regulatory t cells treg  (R&D Systems)

Journal: Frontiers in Neurology

Article Title: Serum retinol-binding protein 4 in stroke patients: correlation with T helper 17/regulatory T cell imbalance and 3-year cognitive function decline

doi: 10.3389/fneur.2023.1217979

Figure Lengend Snippet: Comparison of Th17 cells, Treg cells, and Th17/Treg ratios between stroke patients and HCs.

Article Snippet: Then, the proportions of Th17 and Treg cells in CD4 + T cells were determined by flow cytometry (FCM) using the FlowX Human Th17 Cell Multi-Color Flow Cytometry Kit (No. Cat. FMC007B, R&D Systems, Inc., Minneapolis, Minnesota, United States) and the Regulatory T Cell (Treg) Flow Cytometry Panel (No. Cat. FMC-P-004, R&D Systems, Inc., Minneapolis, Minnesota, USA).

Techniques: Comparison

Journal: Frontiers in Neurology

Article Title: Serum retinol-binding protein 4 in stroke patients: correlation with T helper 17/regulatory T cell imbalance and 3-year cognitive function decline

doi: 10.3389/fneur.2023.1217979

Figure Lengend Snippet: Serum RBP4 was positively associated with a Th17/Treg imbalance. Association of serum RBP4 with Th17 cells (A) , Treg cells (B) , and the Th17/Treg ratio (C) in stroke patients. Association of serum RBP4 with Th17 cells (D) , Treg cells (E) , and the Th17/Treg ratio (F) in HCs.

Article Snippet: Then, the proportions of Th17 and Treg cells in CD4 + T cells were determined by flow cytometry (FCM) using the FlowX Human Th17 Cell Multi-Color Flow Cytometry Kit (No. Cat. FMC007B, R&D Systems, Inc., Minneapolis, Minnesota, United States) and the Regulatory T Cell (Treg) Flow Cytometry Panel (No. Cat. FMC-P-004, R&D Systems, Inc., Minneapolis, Minnesota, USA).

Techniques:

Journal: Frontiers in Neurology

Article Title: Serum retinol-binding protein 4 in stroke patients: correlation with T helper 17/regulatory T cell imbalance and 3-year cognitive function decline

doi: 10.3389/fneur.2023.1217979

Figure Lengend Snippet: Serum RBP4 and Th17/Treg imbalances were negatively associated with the MMSE score at enrollment. Association of serum RBP4 (A) , Th17 cells (B) , Treg cells (C) , and the Th17/Treg ratio (D) with MMSE score at enrollment in stroke patients.

Article Snippet: Then, the proportions of Th17 and Treg cells in CD4 + T cells were determined by flow cytometry (FCM) using the FlowX Human Th17 Cell Multi-Color Flow Cytometry Kit (No. Cat. FMC007B, R&D Systems, Inc., Minneapolis, Minnesota, United States) and the Regulatory T Cell (Treg) Flow Cytometry Panel (No. Cat. FMC-P-004, R&D Systems, Inc., Minneapolis, Minnesota, USA).

Techniques:

Journal: Frontiers in Neurology

Article Title: Serum retinol-binding protein 4 in stroke patients: correlation with T helper 17/regulatory T cell imbalance and 3-year cognitive function decline

doi: 10.3389/fneur.2023.1217979

Figure Lengend Snippet: Serum RBP4 and Th17/Treg imbalances were associated with the occurrence of cognitive impairment at enrollment. Association of serum RBP4 (A) , Th17 cells (B) , Treg cells (C) , and the Th17/Treg ratio (D) with cognitive impairment at enrollment in stroke patients.

Article Snippet: Then, the proportions of Th17 and Treg cells in CD4 + T cells were determined by flow cytometry (FCM) using the FlowX Human Th17 Cell Multi-Color Flow Cytometry Kit (No. Cat. FMC007B, R&D Systems, Inc., Minneapolis, Minnesota, United States) and the Regulatory T Cell (Treg) Flow Cytometry Panel (No. Cat. FMC-P-004, R&D Systems, Inc., Minneapolis, Minnesota, USA).

Techniques:

Journal: Cells

Article Title: Propranolol Suppresses the T-Helper Cell Depletion-Related Immune Dysfunction in Cirrhotic Mice

doi: 10.3390/cells9030604

Figure Lengend Snippet: Serum T-helper 1 (Th1)/Treg cytokines were increased and serum Th2 cytokines were decreased in cirrhotic mice. Serum levels of Th1 ( A ), Th2 ( B ), and Treg ( C ) cytokines. * p < 0.05 vs. sham-V group; † p < 0.05 vs. BDL-V/TAA-V group.

Article Snippet: They were evaluated for plasma norepinephrine levels (Lifespan BioSciences, Seattle, WA, USA), levels of peripheral Th1 (IFN-γ and TNF-α), Th2 (IL-4 and IL-10), and Treg (TGFβ and IL-35) cytokines [mouse Quantikine ELISA kits (R&D Systems, Minneapolis, MN, USA)].

Techniques:

Journal: Cells

Article Title: Propranolol Suppresses the T-Helper Cell Depletion-Related Immune Dysfunction in Cirrhotic Mice

doi: 10.3390/cells9030604

Figure Lengend Snippet: High tissue levels of ADRB1/ADRB2 proteins were associated with high levels of Th1/Treg cytokines and LBP in the spleens of cirrhotic mice. ( A – C ) levels of ADRB1/ADRB2 proteins and of Th1/Th2/Treg cytokines; ( D – F ) Mean levels of INF-γ, IL-10 and IL-35 in cirrhotic mice of “high” ADRB and “low” ADRB groups. ( G ) Splenic LBP level between groups; ( H ) LBP levels in cirrhotic mice of “high” ADRB group and “low” ADRB group. ( I ) ADRB protein Levels between “high LBP” and “low LBP” groups. In cirrhotic mice, “high” ADRB or LBP groups were defined by high percentages (> third percentile, cut-off point is 393 for ADRB protein levels, and 1057 for splenic LBP levels) of summative increased splenic ADRB1 and ADRB2 protein or LBP levels compared to sham-V mice. * p < 0.05 vs. sham-V/high ADRB or LBP groups; † p < 0.05 vs. BDL-V/TAA-V group; LBP: lipopolysaccharide binding protein.

Article Snippet: They were evaluated for plasma norepinephrine levels (Lifespan BioSciences, Seattle, WA, USA), levels of peripheral Th1 (IFN-γ and TNF-α), Th2 (IL-4 and IL-10), and Treg (TGFβ and IL-35) cytokines [mouse Quantikine ELISA kits (R&D Systems, Minneapolis, MN, USA)].

Techniques: Binding Assay

Journal: Cells

Article Title: Propranolol Suppresses the T-Helper Cell Depletion-Related Immune Dysfunction in Cirrhotic Mice

doi: 10.3390/cells9030604

Figure Lengend Snippet: Acute propranolol incubation inhibited Treg-conditioned differentiation and restored Th2-conditioned differentiation of Th cells isolated from the spleens of cirrhotic mice. ( A , B ) Representative images and bar graphs of proteins and mRNAs (%/18S); ( C ) Expressions in cell lysates of Th cells isolated from spleen of sham-V or TAA-V mice; ( D ) Flow cytometry-based intracellular cytokine staining for percentages of Th1/Th2/Treg-conditioned differentiation among groups. * p < 0.05 vs. sham-V group; † p < 0.05 vs. TAA-V group.

Article Snippet: They were evaluated for plasma norepinephrine levels (Lifespan BioSciences, Seattle, WA, USA), levels of peripheral Th1 (IFN-γ and TNF-α), Th2 (IL-4 and IL-10), and Treg (TGFβ and IL-35) cytokines [mouse Quantikine ELISA kits (R&D Systems, Minneapolis, MN, USA)].

Techniques: Incubation, Isolation, Flow Cytometry, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: NLRP6 Deficiency in CD4 T Cells Decreases T Cell Survival Associated with Increased Cell Death.

doi: 10.4049/jimmunol.1800938

Figure Lengend Snippet: FIGURE 2. Reduced IFN-g by Nlrp6-deficient Th1 cells. (A) Naive CD4+ T cells from wt or Nlrp6- deficient mice were differentiated into Th1 cells, and IFN-g–producing cells were measured by flow cytom- etry on day 5. (B) The percentage and (C) numbers of IFN-g–producing wt and Nlrp6-deficient Th1 cells. Each data point represents one animal and the median is shown. The Mann–Whitney U test was used for data analysis. (D) IFN-g production was quantified by ELISA in wt and Nlrp6-deficient Th1 cells before and after PMA/ionomycin stimulation. Naive CD4+ T cells from wt (n = 3) or Nlrp6-deficient mice (n = 3) were differ- entiated for 5 d into Th1 cells. IFN-g–producing cells were determined every single day by flow cytometry. (E) Percentage and (F) numbers of IFN-g–producing T cells are presented. Data are mean and error bars represent SD. (G) Wt and Nlrp6-deficient naive CD4+ T cells were differentiated into Th2 cells for 5 d and into Th17 cells and Foxp3+ Treg cells for 4 d. IL-13– and IL-17A– producing cells following PMA/ionomycin stimulation and intracellular Foxp3+ were determined by flow cytometry. Respective dot plots were obtained after gating on viable single cells. Percentages of IL-13+, IL-17A+, or Foxp3+ T cells were indicated on respective gates. (H) Percentage and number of Th2, Th17, or Treg cells are presented. Each data point indicates an indi- vidual animal; results are shown as medians (n = 4 for each group). Data were analyzed with a Kruskal–Wallis test. *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: FOXP3-positive regulatory T (Treg) cells were differentiated in the presence of 10 mg/ml anti–IL-4 Ab, 10 mg/ml anti–IFN-g by guest on M ay 31, 2019 http://w w w .jim m unol.org/ D ow nloaded from Ab, 20 ng/ml IL-2 (R&D Systems), and 5 ng/ml hTGF-b (R&D Systems).

Techniques: MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Cytometry

Journal: Scientific Reports

Article Title: Inflammatory Responses Reprogram T REGS Through Impairment of Neuropilin-1

doi: 10.1038/s41598-019-46934-x

Figure Lengend Snippet: PMMA particles induce NF-κB activity and alter bone marrow cellularity toward reduced immunosuppression. ( a ) PMMA particles induced local NF-κB activation in immune cells. Total cells were isolated from the bone marrow of RelA-Luciferase reporter mice post PMMA intra-tibial injection. After removal of red blood cells, mononucleated cells from bone marrow of separated femurs and tibias were lysed to assess luciferase activity, which was normalized by protein concentration determined by standard BCA assay. ( b ) Increase of myeloid progenitor cells in the bone marrow of mice 2 days post intra-tibial injection of PMMA particles revealed by flow cytometric analysis. 1 × 10 7 mononucleated bone marrow cells were stained with FACS antibodies to assess myeloid progenitor populations including LSK HSCs, CMPs and GMPs. ( c ) Decreased frequency of bone marrow CD4 + CD25 + Foxp3 + T REG by intra-tibial PMMA injection. ( d ) Alterations of bone marrow myeloid lineages. Monocytic myeloid cells are marked as CD11b + Gr-1 − and granulocytic cells as CD11b + Gr-1 + . ( e , f ) Assessment of osteoclastogenic potential of whole bone marrow cells (WBM) by ex vivo osteoclastogenesis assay (OCgenesis) 2 days after intra-tibial injection of PMMA. 50,000, 100,000 or 200,000 total bone marrow cells were cultured in 96-well plates supplemented with CMG and RANKL to achieve optimal cell density for osteoclast formation. After 4 days of culture, cells were fixed before subjected TRAP staining to visualize osteoclasts. Cells with expanded cytoplasm and more than 3 nuclei were counted as matured osteoclasts. ( e ) Representative images and quantification of the number of multi-nucleated (MNC) osteoclasts per well counted from triplicates of three independent experiments. All columns in the graphs were represented as mean ± SD. *p < 0.05; **p < 0.005 or as indicated by Student T-test.

Article Snippet: To transduce T REGS , 1 millions of freshly MACS-isolated naïve CD4+ T cells from mouse spleen were incubated overnight with 3 ml of retroviral stock supplemented with reagents from the T REG Cell Differentiation Kit (R&D Systems) and 10ug/ml of polybrene.

Techniques: Activity Assay, Activation Assay, Isolation, Luciferase, Injection, Protein Concentration, BIA-KA, Staining, Ex Vivo, Cell Culture

Journal: Scientific Reports

Article Title: Inflammatory Responses Reprogram T REGS Through Impairment of Neuropilin-1

doi: 10.1038/s41598-019-46934-x

Figure Lengend Snippet: PMMA particles induce NF-κB activity and modulate extra-medullary hematopoiesis in the spleen. ( a ) PMMA particles induced NF-κB activation in immune cells. Total cells were isolated from the spleen from post PMMA intratibially injected animals. After removal of red blood cells, mononucleated cells were lysed to assess luciferase activity, which was normalized by protein concentration determined by standard BCA assay. ( b ) CD4 + T helper were further fractionated before measurements of luciferase activity normalized either by total protein input or cell number (not shown). ( c ) Decreased frequency of bone marrow CD4 + CD25 + Foxp3 + T REG in the spleen of PMMA injected animals. ( d ) Alterations of myeloid lineages. All columns in the graphs were represented as mean ± SD. *p < 0.05 by Student T-test.

Article Snippet: To transduce T REGS , 1 millions of freshly MACS-isolated naïve CD4+ T cells from mouse spleen were incubated overnight with 3 ml of retroviral stock supplemented with reagents from the T REG Cell Differentiation Kit (R&D Systems) and 10ug/ml of polybrene.

Techniques: Activity Assay, Activation Assay, Isolation, Injection, Luciferase, Protein Concentration, BIA-KA

Journal: Scientific Reports

Article Title: Inflammatory Responses Reprogram T REGS Through Impairment of Neuropilin-1

doi: 10.1038/s41598-019-46934-x

Figure Lengend Snippet: Increase of myeloid progenitors in response to PMMA is transient, while reduction of regulatory T cells is prolonged. ( a ) Frequency of bone marrow myeloid progenitor LSK, ( b ) CMP and ( c ) GMP 2, 4 and 7 days after intra-tibial injection of PMMA. ( d ) Frequency of T REG in the bone marrow (BM) and ( e ) in the spleen (Spl) over time. ( f ) Frequency of granulocytes in the mononuclear peripheral blood cells (PB/MNCs) over time. ( g ) IHC for Luciferase and FoxP3 from Control and PMMA-treated conditions. Arrows point to reactive staining ( h ) Quantification of normalized NF-κB reporter activity in WBM cells isolated from femur and tibia 7 days after intra-tibial (i.t.) injection of PMMA. # denotes femur adjacent to injected tibia. ( i ) Assessment of osteoclastogenic potential of WBM cells by ex vivo osteoclastogenesis assay (OCgenesis) 7 days after intra-tibial injection of PMMA. ( j ) % of CD4 + CD25 + FoxP3 lo RORγt + T cells in the bone marrow (BM) of mice treated as described in H-I. All columns in the graphs were represented as mean ± SD. *p < 0.05; **p < 0.005 by Student T-test.

Article Snippet: To transduce T REGS , 1 millions of freshly MACS-isolated naïve CD4+ T cells from mouse spleen were incubated overnight with 3 ml of retroviral stock supplemented with reagents from the T REG Cell Differentiation Kit (R&D Systems) and 10ug/ml of polybrene.

Techniques: Injection, Luciferase, Control, Staining, Activity Assay, Isolation, Ex Vivo

Journal: Scientific Reports

Article Title: Inflammatory Responses Reprogram T REGS Through Impairment of Neuropilin-1

doi: 10.1038/s41598-019-46934-x

Figure Lengend Snippet: Increase of T H 17 immunity, upregulation of inflammatory factors produced by effector T cells and recapitulation of the in vivo PMMA effect on T REG by ex vivo cultures. ( a – c ) CD4 + CD25 − effector T cells (T EFF ) were enriched by magnetic bead sorting (MACS) and lysed in Trizol reagent for RNA isolation. After cDNA synthesis through reverse transcription (RT), quantitative polymerase chain reaction (qPCR) was performed to assess mRNA expression of T H 17 markers including IL-17A, RORγt and RUNX1. ( d – f ) Expression of proinflammatory/pro-osteoclastogenic cytokines including M-CSF, RANKL and TNFα were also measured by qRT-PCR. Expression of tubulin was used as loading control. ΔCq values were calculated by Cq value of each given gene divided by that of tubulin. Plots were generated by values that were normalized with mock PBS injected control group. ( g – i ) Frequencies of CD4 + CD25 + Foxp3 + T REG in whole bone marrow (WBM), whole spleen (WSpl) and while lymph node (WLN) cells were assessed by flow cytometry after overnight incubation with 0.4% PMMA. ( j – l ) Frequencies CD4 + CD25 + CD44 + IL-17A + T EFF were also analyzed. All columns in the graphs were represented as mean ± SD. *p < 0.05; **p < 0.005 by Student T-test.

Article Snippet: To transduce T REGS , 1 millions of freshly MACS-isolated naïve CD4+ T cells from mouse spleen were incubated overnight with 3 ml of retroviral stock supplemented with reagents from the T REG Cell Differentiation Kit (R&D Systems) and 10ug/ml of polybrene.

Techniques: Produced, In Vivo, Ex Vivo, Isolation, cDNA Synthesis, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Quantitative RT-PCR, Control, Generated, Injection, Flow Cytometry, Incubation

Journal: Scientific Reports

Article Title: Inflammatory Responses Reprogram T REGS Through Impairment of Neuropilin-1

doi: 10.1038/s41598-019-46934-x

Figure Lengend Snippet: Inhibition of osteoclastogenesis by T REG in the presence of PMMA is Nrp1-mediated event. ( a ) MACS-purified T REG were either co-cultured with bone marrow derived macrophages (BMMs) or placed in the upper chambers of transwell cultures with BMMs in the lower chambers. RANKL and M-CSF were added at the beginning of cell culture and again after 2 days to promote osteoclastogenesis. PMMA particles were added to BMMs or T REGS as indicated. After 4 days, T REG cells were separated from cultures before TRAP staining was performed to visualize osteoclast formation. Results from BMM-T REG co-cultures and BMM-T REG transwell cultures are marked accordingly. ( b – e ) Isolated T REG from co-cultures and transwell upper chambers were lysed in Trizol reagent for RNA isolation. cDNAs were subsequently generated for qPCR analysis. ( f ) Immunostaining for Nrp1 in bone sections from control and PMMA-treated mice. ( g ) Osteoclast differentiation (+/−PMMA treated T REGS ) as described in panel A in the absence or presence of intact T REGS or T REGS transduced with scrambled (scr) shRNA or Nrp-1 shRNA. ( h ) Osteoclast differentiation (as described above) in the absence or presence of vehicle or PMMA-treated T REGS . Some conditions include overexpression of retroviral Nrp-1 in T REGS (as indicated). ( i ) Representative images of osteoclast cultures quantified in panel h. All columns in the graphs were represented as mean ± SD. *p < 0.05; **p < 0.005; ***p < 0.001 by Student T-test.

Article Snippet: To transduce T REGS , 1 millions of freshly MACS-isolated naïve CD4+ T cells from mouse spleen were incubated overnight with 3 ml of retroviral stock supplemented with reagents from the T REG Cell Differentiation Kit (R&D Systems) and 10ug/ml of polybrene.

Techniques: Inhibition, Purification, Cell Culture, Derivative Assay, Staining, Isolation, Generated, Immunostaining, Control, Transduction, shRNA, Over Expression, Retroviral