5^+$ T follicular helper (Tfh) cells are associated with aberrant autoantibody production in patients with antibody-mediated autoimmune diseases including lupus. Follicular regulatory T (Tfr) cells expressing CXCR5 and Bcl6 have been recently identified as a specialized subset of $Foxp3^+$ regulatory T (Treg) cells that control germinal center reactions. In this study, we show that retroviral transduction of CXCR5 gene in $Foxp3^+$ Treg cells induced a stable expression of functional CXCR5 on their surface. The Cxcr5-transduced Treg cells maintained the expression of Treg cell signature genes and the suppressive activity. The expression of CXCR5 as well as Foxp3 in the transduced Treg cells appeared to be stable in vivo in an adoptive transfer experiment. Moreover, Cxcr5-transduced Treg cells preferentially migrated toward the CXCL13 gradient, leading to an effective suppression of antibody production from B cells stimulated with Tfh cells. Therefore, our results demonstrate that enforced expression of CXCR5 onto Treg cells efficiently induces Tfr cell-like properties, which might be a promising cellular therapeutic approach for the treatment of antibody-mediated autoimmune diseases.

• Journal of Gastric Cancer
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• 제20권2호
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• pp.190-201
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• 2020

Purpose: This study sought to investigate the prognostic significance of tumor-infiltrating lymphocytes (TILs) in relation to tumor location within the stomach. Materials and Methods: The densities and prognostic significance of TIL subsets were evaluated in 542 gastric cancer patients who underwent gastrectomy. Immunohistochemical staining for CD3, CD4, CD8, forkhead/winged helix transcription factor (Foxp3), and granzyme B was performed. Results: Cardia cancer was associated with significantly lower densities of CD8 T-cells and higher densities of Foxp3 and granzyme B T-cells than non-cardia tumors. Multivariate analysis showed that advanced age (hazard ratio [HR], 1.023; 95% confidence interval [CI], 1.006-1.040), advanced T classification (HR, 2.029; 95% CI, 1.106-3.721), lymph node metastasis (HR, 3.319; 95% CI, 1.947-5.658), low CD3 expression (HR, 0.997; 95% CI, 0.994-0.999), and a high Foxp3/CD4 ratio (HR, 1.007; 95% CI, 1.001-1.012) were independent predictors of poor overall survival in cardia cancer patients. In non-cardia cancer patients, total gastrectomy (HR, 2.147; 95% CI, 1.507-3.059), advanced T classification (HR, 2.158; 95% CI, 1.425-3.266), lymph node metastasis (HR, 1.854; 95% CI, 1.250-2.750), and a low Foxp3/CD4 ratio (HR, 0.978; 95% CI, 0.959-0.997) were poor prognostic factors for survival. Conclusions: The densities and prognostic effects of TILs differed in relation to the location of tumors within the stomach. The contrasting prognostic effects of Foxp3/CD4 ratio in cardia and non-cardia gastric cancer patients suggests that clinicians ought to consider tumor location when determining treatment strategies.

• 한국미생물·생명공학회지
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• 제38권1호
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• pp.53-63
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• 2010

$\gamma$-PGA는 우리 전통 콩 발효식품인 청국장의 끈적끈적한 점액성 성분의 하나로, 매우 다양한 기능을 가지고 있는 천연물질이다. 이러한 $\gamma$-PGA의 아토피발진 억제 효과를 알아보고자 NC/Nga 생쥐를 사용하여 in vivo 실험을 하였다. BMAC로 유도된 NC/Nga 아토피 피부발진 생쥐에 분자량 300 kDa인 $\gamma$-PGA(PGA-HM)와 이를 저분자화 시킨 저분자 $\gamma$-PGA(PGA-LM)을 경구 투여한 결과 PGA-LM 투여군에서 clinical skin severity score가 유의성 있게 감소하였다. 혈청 IgE 수준은 PGA-LM이 대조군에 비하여 유의적으로 감소하였고, 혈청 IgG1 수준은 대조군에 비하여 감소하였으나 두 군 모두 유의성이 없었다. $CD4^+CD25^+foxp3^+$ Treg 세포가 유도되는 것을 확인하기 위하여, PGA-HM와 PGA-LM를 투여한 NC/Nga 아토피 피부발진 생쥐의 등 부위에서 mRNA를 분리히여 real-time PCR로 foxp3 mRNA 유전자 발현량을 측정한 결과는 대조군에 비하여 PGA-LM 투여군이 약 2배 이상 증가를 나타내었다. 또한 등피부 조직의 조직검사에서도 epidermis의 두께, 비만세포 침윤, 그리고 $CCR3^+$ 세포수 등이 대조군에 비하여 현저히 억제됨을 관찰할 수 있었다. 이상의 결과로 PGA-HM보다 PGA-LM이 BMAC로 아토피 피부염이 유발된 NC/Nga 생쥐에 $CD4^+CD25^+foxp3^+$ Treg 세포를 활성화하여 IgE 및 염증 사이토카인의 생산 및 $CCR3^+$ 호산구의 활성화가 억제되어 아토피조절 효과를 나타내는 것으로 사료되었다.

• Parasites, Hosts and Diseases
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• 제51권3호
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• pp.289-295
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• 2013

Different functions have been attributed to $CD4^+CD25^+Foxp3^+$ regulatory T-cells (Tregs) during malaria infection. Herein, we describe the disparity in Treg response and pro- and anti-inflammatory cytokines during infection with Plasmodium berghei ANKA between young (3-week-old) and middle-aged (8-month-old) C57BL/6 mice. Young mice were susceptible to cerebral malaria (CM), while the middle-aged mice were resistant to CM and succumbed to hyperparasitemia and severe anemia. The levels of pro-inflammatory cytokines, such as TNF-${\alpha}$, in young CM-susceptible mice were markedly higher than in middle-aged CM-resistant mice. An increased absolute number of Tregs 3-5 days post-inoculation, co-occurring with elevated IL-10 levels, was observed in middle-aged CM-resistant mice but not in young CM-susceptible mice. Our findings suggest that Treg proliferation might be associated with the suppression of excessive pro-inflammatory Th1 response during early malaria infection, leading to resistance to CM in the middle-aged mice, possibly in an IL-10-dependent manner.

• Journal of Gastric Cancer
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• 제17권4호
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• pp.384-393
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• 2017

Purpose: The tumor microenvironment is known to be associated with the metabolic activity of cancer cells and local immune reactions. We hypothesized that glucose metabolism measured by 2-deoxy-2-($^{18}F$)fluoro-D-glucose ($^{18}F-FDG$) positron emission tomography (PET)-computed tomography (CT) ($^{18}F-FDG$ PET-CT) would be associated with local immune responses evaluated according to the presence of tumor infiltrating lymphocytes (TILs). Materials and Methods: We retrospectively reviewed 56 patients who underwent $^{18}F-FDG$ PET-CT prior to gastrectomy. In resected tumor specimens, TIL subsets, including cluster of differentiation (CD) 3, CD4, CD8, Forkhead box P3 (Foxp3), and granzyme B, were subjected to immunohistochemical analysis. The prognostic nutritional index (PNI) was calculated as: ($10{\times}serum$ albumin value)+($0.005{\times}peripheral$ lymphocyte counts). Additionally, the maximum standard uptake value ($SUV_{max}$) was calculated to evaluate the metabolic activity of cancer cells. Results: The $SUV_{max}$ was positively correlated with larger tumor size (R=0.293; P=0.029) and negatively correlated with PNI (R=-0.407; P=0.002). A higher $SUV_{max}$ showed a marginal association with higher CD3 (+) T lymphocyte counts (R=0.227; P=0.092) and a significant association with higher Foxp3 (+) T lymphocyte counts (R=0.431; P=0.009). No other clinicopathological characteristics were associated with $SUV_{max}$ or TILs. Survival analysis, however, indicated that neither $SUV_{max}$ nor Foxp3 held prognostic significance. Conclusions: FDG uptake on PET-CT could be associated with TILs, especially regulatory T cells, in gastric cancer. This finding may suggest that PET-CT could be of use as a non-invasive tool for monitoring the tumor microenvironment in patients with gastric cancer.