Journal: Immunity, Inflammation and Disease

Article Title: Distribution and chemotactic mechanism of CD4 + T cells in traumatic tracheal stenosis

doi: 10.1002/iid3.916

Figure Lengend Snippet: The expression levels of chemokines in BAL fluid and correlation analysis with chemokine receptors. (A) The statistical graph shows the concentration contrast of chemokines CCL1, CCL3, CCL4, CCL17, CCL22, CXCL9, CXCL10, and CXCL11 between the untreated and treated groups ( n = 10). (B) Correlation analysis between chemokines and paired chemokine receptors. Horizontal bars indicate means. Comparisons were made using paired t tests or Wilcoxon signed‐rank tests, and correlations were determined by Spearman's rank correlation coefficients. BAL, bronchoalveolar lavage.

Article Snippet: Grouping was as follows: PBS control group, BAL supernatant group, BAL + anti‐CCL1 (Thermo Fisher Scientific) group, BAL + anti‐CXCL10 (Thermo Fisher Scientific) group, and BAL + anti‐CCL22 (Thermo Fisher Scientific) group.

Techniques: Expressing, Concentration Assay

Journal: Immunity, Inflammation and Disease

Article Title: Distribution and chemotactic mechanism of CD4 + T cells in traumatic tracheal stenosis

doi: 10.1002/iid3.916

Figure Lengend Snippet: CCL1 and CCL22 were expressed in CD4 + T cells and CD68 + macrophages in TS BAL fluid. Macrophages and CD4 + T cells were tagged with anti‐CD68 and anti‐CD4 murine monoclonal antibodies, followed by rhodamine‐labeled goat antimouse serum staining (red). The expression levels of CCL1 and CCL22 were labeled with rabbit antihuman polyclonal antibodies and then labeled with FITC in goat antirabbit IgG (green). Two‐color immunofluorescence staining showed that some CD68 + macrophages and some CD4 + T cells expressed CCL1 and CCL22. Scale bar = 20 μm.

Article Snippet: Grouping was as follows: PBS control group, BAL supernatant group, BAL + anti‐CCL1 (Thermo Fisher Scientific) group, BAL + anti‐CXCL10 (Thermo Fisher Scientific) group, and BAL + anti‐CCL22 (Thermo Fisher Scientific) group.

Techniques: Labeling, Staining, Expressing, Immunofluorescence

Journal: Immunity, Inflammation and Disease

Article Title: Distribution and chemotactic mechanism of CD4 + T cells in traumatic tracheal stenosis

doi: 10.1002/iid3.916

Figure Lengend Snippet: CCL1 and CCL22 in BAL of TS showed chemotactic activity against CD4 + T cells. The Transwell method was adopted for migration assay. Purified CD4 + T cells (1 × 10 5 ) were plated into the upper chamber of a 6.5 mm Transwell (5 μm pore size). Three neutralizing antibodies (anti‐CCL1 0.1 μg/mL; anti‐CCL17 1 μg/mL; and anti‐CCL22 1 μg/mL) were added to the lower chamber. PBS was used as a negative control, and pure BAL was used as a positive control. After 16 h, CD4 + T cells on the upper side were removed, and cells in the lower chamber were fixed with methanol and stained with 0.1% crystal violet. (A) Representative microscopy pictures from six independent experiments, scale bar = 50 μm. (B) Chemotactic assay statistics analyzed by direct counting (number of migrated cells in 16 h) and indirect counting (optical density values). Data are expressed as the mean ± SEM. Between‐group comparisons were analyzed using one‐way ANOVA followed by Newman–Keuls tests. * p < .05, ** p < .01, *** p < .001, and **** p < .000.

Article Snippet: Grouping was as follows: PBS control group, BAL supernatant group, BAL + anti‐CCL1 (Thermo Fisher Scientific) group, BAL + anti‐CXCL10 (Thermo Fisher Scientific) group, and BAL + anti‐CCL22 (Thermo Fisher Scientific) group.

Techniques: Activity Assay, Migration, Purification, Negative Control, Positive Control, Staining, Microscopy, Chemotaxis Assay

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

Article Title: Human T-Lymphotropic Virus Type 1-Induced CC Chemokine Ligand 22 Maintains a High Frequency of Functional FoxP3 + Regulatory T Cells

doi: 10.4049/jimmunol.0903846

Figure Lengend Snippet: CCL22 concentration in plasma and culture supernatant. Concentration of CCL22 in fresh plasma (A) or in PBMC supernatant after 18 h incubation in vitro (B). Median values are shown under each box plot. The value is expressed in picograms per milliliter for plasma samples (A) or in picograms per milliliter per 106 PBMCs (B). The p values were calculated using Student t test.

Article Snippet: For CCL22 detection, after 12 h of incubation PBMCs were incubated for 4 h with Brefeldin A (Sigma-Aldrich) and stained with anti-human CCL22 (R&D Systems, Oxfordshire, U.K.).

Techniques: Concentration Assay, Clinical Proteomics, Incubation, In Vitro

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

Article Title: Human T-Lymphotropic Virus Type 1-Induced CC Chemokine Ligand 22 Maintains a High Frequency of Functional FoxP3 + Regulatory T Cells

doi: 10.4049/jimmunol.0903846

Figure Lengend Snippet: Tax expression and CCL22 expression. A, Expression of CCL22 and HTLV-1 Tax protein (left panel) and CCL22 and FoxP3 (right panel) in CD4+ cells from two independent representative HTLV-1–infected patients. B, Frequency in PBMCs of T cells of the following phenotypes: CCL22−CD4+Tax+, CCL22+CD4+Tax+, and CD4+CCL22+. The last column shows the frequency of CCL22+ cells in the CD4+Tax+ population.

Article Snippet: For CCL22 detection, after 12 h of incubation PBMCs were incubated for 4 h with Brefeldin A (Sigma-Aldrich) and stained with anti-human CCL22 (R&D Systems, Oxfordshire, U.K.).

Techniques: Expressing, Infection

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

Article Title: Human T-Lymphotropic Virus Type 1-Induced CC Chemokine Ligand 22 Maintains a High Frequency of Functional FoxP3 + Regulatory T Cells

doi: 10.4049/jimmunol.0903846

Figure Lengend Snippet: Modulation and induction of CCL22 expression. A, Concentration of CCL22 (picograms per milliliter per 106 PBMCs) in PBMC supernatant after 18 h incubation in vitro of whole PBMCs (left) and PBMCs depleted of CD8+ cells (right). Each line represents the rise in CCL22 concentration for each respective subject after CD8+ cell depletion. B, Concentration of CCL22 (picograms per milliliter per 106 PBMCs) detected in supernatant of Jurkat cells in the presence or absence of anti-CD3 coated beads and in the presence or absence of MT2 cells. C, Concentration of CCL22 (picograms per milliliter per 106 PBMCs) in supernatant of Jurkat cells transfected with different plasmids (5 μg/ml/106 cells). Supernatant was obtained 72 h after transfection. D, Correlation between the plasma concentration of CCL22 (picograms per milliliter) and the frequency of Tax expression in CD4+ T cells. The solid line represents the regression curve for patients with HAM/TSP and the dashed line represents the regression curve for ACs.

Article Snippet: For CCL22 detection, after 12 h of incubation PBMCs were incubated for 4 h with Brefeldin A (Sigma-Aldrich) and stained with anti-human CCL22 (R&D Systems, Oxfordshire, U.K.).

Techniques: Expressing, Concentration Assay, Incubation, In Vitro, Transfection, Clinical Proteomics

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

Article Title: Human T-Lymphotropic Virus Type 1-Induced CC Chemokine Ligand 22 Maintains a High Frequency of Functional FoxP3 + Regulatory T Cells

doi: 10.4049/jimmunol.0903846

Figure Lengend Snippet: Proviral load is associated with CCL22 concentration in plasma. There was a significant positive correlation between the proviral load and the plasma concentration of CCL22 in patients with HAM/TSP (n = 10) and ACs (n = 8).

Article Snippet: For CCL22 detection, after 12 h of incubation PBMCs were incubated for 4 h with Brefeldin A (Sigma-Aldrich) and stained with anti-human CCL22 (R&D Systems, Oxfordshire, U.K.).

Techniques: Concentration Assay, Clinical Proteomics

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

Article Title: Human T-Lymphotropic Virus Type 1-Induced CC Chemokine Ligand 22 Maintains a High Frequency of Functional FoxP3 + Regulatory T Cells

doi: 10.4049/jimmunol.0903846

Figure Lengend Snippet: CD4+FoxP3+ frequency in nonleukemic HTLV-1–infected individuals correlates with CCL22 concentration in plasma and supernatant of PBMCs. Correlation between the frequency of circulating CD4+FoxP3+ cells and the concentration of CCL22 in plasma from uninfected controls, ACs, and patients with HAM/TSP (A) and in patients with acute and chronic ATLL (B). The p values were determined by a two-tailed Spearman test.

Article Snippet: For CCL22 detection, after 12 h of incubation PBMCs were incubated for 4 h with Brefeldin A (Sigma-Aldrich) and stained with anti-human CCL22 (R&D Systems, Oxfordshire, U.K.).

Techniques: Infection, Concentration Assay, Clinical Proteomics, Two Tailed Test

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

Article Title: Human T-Lymphotropic Virus Type 1-Induced CC Chemokine Ligand 22 Maintains a High Frequency of Functional FoxP3 + Regulatory T Cells

doi: 10.4049/jimmunol.0903846

Figure Lengend Snippet: Functional effects of CCL22 on CD4+FoxP3+ cells of HTLV-1–infected patients. A, Rate of migration of CD4+FoxP3+ cells in a transwell assay. The results are expressed as the fold increase in the number of cells that migrated in response to different concentrations of CCL22, compared with medium alone, during 4 h incubation. The results represent the means of four independent experiments. B, Viability of each respective cell subset following 24h of incubation, according to the level of CCL22 chemokine added. The values represent the absolute number of viable cells, normalized to the control sample. The histogram represents the mean of four independent experiments.

Article Snippet: For CCL22 detection, after 12 h of incubation PBMCs were incubated for 4 h with Brefeldin A (Sigma-Aldrich) and stained with anti-human CCL22 (R&D Systems, Oxfordshire, U.K.).

Techniques: Functional Assay, Infection, Migration, Transwell Assay, Incubation, Control

Journal:

Article Title: Selective Induction of Th2-Attracting Chemokines CCL17 and CCL22 in Human B Cells by Latent Membrane Protein 1 of Epstein-Barr Virus

doi: 10.1128/JVI.78.4.1665-1674.2004

Figure Lengend Snippet: Secretion of chemokines by EBV-immortalized B cells. Freshly isolated CD19+ peripheral-blood B cells from three donors, three EBV-immortalized B-cell lines (BCLs), and two BL cell lines (Daudi and Raji) were seeded in six-well plates at 106/well and cultured for 3 days. TARC/CCL17, MDC/CCL22, I-309/CCL1, MEC/CCL28, MIP-1α/CCL3, MIP-1β/CCL4, and RANTES/CCL5 contents in the culture supernatants were measured by ELISA. For details, see Materials and Methods. All assays were done in triplicate. Each bar represents the mean plus the standard error of the mean from three separate experiments.

Article Snippet: Test samples were preincubated with or without anti-TARC/CCL17 monoclonal antibody (54026.11) (R&D Systems), anti-MDC/CCL22 monoclonal antibody (57226.11) (R&D Systems), polyclonal anti-MIP-1α/CCL3 (R&D Systems), polyclonal anti-MIP-1β/CCL4 (R&D Systems), or polyclonal anti-RANTES/CCL5 (R&D Systems) for 30 min at 0°C and then applied to the lower wells in a volume of 30 μl.

Techniques: Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Selective Induction of Th2-Attracting Chemokines CCL17 and CCL22 in Human B Cells by Latent Membrane Protein 1 of Epstein-Barr Virus

doi: 10.1128/JVI.78.4.1665-1674.2004

Figure Lengend Snippet: Role of EBV infection in the expression of chemokine genes in CD19+ B cells. Total RNA was prepared from peripheral-blood CD19+ B cells; CD19+ B cells infected with EBV for 2 (2d), 4, and 8 days; and EBV-immortalized B cells (BCL-NU). (A) Expression time course of EBV latent genes. RT-PCR analysis was carried out for EBNA2, LMP1, and GAPDH. For details, see Materials and Methods. Representative results from three separate experiments are shown. (B) Expression time course of chemokine genes. Real-time RT-PCR was carried out for TARC/CCL17, MDC/CCL22, I-309/CCL1, MEC/CCL28, MIP-1α/CCL3, MIP-1β/CCL4, and RANTES/CCL5. For details, see Materials and Methods. Data are shown as the mean plus the standard error of the mean from three separate experiments.

Article Snippet: Test samples were preincubated with or without anti-TARC/CCL17 monoclonal antibody (54026.11) (R&D Systems), anti-MDC/CCL22 monoclonal antibody (57226.11) (R&D Systems), polyclonal anti-MIP-1α/CCL3 (R&D Systems), polyclonal anti-MIP-1β/CCL4 (R&D Systems), or polyclonal anti-RANTES/CCL5 (R&D Systems) for 30 min at 0°C and then applied to the lower wells in a volume of 30 μl.

Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal:

Article Title: Selective Induction of Th2-Attracting Chemokines CCL17 and CCL22 in Human B Cells by Latent Membrane Protein 1 of Epstein-Barr Virus

doi: 10.1128/JVI.78.4.1665-1674.2004

Figure Lengend Snippet: Roles of EBNA2 and LMP1 in the expression of chemokine genes. Total RNA was prepared from EBV-immortalized B-cell lines (BCL-NU and BCL-SH), BJAB, BJAB infected with EBV (BJAB-EBV), BJAB stably transfected with vector only (BJAB-vector; three clones), BJAB stably transfected with EBNA2 (BJAB-EBNA2; three clones), and BJAB stably transfected with LMP1 (BJAB-LMP1; three clones). RT-PCR analysis was carried out for TARC/CCL17, MDC/CCL22, I-309/CCL1, MEC/CCL28, MIP-1α/CCL3, MIP-1β/CCL4, RANTES/CCL5, EBNA2, LMP1, and GAPDH. For details, see Materials and Methods. Representative results from three independent experiments are shown.

Article Snippet: Test samples were preincubated with or without anti-TARC/CCL17 monoclonal antibody (54026.11) (R&D Systems), anti-MDC/CCL22 monoclonal antibody (57226.11) (R&D Systems), polyclonal anti-MIP-1α/CCL3 (R&D Systems), polyclonal anti-MIP-1β/CCL4 (R&D Systems), or polyclonal anti-RANTES/CCL5 (R&D Systems) for 30 min at 0°C and then applied to the lower wells in a volume of 30 μl.

Techniques: Expressing, Infection, Stable Transfection, Transfection, Plasmid Preparation, Clone Assay, Reverse Transcription Polymerase Chain Reaction

Journal:

Article Title: Selective Induction of Th2-Attracting Chemokines CCL17 and CCL22 in Human B Cells by Latent Membrane Protein 1 of Epstein-Barr Virus

doi: 10.1128/JVI.78.4.1665-1674.2004

Figure Lengend Snippet: Effects of various signaling inhibitors on the expression of chemokine genes in EBV-immortalized B cells and BJAB-LMP1. EBV-immortalized B cells (A) or BJAB-LMP1 cells (B) were treated for 12 h with or without PD98059 (the inhibitor of the MEK/ERK pathway) at 30 μM, SB202190 (the inhibitor of the p38/ATF2 pathway) at 10 μM, JNK inhibitor (the inhibitor of the JNK/AP-1 pathway) at 30 μM, JAK3 inhibitor (the inhibitor of the JAK3/STAT pathway) at 30 μM, AG490 (the inhibitor of the JAK2/STAT pathway) at 20 μM, and BAY11-7082 (the inhibitor of the TRAF/NF-κB pathway) at 5 μM. The concentrations of the inhibitors used were carefully chosen as optimal based on preliminary experiments. Total RNA was prepared, and quantitative real-time RT-PCR was carried out for TARC/CCL17, MDC/CCL22, I-309/CCL1, MEC/CCL28, MIP-1α/CCL3, MIP-1β/CCL4, and RANTES/CCL5. For details, see Materials and Methods. The data are shown as means plus standard errors of the mean from three separate experiments.

Article Snippet: Test samples were preincubated with or without anti-TARC/CCL17 monoclonal antibody (54026.11) (R&D Systems), anti-MDC/CCL22 monoclonal antibody (57226.11) (R&D Systems), polyclonal anti-MIP-1α/CCL3 (R&D Systems), polyclonal anti-MIP-1β/CCL4 (R&D Systems), or polyclonal anti-RANTES/CCL5 (R&D Systems) for 30 min at 0°C and then applied to the lower wells in a volume of 30 μl.

Techniques: Expressing, Quantitative RT-PCR

Journal:

Article Title: Selective Induction of Th2-Attracting Chemokines CCL17 and CCL22 in Human B Cells by Latent Membrane Protein 1 of Epstein-Barr Virus

doi: 10.1128/JVI.78.4.1665-1674.2004

Figure Lengend Snippet: Mechanism of transcriptional activation of MDC/CCL22 promoter by LMP1. (A) Schematic depiction of the potential transcriptional elements in the promoter regions of TARC/CCL17, MDC/CCL22, I-309/CCL1, MEC/CCL28, MIP-1α/CCL3, MIP-1β/CCL4, and RANTES/CCL5. The TFSEARCH program was used to localize potential regulatory elements. (B) Deletion analysis. BJAB-LMP1 and BJAB-vector were transfected with a series of the MDC/CCL22 promoter-luciferase constructs: pGL3, pGL3-MDC/CCL22(−722/−11), pGL3-MDC/CCL22(−553/−11), pGL3-MDC/CCL22(−476/−11), pGL3-MDC/CCL22(−289/−11), pGL3-MDC/CCL22(−114/−11), and pGL3-MDC/CCL22(−80/−11). (C) Mutation analysis. BJAB-LMP1 and BJAB-vector were transfected with the MDC/CCL22 promoter-luciferase constructs pGL3, pGL3-MDC/CCL22(−722/−11), pGL3-MDC/CCL22-ΔSTAT, pGL3-MDC/CCL22-ΔNF-κB (1), pGL3-MDC/CCL22-ΔNF-κB (2), pGL3-MDC/CCL22-ΔNF-κB (1) and (2), pGL3-MDC/CCL22-ΔAP-1, and pGL3-MDC/CCL22-ΔNF-κB (1) and (2) +AP-1. After 27 to 36 h, relative luciferase activity was determined with a luminometer. Promoter activation by LMP1 was expressed as the induction (n-fold) of luciferase activity in BJAB-LMP1 versus that in BJAB-vector. Each bar represents the mean plus standard error of the mean from three separate experiments. (D) Transactivation of MDC/CCL22 promoter by overexpression of NF-κB p65. BJAB cells were cotransfected with MDC/CCL22 promoter-luciferase constructs, namely, pGL3, pGL3-MDC/CCL22(−722/−11), pGL3-MDC/CCL22-ΔNF-κB (1), pGL3-MDC/CCL22-ΔNF-κB (2), and pGL3-MDC/CCL22-ΔNF-κB (1) and (2), and an expression vector for (His)6-p65 or a control vector. After 27 to 36 h, relative luciferase activity was determined with a luminometer. Promoter activation by p65 was expressed as the induction (n-fold) of luciferase activity in BJAB transfected with p65-vector versus that in BJAB transfected with control vector. Each bar represents the mean plus standard error of the mean from three separate experiments.

Article Snippet: Test samples were preincubated with or without anti-TARC/CCL17 monoclonal antibody (54026.11) (R&D Systems), anti-MDC/CCL22 monoclonal antibody (57226.11) (R&D Systems), polyclonal anti-MIP-1α/CCL3 (R&D Systems), polyclonal anti-MIP-1β/CCL4 (R&D Systems), or polyclonal anti-RANTES/CCL5 (R&D Systems) for 30 min at 0°C and then applied to the lower wells in a volume of 30 μl.

Techniques: Activation Assay, Plasmid Preparation, Transfection, Luciferase, Construct, Mutagenesis, Activity Assay, Over Expression, Expressing

Journal:

Article Title: Selective Induction of Th2-Attracting Chemokines CCL17 and CCL22 in Human B Cells by Latent Membrane Protein 1 of Epstein-Barr Virus

doi: 10.1128/JVI.78.4.1665-1674.2004

Figure Lengend Snippet: Elevated serum MDC/CCL22 and MEC/CCL28 contents in IM. Serum samples were obtained from healthy donors (n = 9) and patients with IM (n = 9). Serum TARC/CCL17, MDC/CCL22, I-309/CCL1, MEC/CCL28, MIP-1α/CCL3, MIP-1β/CCL4, and RANTES/CCL5 contents were measured by ELISA. All assays were done in duplicate. The horizontal lines indicate mean values. Statistical significance was determined by the unpaired Welch's t test. *, P < 0.05; **, P < 0.01.

Article Snippet: Test samples were preincubated with or without anti-TARC/CCL17 monoclonal antibody (54026.11) (R&D Systems), anti-MDC/CCL22 monoclonal antibody (57226.11) (R&D Systems), polyclonal anti-MIP-1α/CCL3 (R&D Systems), polyclonal anti-MIP-1β/CCL4 (R&D Systems), or polyclonal anti-RANTES/CCL5 (R&D Systems) for 30 min at 0°C and then applied to the lower wells in a volume of 30 μl.

Techniques: Enzyme-linked Immunosorbent Assay