Journal: medRxiv

Article Title: Trajectories of microbiome-derived bile acids in early life – insights into the progression to islet autoimmunity

doi: 10.1101/2025.02.18.25322275

Figure Lengend Snippet: a. Schematic of the Th17 and iTreg differentiation protocol for primary human naïve CD4 + CD25 - T cells isolated from the umbilical cord blood of healthy neonates. CD4 + CD25 - T cells were activated with anti-CD3/anti-CD28 and differentiated into Th17 or iTreg cells in the presence of corresponding cytokines for 3 days. DMSO control or conjugated bile acids (Asn-UDCA, Ser-CDCA, Tyr-CDCA, and unconjugated UDCA) at 100 µM were added on day 0 of differentiation. b-e. IL-17a secretion in the supernatant of Th17 cultures treated with Asn-UDCA ( b ), Ser-CDCA ( c ), Tyr-CDCA ( d ), and UDCA ( e ) was quantified on day 3 of differentiation from four biological replicates using ELISA. f-i. Intracellular Foxp3 protein expression in iTregs cultured with Asn-UDCA ( f ), Ser-CDCA ( g ), Tyr-CDCA ( h ), or UDCA ( i ) was assessed on day 3 of differentiation by flow cytometry. Geometric mean fluorescence intensity (MFI) values are shown for four biological replicates. Statistical significance was determined using paired, two-tailed Student’s t-test.

Article Snippet: After differentiation, secreted IL-17a levels were determined from Th17 cell-culture supernatants at 72 hours using the human IL-17a DuoSet ELISA kit (R&D Systems, Cat# DY317-05, DY008).

Techniques: Isolation, Control, Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, Flow Cytometry, Fluorescence, Two Tailed Test

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: Immunity

Article Title: CD95-Mediated Calcium Signaling Promotes T Helper 17 Trafficking to Inflamed Organs in Lupus-Prone Mice

doi: 10.1016/j.immuni.2016.06.028

Figure Lengend Snippet: Serum CD95L in SLE Patients Induces Transmigration of T Lymphocytes (A) Soluble CD95L levels were measured in serum from newly diagnosed SLE patients (n = 34) and healthy donors (n = 8) via ELISA (Student’s t test). (B) CD95L in SLE serum was fractionated by size-exclusion column chromatography and measured via ELISA. Inset : CD95L was immunoprecipitated from gel-filtration fractions 40–46 and 76–78 and subjected to immunoblotting. The image is representative of gel-filtration analysis of four different patients. (C) CD95L and IL-17 staining in inflamed skin samples from lupus patients or healthy mastectomy subjects. Numbers represent different patients. The scale bar represents 100 μm. (D) CD95L, CD31, CD4, and IL-17 staining in inflamed skin samples from an SLE patient. “V” represents an endothelial vessel, and arrowheads identify marker-expressing cells. (E) Densitometric analysis of CD95L and IL-17 staining in different patients. (F) Transmigration of human T cell subpopulations in the presence of serum from SLE patients or healthy donors. (G) Th17 T cell transmigration in the presence of SLE serum containing the indicated concentrations of CD95-Fc. Undifferentiated Th 0 T cells served as controls. Data represent means ± SD of five individual serum donors. (H) CD4 + T cell transmigration with or without cl-CD95L (100 ng/mL). Data represent means ± SD of three independent experiments. (I) Treg and Th17 cell transmigration with or without cl-CD95L (100 ng/mL) Data represent means ± SD of three independent experiments (two-way ANOVA).

Article Snippet: PBMCs were then subjected to selection with a cocktail of antibody-coated magnetic beads: CCR6 + CXCR3 − CD4 + cells (Th17 cells) were sorted with Human Th17 Enrichment kit (STEMCELL Technologies), and CD4 + CD25 + CD127 − Treg cells were isolated with MACS column (Miltenyi Biotec).

Techniques: Transmigration Assay, Enzyme-linked Immunosorbent Assay, Column Chromatography, Immunoprecipitation, Filtration, Western Blot, Staining, Marker, Expressing

Journal: Immunity

Article Title: CD95-Mediated Calcium Signaling Promotes T Helper 17 Trafficking to Inflamed Organs in Lupus-Prone Mice

doi: 10.1016/j.immuni.2016.06.028

Figure Lengend Snippet: Transcriptomic Signature in Human Th17 and Treg Cells Stimulated with cl-CD95L (A) Venn diagram comparing the genes differentially expressed between untreated and cl-CD95L-treated T cell subsets. (B) Heat map depicting the relative n-fold change in the amount of transcripts significantly (p ≤ 0.05) and differentially expressed between Th17 and Treg cells stimulated with cl-CD95L (100 ng/mL). Data for each experimental group (n = 2 per condition) are shown. The color gradient indicates n-fold change, as shown. (C) Pathway enrichment analysis of genes whose expression is significantly modulated by cl-CD95L in Th17 and Treg cells and associated p values. (D) Left panel : endothelial transmigration of human Th17 and Treg cells was evaluated in the presence or absence of cl-CD95L (100 ng/mL) in the Boyden chamber. Right panel : Th17 cells were pre-treated with FTY720 (1 μM), TY-52156 (10 μM), VPC-23019 (10 μM), W146 (1 μM), or CAY-1044 (1 μM) and then exposed to cl-CD95L (100 ng/mL), and endothelial transmigration was evaluated via the Boyden chamber. Data represent means ± SD of three independent experiments.

Article Snippet: PBMCs were then subjected to selection with a cocktail of antibody-coated magnetic beads: CCR6 + CXCR3 − CD4 + cells (Th17 cells) were sorted with Human Th17 Enrichment kit (STEMCELL Technologies), and CD4 + CD25 + CD127 − Treg cells were isolated with MACS column (Miltenyi Biotec).

Techniques: Expressing, Transmigration Assay

Journal: Immunity

Article Title: CD95-Mediated Calcium Signaling Promotes T Helper 17 Trafficking to Inflamed Organs in Lupus-Prone Mice

doi: 10.1016/j.immuni.2016.06.028

Figure Lengend Snippet: Cl-CD95L Is a Chemoattractant for Th17 Cells In Vivo Mice received a single injection of cl-CD95L (200 ng/animal) or vehicle and were sequentially sampled. (A) T cell populations were obtained from tissues or peritoneal cavity washes. Cells were then restimulated in the presence of PMA and ionomycin for 4 hr and then analyzed by flow cytometry. Cells were identified as follows; Th1 (CD4 + IFN-γ + ), Th17 (CD4 + IL-17 + ), and Treg (CD4 + Foxp3 + ). Numbers of infiltrating cells were calculated. (B) Ratios of Th17/Th1 cells per organ were determined 24 hr after injection. Data represent two independent experiments with six mice/group; means ± SEM are displayed.

Article Snippet: PBMCs were then subjected to selection with a cocktail of antibody-coated magnetic beads: CCR6 + CXCR3 − CD4 + cells (Th17 cells) were sorted with Human Th17 Enrichment kit (STEMCELL Technologies), and CD4 + CD25 + CD127 − Treg cells were isolated with MACS column (Miltenyi Biotec).

Techniques: In Vivo, Injection, Flow Cytometry

Journal: Immunity

Article Title: CD95-Mediated Calcium Signaling Promotes T Helper 17 Trafficking to Inflamed Organs in Lupus-Prone Mice

doi: 10.1016/j.immuni.2016.06.028

Figure Lengend Snippet: CD95 Induces a DD-Independent Ca 2+ Response (A) CEM T cells were stimulated with cl-CD95L (100 ng/mL). Cells were lysed, and CD95 was immunoprecipitated. The protein complex was resolved by SDS-PAGE and subjected to immunoblotting. Total lysates served as controls. The column marked “B” indicates treatment with beads alone. Data are representative of three independent experiments. (B) Left panel : Th17 cells from peripheral blood were stimulated with cl-CD95L (100 ng/mL) for indicated times. PLA was performed with anti-CD95 and anti-PLCγ1 mAbs. Nuclei were stained in blue (DAPI). Red dots were observed when the distance between anti-CD95 and anti-PLCγ1 mAbs was close (≈16 nm). Right panels : Red dots were counted in 200 cells taken from different fields. Data represent means ± SD of three independent experiments. (C) Schematic diagram of CD95 constructs. (D) CEM-IRC cells expressing GFP alone or the GFP-fused CD95 constructs shown in (C) were loaded with the Ca 2+ probe, Fluo2-AM (1 μM). Cells were stimulated with cl-CD95L (100 ng/mL; arrow), and the intracellular calcium concentration ([Ca 2+ ] i ) was monitored. Data are given as means ± SD of three experiments performed independently on n = 20 cells. (E) HEK cells transfected with the indicated constructs were stimulated with CD95L (100 ng/mL) for indicated times. The CD95 protein complex was immunoprecipitated from cell lysates and subjected to immunoblotting, as indicated. Total lysates served as controls. The column marked “B” indicates treatment with beads alone. Data are representative of three independent experiments. (F) Activated PBLs were pre-incubated for 1 hr with TAT-control or TAT-CID (10 μM) and stimulated with cl-CD95L (100 ng/mL) for the indicated times. The CD95 protein complex was immunoprecipitated from cell lysates and subjected to immunoblotting, as indicated. Total lysates served as controls. The column marked “B” indicates a treatment with beads alone. Data are representative of three independent experiments. (G) Activated PBLs from healthy donors were loaded with FuraPE3-AM (1 μM) and pre-treated for 1 hr with TAT-control or TAT-CID (10 μM). Cells were stimulated with cl-CD95L (100 ng/mL; arrow). Data represent means ± SD.

Article Snippet: PBMCs were then subjected to selection with a cocktail of antibody-coated magnetic beads: CCR6 + CXCR3 − CD4 + cells (Th17 cells) were sorted with Human Th17 Enrichment kit (STEMCELL Technologies), and CD4 + CD25 + CD127 − Treg cells were isolated with MACS column (Miltenyi Biotec).

Techniques: Immunoprecipitation, SDS Page, Western Blot, Staining, Construct, Expressing, Concentration Assay, Transfection, Incubation, Control

Journal: Immunity

Article Title: CD95-Mediated Calcium Signaling Promotes T Helper 17 Trafficking to Inflamed Organs in Lupus-Prone Mice

doi: 10.1016/j.immuni.2016.06.028

Figure Lengend Snippet: TAT-CID Alters Immunological Parameters in Lupus-Prone Mice (A) Transmigration of mouse Th17 cells was monitored with the indicated concentrations of TAT-CID. (B–D) C57BL/6 mice were injected with TAT-control or TAT-CID (40 mg/kg) 2 hr prior to intraperitoneal injection of cl-CD95L (200 ng) or vehicle. Animals were examined 24 hr after cl-CD95L injection. (B) Total cell counts in the peritoneal cavity are shown. (C) PECs were subjected to magnetic bead separation to identify the percentage of infiltrating CD4 + CD62L − (activated) T cells. (D) IL-17A concentrations in the peritoneal cavity were measured via ELISA (two-way ANOVA). Data in (B)–(D) represent two independent experiments performed with six mice/group. Data are means ± SEM. (E–I) MRL. Fas lpr/+ mice received either TAT-CID or TAT-control for 5 weeks. (E) Upon completion of the experimental protocol, ratios of spleen weight to body weight of individual animals were measured and compared to those of age-matched MRL and homozygous MRL. Fas lpr/lpr mice. (F) Total cell number in the spleen is shown. (G) Cellular composition of the spleen was determined in regard to the number of CD4 + CD62L − T cells. (H) mRNA expression levels of il-23r, ccr6 , and ror-γt in cells from (C). (I) Isolated T cells from (G) were re-stimulated with anti-CD3 mAb for 72 hr. IL-17A was then quantified by ELISA (unpaired Student’s t test).

Article Snippet: PBMCs were then subjected to selection with a cocktail of antibody-coated magnetic beads: CCR6 + CXCR3 − CD4 + cells (Th17 cells) were sorted with Human Th17 Enrichment kit (STEMCELL Technologies), and CD4 + CD25 + CD127 − Treg cells were isolated with MACS column (Miltenyi Biotec).

Techniques: Transmigration Assay, Injection, Control, Enzyme-linked Immunosorbent Assay, Expressing, Isolation

Journal: The Journal of investigative dermatology

Article Title: IL-17 and TNF synergistically modulate cytokine expression while suppressing melanogenesis: potential relevance to psoriasis

doi: 10.1038/jid.2013.237

Figure Lengend Snippet: (a) Cytokines and growth factors inductions in melanocytes treated with IL-17 and/or TNF for 24 h. (b-c) Intracellular cytokine staining shows induction of CXCL1 and IL-8 in melanocytes after treatment with TNF and IL-17 for 24 h. Median fluorescent intensity is indicated next to each condition. (d) CXCL1 secretion was assessed by ELISA after 24 h in culture. (e) IL-8 secretion was assessed by ECL assay after 24 h in culture. (*P<0.05, **P<0.01, vs. Ctrl). Data represent results from three independent cultures using melanocytes from three different skin donors.

Article Snippet: Kotobuki T. et al reported that a combination of 4 cytokines (i.e. TNF, IL-1β, IL-6 and IL-17) could inhibit melanin production ( Kotobuki et al. , 2012 ).

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: IL-17 and TNF synergistically modulate cytokine expression while suppressing melanogenesis: potential relevance to psoriasis

doi: 10.1038/jid.2013.237

Figure Lengend Snippet: (a) Decreased expressions of pigmentation genes (paired lesional vs. non-lesional skin) in a meta-analysis derived of transcriptome of over 190 psoriasis patients (P<0.05, FDR<0.05) (b) qRT-PCR analysis confirms suppression of pigmentation genes in paired psoriasis lesional vs. non-lesional skin (n=6). (*P<0.05, *P<0.01). Gene expression changes for each patient were represented by a line with a different color. (c) Increased expression of β-defensin 3 in lesional psoriasis skin, compared to non-lesional skin (n=10). Bar=100μm (d) IL-17 and TNF induces the expression of β-defensin 3 , an antagonist for melanocortin-1 receptor, in keratinocytes after 24 h treatment with individual cytokines: IL-17 (200ng/mL), IL-22 (200ng/mL), IFNγ (20ng/mL), and TNF (10ng/mL) (*P < 0.05; **P < 0.01; ***P<0.001, vs. Ctrl).

Article Snippet: Kotobuki T. et al reported that a combination of 4 cytokines (i.e. TNF, IL-1β, IL-6 and IL-17) could inhibit melanin production ( Kotobuki et al. , 2012 ).

Techniques: Derivative Assay, Quantitative RT-PCR, Gene Expression, Expressing

Journal: The Journal of investigative dermatology

Article Title: IL-17 and TNF synergistically modulate cytokine expression while suppressing melanogenesis: potential relevance to psoriasis

doi: 10.1038/jid.2013.237

Figure Lengend Snippet: (a) IL-17 secreted by skin Th17 cells, together with TNF secreted by T cells, dendritic cells, can jointly induce mitogenic cytokines in both keratinocytes (KC) and melanocytes (MC), creating a milieu that supports cell proliferation. This leads to an increase in melanocytes numbers at psoriasis lesions. This is accompanied by a synergistic suppression of pigmentation signaling and melanin synthesis. (b) Therapeutic neutralization by monoclonal antibodies can lead to a rapid restoration of pigment function within 2-4 weeks. An elevated number of melanocytes in lesional skin will be producing abundant melanin, which persists in keratinocytes during early phases of clinical improvement, leading to post-inflammatory hyper-pigmentation.

Article Snippet: Kotobuki T. et al reported that a combination of 4 cytokines (i.e. TNF, IL-1β, IL-6 and IL-17) could inhibit melanin production ( Kotobuki et al. , 2012 ).

Techniques: Neutralization, Bioprocessing

Journal: Journal of Immunology Research

Article Title: New Insights about Treg and Th17 Cells in HIV Infection and Disease Progression

doi: 10.1155/2015/647916

Figure Lengend Snippet: Immunophenotyping of Treg and Th17 cells and their precursors in different studies.

Article Snippet: Memory Th17 , CD45RA − CCR6 + CCR4 + CXCR3 − CD4 + , Gosselin et al. [ ]; Becattini et al. [ ]; Acosta-Rodriguez et al. [ ] .

Techniques:

Journal: Journal of Immunology Research

Article Title: New Insights about Treg and Th17 Cells in HIV Infection and Disease Progression

doi: 10.1155/2015/647916

Figure Lengend Snippet: The interaction network between transcriptional factors, cytokines, chemokines, and their receptors in Th17 and Treg cells. The fine-tuning of Th17/Treg balance is regulated by expression of transcription factors that are activated by cytokines milieu and their receptors. TGF- β along with mainly IL-6 induces RORc, ROR- α , or STAT3 expression to differentiate Th17 cells while that in combination with IL-2 induces FoxP3 expression to differentiate Treg cells, while homing and immunological cells recruitment of both cell subsets are powerful mechanism mediated by chemokines and their chemokine receptors such as CCR6, CCR4, or CXCR3 which facilitates the recruitment of suppressive Treg and inflammatory effector Th17 cells (e.g., by means of CCR6-CCL20) into the site infection or injured tissue. Of note, other immunological cells, as dendritic cells, influence this balance because they produce cytokines, chemokines, and other molecules that participate in this interaction network.

Article Snippet: Memory Th17 , CD45RA − CCR6 + CCR4 + CXCR3 − CD4 + , Gosselin et al. [ ]; Becattini et al. [ ]; Acosta-Rodriguez et al. [ ] .

Techniques: Expressing, Infection

Journal: Molecular Medicine

Article Title: Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes

doi: 10.1186/s10020-024-00791-1

Figure Lengend Snippet: Fluvoxamine decreased the population of Th1 and Th17 cells in the PLNs, spleen, pancreas and peripheral blood. A–C PLN and splenic cells from 12-week-old fluvoxamine-treated and vehicle-treated mice were harvested and subject to flow cytometry analysis. Frequencies of A CD4 + IFN-γ + (Th1), B CD4 + IL-17A + (Th17), and C CD4 + CD25 + Foxp3 + (Treg) subsets were examined. D–F Blood from the mouse tail vein were collected and subjected to flow cytometry analysis. Frequencies of D Th1, E Th17, and F Treg subsets are shown as representative dot plot graphs. G, H Flow cytometry analysis of pancreatic cells from 12-week-old fluvoxamine-treated and vehicle-treated mice. Representative flow cytometry plots and frequencies of G Th1 and H Th17 effector T cells (n = 5 per group). I–K CD4 + T cells from peripheral blood of newly onset diabetic T1D subjects were stimulated with fluvoxamine or vehicle for 48 h, and the ratio of subtypes in CD4 + T cells were measured by flow cytometry. The cell frequency of I Th1 and J Th17 as well as K Treg cells was determined. The frequency of T cell subsets was investigated in ten donors. Data are expressed as mean ± SEM. Statistical significance was calculated by unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, not significant

Article Snippet: Th17 cells induced by IL-6, IL-23, and IL-1β are generally thought to be highly pathogenic, which tend to exhibit characteristics similar as Th1 cells in the context of T1D (Haskins and Cooke ).

Techniques: Flow Cytometry

Journal: Molecular Medicine

Article Title: Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes

doi: 10.1186/s10020-024-00791-1

Figure Lengend Snippet: Fluvoxamine suppresses the differentiation of Th1 and Th17 cells rather than disturbing their proliferation and apoptosis. Splenic CD4 + T cells were isolated and stimulated with fluvoxamine (10 μM) or vehicle. The percentage of proliferated CD4 + T cells was defined by A Carboxyfluorescein Succinimidylester (CFSE) assay after 3 days of culture, and B apoptosis of T cells was determined by PI and Annexin V staining. Representative histograms of CFSE in C Th1 and D Th17 cells. Representative FACS plots and frequencies of active caspase-3 in E Th1 and F Th17 cells. Each dot represents the mean of three biological replicates. PLN cells from 12-week-old fluvoxamine- and vehicle-treated mice were harvested and subject to flow cytometry analysis. Representative FACS plots and frequencies of G CD4 + Ki67 + T cells and H apoptotic CD4 + T cells determined by PI and Annexin V staining are shown. Percentage of Ki67 + proliferative I effector Th1 and J Th17 cells are shown by flow cytometry. K Apoptotic effector Th1 and L Th17 cells were indicated by staining active caspase-3 positive cells (n = 5 per group). Naïve CD4 + T cells purified from splenocytes were cultured under Th1 and Th17 conditions in vitro for 3 days in the presence of fluvoxamine or vehicle. M Th1 and N Th17 polarization efficiency was analyzed by flow cytometry. Each dot represents the mean of three biological replicates. Data are expressed as mean ± SEM. Statistical significance was calculated by unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, not significant

Article Snippet: Th17 cells induced by IL-6, IL-23, and IL-1β are generally thought to be highly pathogenic, which tend to exhibit characteristics similar as Th1 cells in the context of T1D (Haskins and Cooke ).

Techniques: Isolation, CFSE Assay, Staining, Flow Cytometry, Purification, Cell Culture, In Vitro

Journal: Molecular Medicine

Article Title: Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes

doi: 10.1186/s10020-024-00791-1

Figure Lengend Snippet: Fluvoxamine represses Th1 and Th17 differentiation and glycolysis by regulating the PI3K/AKT axis. A Genes downregulated in fluvoxamine-treated CD4 + T were subjected to KEGG pathway enrichment analysis. B Results for Gene Set Enrichment Analysis (GSEA) of PI3K-AKT signaling pathways. C, D The purified CD4 + T cells were cultured with fluvoxamine (0 μM, 5 μM, 10 μM) for 24 h. The protein expression of p-PI3K, p-AKT, PI3K and AKT was assessed. CD4 + T cells were cultured with fluvoxamine, vehicle or PI3K activator 740 Y-P for 24 h and E ECAR was analyzed by an extracellular flux analyzer. F Results for glycolysis and glycolytic capacity in CD4 + T cells with different treatment. G and H Naïve CD4 + T cells isolated from spleen were exposed to Th1-or Th17- inducing conditions under indicated culture conditions. G Th1 and H Th17 polarization efficiency were analyzed by flow cytometry. Each dot represents the mean of three biological replicates. Data are expressed as mean ± SEM. Significance was determined by one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001, and ns, not significant

Article Snippet: Th17 cells induced by IL-6, IL-23, and IL-1β are generally thought to be highly pathogenic, which tend to exhibit characteristics similar as Th1 cells in the context of T1D (Haskins and Cooke ).

Techniques: Protein-Protein interactions, Purification, Cell Culture, Expressing, Isolation, Flow Cytometry

Journal: Molecular Medicine

Article Title: Fluvoxamine inhibits Th1 and Th17 polarization and function by repressing glycolysis to attenuate autoimmune progression in type 1 diabetes

doi: 10.1186/s10020-024-00791-1

Figure Lengend Snippet: The scheme of fluvoxamine attenuates autoimmune progression in T1D setting. Fluvoxamine treatment inhibits glycolysis via restraining PI3K/AKT signaling. This metabolic change results in impaired differentiation and function of Th1 and Th17 cells. Consequently, administration of fluvoxamine delays diabetes onset and prevents autoimmune progression in T1D setting

Article Snippet: Th17 cells induced by IL-6, IL-23, and IL-1β are generally thought to be highly pathogenic, which tend to exhibit characteristics similar as Th1 cells in the context of T1D (Haskins and Cooke ).

Techniques: