Journal: Journal of experimental & clinical cancer research : CR

Article Title: High and selective cytotoxicity of ex vivo expanded allogeneic human natural killer cells from peripheral blood against bladder cancer: implications for natural killer cell instillation after transurethral resection of bladder tumor.

doi: 10.1186/s13046-024-02955-7

Figure Lengend Snippet: Fig. 7 The number of T cells migrating across the transwell and the protein expression levels of chemokines mediating T-cell recruitment. (A) The activat ed CD3+ T cells were seeded into transwells with simple media and conditioned media from T24 cell cultures, NK cell cultures, and NK and T24 cell cocul tures. Summary data of the cumulative numbers of CD3+ T cells that migrated across the transwell in each condition are shown (n = 3 per group). ns, not significant; *p < 0.05 ; (B) Protein expression levels of chemokines in the supernatant from T24 culture, NK culture, and NK + T24 coculture measured using ELISA (n = 3 per group). Data expressed as mean ± SD were plotted. Abbreviations: CCL: C-C-motif ligand; CXCL:C-X-C-motif ligand; XCL1:X-C-motif ligand

Article Snippet: For chemokine release assay in transwell experiment, concentrations of human chemokines in the supernatant including C-C-motif ligand 1 (CCL1), CCL2, CCL4, CCL20, C-XC-motif ligand 1 (CXCL1), CXCL2, CXCL3, CXCL8, CXCL16, and C-motif ligand 1 (XCL1) were measured using human ELISA kits from CUSABIO for CCL1 (#CSB-EL004774HU), CCL2 (#CSB-Eq.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: High and selective cytotoxicity of ex vivo expanded allogeneic human natural killer cells from peripheral blood against bladder cancer: implications for natural killer cell instillation after transurethral resection of bladder tumor.

doi: 10.1186/s13046-024-02955-7

Figure Lengend Snippet: Fig. 8 Receptors of the chemokines expressed on CD3+ T cells and T cell count migrating across the transwell after blocking CCL1/2/20 in the superna tant of coculture medium. (A) The expressions of the chemokine receptors on CD3+CD56− T cells were detected using flow cytometry (n = 3 for all mol ecules). The representative images are shown. (B) Summary data of the cumulative numbers of T cells determined using a fluorescence cell analyzer after being harvested from the lower chambers, which were added into blocking antibodies against CCL1 (0.25 µg/mL), CCL2 (40 ng/mL), or CCL20 (10 µg/ mL) (n = 3 for each). Data are shown as mean ± SD. Statistical significance was determined using an unpaired t-test. *p < 0.05. Abbreviations: CXCR: C-X-C chemokine receptor; CCR:C-C chemokine receptor; XCR:X-C chemokine receptor

Article Snippet: For chemokine release assay in transwell experiment, concentrations of human chemokines in the supernatant including C-C-motif ligand 1 (CCL1), CCL2, CCL4, CCL20, C-XC-motif ligand 1 (CXCL1), CXCL2, CXCL3, CXCL8, CXCL16, and C-motif ligand 1 (XCL1) were measured using human ELISA kits from CUSABIO for CCL1 (#CSB-EL004774HU), CCL2 (#CSB-Eq.

Techniques: Cell Counting, Blocking Assay, Flow Cytometry, Fluorescence

Journal: Virology Journal

Article Title: The CC chemokine ligand (CCL) 1, upregulated by the viral transactivator Tax, can be downregulated by minocycline: possible implications for long-term treatment of HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/s12985-017-0902-6

Figure Lengend Snippet: Preferential expression of CCL1 in Human T-cell leukemia virus type-1 (HTLV-1)-infected T-cell lines derived from patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). a . Expression of CCL1 was examined by RT-PCR in HTLV-1-infected and -uninfected T-cell lines. CCL1 mRNA was preferentially expressed in HTLV-1-infected human T-cell lines derived from patients with HAM/TSP (4 out of 4 tested), compared with HTLV-1-transformed T-cell lines (1 out of 3) and adult T-cell leukemia (ATL) cell lines (1 out of 4). b . The expressions of CCL1 , tax , and HBZ were examined by real time PCR in HTLV-1-infected and -uninfected T-cell lines. CCL1 mRNA was preferentially expressed in HTLV-1-infected human T-cell lines derived from patients with HAM/TSP. The expression levels of the viral RNAs tax and HBZ were relatively high in T-cell lines derived from patients with HAM/TSP and HTLV-1-transformed T-cell lines when compared with those in ATL cell lines. c . CCL1 levels in culture supernatants from HTLV-1-infected and -uninfected human T-cell lines were assessed by ELISA. Supernatants were harvested when cells reached subconfluency. Significant levels of CCL1 was observed in the culture supernatants from HTLV-1-infected human T-cell lines derived from patients with HAM/TSP, whereas it was not detectable in any of the other cell lines tested except for the HTLV-1-transformed C5MJ cell line

Article Snippet: Human CCL1 gene-specific assays (Applied Biosystems Hs00171072 m1) were used for CCL1 quantification.

Techniques: Expressing, Virus, Infection, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Transformation Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Virology Journal

Article Title: The CC chemokine ligand (CCL) 1, upregulated by the viral transactivator Tax, can be downregulated by minocycline: possible implications for long-term treatment of HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/s12985-017-0902-6

Figure Lengend Snippet: Tax-dependent constitutive expression of CCL1 in Tax-inducible JPX9 cells. RNA and protein expression of CCL1 in JPX9 cells. JPX9 is a Jurkat (HTLV-1-negative human T-cell leukemia cell line) subclone generated by stable transfection of a functional Tax expression-plasmid vector. Tax expression was induced by adding CdCl 2 to the culture medium (final concentration: 10 μM). RT-PCR ( a ) and western blotting ( b ) showed that treatment of JPX9 cells with CdCl 2 induced the expression of Tax at the RNA and protein levels, respectively. c . Flow cytometry data showed that treatment of JPX9 cells with CdCl 2 induced CCL1 expression and that CCL1 was expressed exclusively in cells that also expressed Tax. d . ELISA analysis of culture supernatants showed that the addition of CdCl 2 to the JPX9 cell culture resulted in an increase in CCL1 expression within 24 h. CCL1 levels were found to reach a peak after 48 h of culture

Article Snippet: Human CCL1 gene-specific assays (Applied Biosystems Hs00171072 m1) were used for CCL1 quantification.

Techniques: Expressing, Generated, Stable Transfection, Functional Assay, Plasmid Preparation, Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Virology Journal

Article Title: The CC chemokine ligand (CCL) 1, upregulated by the viral transactivator Tax, can be downregulated by minocycline: possible implications for long-term treatment of HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/s12985-017-0902-6

Figure Lengend Snippet: HTLV-1 Tax transactivates the CCL1 gene by acting concurrently on AP-1- and CREB-binding sites. The luciferase gene was used to monitor the activity of the CCL1 gene promoter. Four fragments of the 5′-flanking regulatory region of the CCL1 gene were cloned upstream of the luciferase gene, yielding four constructs. These reporter constructs were independently transfected into Jurkat human T-cells with or without the Tax expression plasmid. The results showed that both AP-1- and CREB-binding sites were essential and sufficient for transactivation of the CCL1 gene by HTLV-1 Tax

Article Snippet: Human CCL1 gene-specific assays (Applied Biosystems Hs00171072 m1) were used for CCL1 quantification.

Techniques: Binding Assay, Luciferase, Activity Assay, Clone Assay, Construct, Transfection, Expressing, Plasmid Preparation

Journal: Virology Journal

Article Title: The CC chemokine ligand (CCL) 1, upregulated by the viral transactivator Tax, can be downregulated by minocycline: possible implications for long-term treatment of HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/s12985-017-0902-6

Figure Lengend Snippet: Plasma levels of CCL1 were significantly higher in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) than in HTLV-1-seronegative patients with multiple sclerosis (MS) and asymptomatic HTLV-1 healthy carriers (HC). The plasma levels of CCL1 in 31 patients with HAM/TSP, 13 HTLV-1-seronegative patients with MS, 19 HCs, and 10 normal controls (NCs) were examined by ELISA. As shown, the plasma levels of CCL1 were significantly higher in patients with HAM/TSP (mean ± SD: 79.3 ± 30.5 pg/ml) than in HTLV-1-seronegative patients with MS (36.8 ± 11.3 pg/ml), HCs (28.7 ± 2.5 pg/ml), and NCs (28.4 ± 3.2 pg/ml)

Article Snippet: Human CCL1 gene-specific assays (Applied Biosystems Hs00171072 m1) were used for CCL1 quantification.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Virology Journal

Article Title: The CC chemokine ligand (CCL) 1, upregulated by the viral transactivator Tax, can be downregulated by minocycline: possible implications for long-term treatment of HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/s12985-017-0902-6

Figure Lengend Snippet: Minocycline inhibits the production of CCL1 in HTLV-1-infected T-cell lines. Effect of minocycline on CCL1 protein expression in the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP)-derived HTLV-1-infected T-cell line ILT-M1 and in the HTLV-1-transformed cell line C5MJ. Minocycline suppressed the expression of CCL1 in both ILT-M1 and C5MJ cell lines in a dose-dependent manner

Article Snippet: Human CCL1 gene-specific assays (Applied Biosystems Hs00171072 m1) were used for CCL1 quantification.

Techniques: Infection, Expressing, Derivative Assay, Transformation Assay

Journal: The Journal of Experimental Medicine

Article Title: STARTRAC analyses of scRNAseq data from tumor models reveal T cell dynamics and therapeutic targets

doi: 10.1084/jem.20201329

Figure Lengend Snippet: CCR8 expression in tumor T reg cells and characterization of anti-CCR8 antibodies . (A) Representative histogram plot depicting expression of CCR8 on 4-1BB + and 4-1BB − CD4 + Foxp3 − T EFF population in MC38 tumors. (B) Representative histogram plot depicting expression of CCR8 on CCR7 + and CCR7 − CD8 + T cells in MC38 tumors. (C) Flow cytometry analysis of tumor-infiltrating T reg cells across multiple mouse tumor models shown as representative FACS plots (left) and frequencies in tumors and peripheral lymphoid organs (right). *, P = 0.0250; ****, P < 0.0001; two-way ANOVA with Tukey’s post-hoc analysis. (D) Flow cytometry analysis of CCR8 expression on immune cell types (Foxp3 + T reg cells, Foxp3 − T effectors, CD8 + , NKp46 + NK, CD19 + B, and CD11b + ) in tumors and peripheral lymphoid tissues across tumor models. (E) In vitro chemotaxis assay evaluating the ability of CCR8-depleting and -nondepleting antibodies to block 100 pM CCL1-induced chemotaxis. (F) Bar graph comparing IC 50 values from the chemotaxis assay testing CCR8-depleting and -nondepleting antibodies ( n = 6). (G) Pharmacokinetic assessment of serum antibody concentration after single-dose administration of CCR8-depleting and -nondepleting antibodies (10 mg/kg i.p.). (H) Serum concentration at 72 h of animals treated as in G. DLN, draining LN; SPL, spleen; MFI, mean fluorescence intensity.

Article Snippet: Recombinant human CCL1 (R&D Systems) was prepared at suboptimal concentration of 100 pM and added to the bottom Transwell chambers at 100 μl per well.

Techniques: Expressing, Flow Cytometry, In Vitro, Chemotaxis Assay, Blocking Assay, Concentration Assay, Fluorescence