Journal: British Journal of Cancer

Article Title: Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis

doi: 10.1038/bjc.2013.565

Figure Lengend Snippet: CX3CR1 and CX3CL1 expression in normal pancreas and in pancreatic cancer. ( A ) CX3CR1 expression in normal pancreas, ( B ) in well- and ( C ) in poorly differentiated PDAC, and ( B , inset) in inflammatory cells in tumour microenvironment. ( D ) CX3CL1 expression in normal pancreas, ( E ) in neural cell bodies, and ( F ) in PDAC (box, × 40 magnification of a CX3CL1 + PDAC). Objective magnification × 10 ( A – D and F ); ( E ) and inset in ( B ), × 40. ( G ) Frequency of CX3CR1 + and CX3CR1 − PDAC with PNI according to tumour differentiation (Grade, G1–G2 well-to-moderately differentiated PDAC; G3, poorly differentiated PDAC). ( H ) Kaplan–Meier curves for overall survival of patients after radical resection of PDAC ( n =67), according to the status of CX3-CR1 (left panel) and -CL1 (right panel) in cancer. P -values by log-rank test. * indicates P <0.05.

Article Snippet: After deparaffining and rehydration, the sections were immersed in antigen retrieval bath, incubated with 3% H 2 O 2 for 15 min, and treated for 2 h at room temperature with primary antibodies raised against CX3CR1 (Ab8021, polyclonal rabbit anti-human; Abcam, Cambridge, UK) and CX3CL1 (AF365, polyclonal goat anti-human; R&D Systems, Minneapolis, MN, USA), or with rabbit or goat IgG (Dako, Milan, Italy) to serve as negative controls, followed by 30-min incubation with the DAKO Envision system (Dako) or the Anti-Goat Polymer kit (Biocare, San Francisco, CA, USA).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis

doi: 10.1038/bjc.2013.565

Figure Lengend Snippet: CX3CR1 and CX3CL1 expression in 104 PDAC according to patients' demographics and to tumour features

Article Snippet: After deparaffining and rehydration, the sections were immersed in antigen retrieval bath, incubated with 3% H 2 O 2 for 15 min, and treated for 2 h at room temperature with primary antibodies raised against CX3CR1 (Ab8021, polyclonal rabbit anti-human; Abcam, Cambridge, UK) and CX3CL1 (AF365, polyclonal goat anti-human; R&D Systems, Minneapolis, MN, USA), or with rabbit or goat IgG (Dako, Milan, Italy) to serve as negative controls, followed by 30-min incubation with the DAKO Envision system (Dako) or the Anti-Goat Polymer kit (Biocare, San Francisco, CA, USA).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis

doi: 10.1038/bjc.2013.565

Figure Lengend Snippet: Tumour differentiation, peri-neural invasion, and ligand expression are independently associated with CX3CR1 expression

Article Snippet: After deparaffining and rehydration, the sections were immersed in antigen retrieval bath, incubated with 3% H 2 O 2 for 15 min, and treated for 2 h at room temperature with primary antibodies raised against CX3CR1 (Ab8021, polyclonal rabbit anti-human; Abcam, Cambridge, UK) and CX3CL1 (AF365, polyclonal goat anti-human; R&D Systems, Minneapolis, MN, USA), or with rabbit or goat IgG (Dako, Milan, Italy) to serve as negative controls, followed by 30-min incubation with the DAKO Envision system (Dako) or the Anti-Goat Polymer kit (Biocare, San Francisco, CA, USA).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis

doi: 10.1038/bjc.2013.565

Figure Lengend Snippet: Expression of CX3CR1 and CX3CL1 in precursor lesions according to tumour differentiation and PanIN degree. ( A ) Comparison of the expression rate of CX3CR1 (upper panels) and CX3CL1 (lower panels) in paired precursor lesions and invasive cancers within the same tissue specimens ( n =76), according to the degree of pancreatic cancer differentiation. Number within bars, number of specimens. ( B ) Rates of CX3CR1 + (upper panels) and CX3CL1 + (lower panels) PanINs according to their degree of dysplasia. Numbers within bars, number of PanINs. G1–G2 tumours, left panels; G3 tumours, right panels; P -values by Fisher's exact test.

Article Snippet: After deparaffining and rehydration, the sections were immersed in antigen retrieval bath, incubated with 3% H 2 O 2 for 15 min, and treated for 2 h at room temperature with primary antibodies raised against CX3CR1 (Ab8021, polyclonal rabbit anti-human; Abcam, Cambridge, UK) and CX3CL1 (AF365, polyclonal goat anti-human; R&D Systems, Minneapolis, MN, USA), or with rabbit or goat IgG (Dako, Milan, Italy) to serve as negative controls, followed by 30-min incubation with the DAKO Envision system (Dako) or the Anti-Goat Polymer kit (Biocare, San Francisco, CA, USA).

Techniques: Expressing, Comparison

Journal: British Journal of Cancer

Article Title: Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis

doi: 10.1038/bjc.2013.565

Figure Lengend Snippet: CX3CR1 and CX3CL1 expression in precursor lesions of pancreatic cancer. ( A ) Left panels, human PanINs; right panels, PanINs of PdxCre/LSL- Kras G12D mice. Objective magnification, × 20 (human PanINs), and × 40 (mice PanINs). ( B ) Precursor lesions adjacent to neural structures. Detection of precursor lesions close to (left panel) or in tight contact with (right panel) S-100 + neural structures (red arrows) in pancreata of 6-month-old PdxCre/LSL-Kras G12D mice. Magnification, × 20.

Article Snippet: After deparaffining and rehydration, the sections were immersed in antigen retrieval bath, incubated with 3% H 2 O 2 for 15 min, and treated for 2 h at room temperature with primary antibodies raised against CX3CR1 (Ab8021, polyclonal rabbit anti-human; Abcam, Cambridge, UK) and CX3CL1 (AF365, polyclonal goat anti-human; R&D Systems, Minneapolis, MN, USA), or with rabbit or goat IgG (Dako, Milan, Italy) to serve as negative controls, followed by 30-min incubation with the DAKO Envision system (Dako) or the Anti-Goat Polymer kit (Biocare, San Francisco, CA, USA).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis

doi: 10.1038/bjc.2013.565

Figure Lengend Snippet: Effects of microenvironmental stimuli on CX3CR1 and CX3CL1 expression in pancreatic tumoral cells. CX3CR1 (left panels) and CX3CL1 (right panels) mRNA fold changes induced by stimulation with TNF- α +IFN- γ , IL-1 β , IL-6, and by TGF- β in mouse DT6606 ( PdxCre/LSL-Kras G12 model) and K8484 ( PdxCre/LSL - Kras G12D - Trp53 R172H model) cells, and in human A8184, AspCI, and MiaPacaII pancreatic cancer cells. * indicates P <0.05.

Article Snippet: After deparaffining and rehydration, the sections were immersed in antigen retrieval bath, incubated with 3% H 2 O 2 for 15 min, and treated for 2 h at room temperature with primary antibodies raised against CX3CR1 (Ab8021, polyclonal rabbit anti-human; Abcam, Cambridge, UK) and CX3CL1 (AF365, polyclonal goat anti-human; R&D Systems, Minneapolis, MN, USA), or with rabbit or goat IgG (Dako, Milan, Italy) to serve as negative controls, followed by 30-min incubation with the DAKO Envision system (Dako) or the Anti-Goat Polymer kit (Biocare, San Francisco, CA, USA).

Techniques: Expressing

Journal: JCI Insight

Article Title: The homozygous CX3CR1-M280 mutation impairs human monocyte survival

doi: 10.1172/jci.insight.95417

Figure Lengend Snippet: Percent of dead CD14+ monocytes elicited by serum starvation in the presence or absence of 100 nM of soluble or full-length CX3CL1 in CX3CR1-WT/WT monocytes. Monocytes cultured in serum-containing media were assayed as control. n = 6. ***P < 0.001; ****P < 0.0001. Statistical analysis was performed using 1-way ANOVA with Tukey’s multiple comparisons test. Data represent the mean ± SEM.

Article Snippet: CD14 + monocytes (1 × 10 5 ) from CX3CR1-WT/WT , heterozygous CX3CR1-WT/M280 , and homozygous CX3CR1-M280/M280 donors were serum starved in 5 ml polystyrene round-bottom tubes (BD Falcon) by resuspension in either RPMI 1640 containing 100 U/ml of penicillin and 100 μg/ml of streptomycin or RPMI 1640 containing antibiotics and 100 nM final concentration of recombinant human full-length CX3CL1 (R&D Systems, catalog 365-FR) or 100 nM final concentration of recombinant soluble CX3CL1 (R&D Systems, catalog 362-CX-025) at 37°C for 4 hours.

Techniques: Cell Culture, Control

Journal: JCI Insight

Article Title: The homozygous CX3CR1-M280 mutation impairs human monocyte survival

doi: 10.1172/jci.insight.95417

Figure Lengend Snippet: (A) Representative FACS histograms of propidium iodine (PI) staining in CX3CR1-WT/WT (upper panels) and CX3CR1-M280/M280 (lower panels) CD14+ monocytes following serum starvation in the presence or absence of 100 nM of CX3CL1. (B) Percent of dead CD14+ monocytes elicited by serum starvation in the presence or absence of 100 nM of CX3CL1 in CX3CR1-WT/WT (left panel), CX3CR1-WT/M280 (middle panel), and CX3CR1-M280/M280 (right panel) cells. Shown are paired experimental results with or without CX3CL1. (C) The percent decrease in cell death conferred by CX3CL1 exposure in serum-starved CD14+ monocytes is greater in CX3CR1-WT/WT compared with CX3CR1-WT/M280 cells, while no CX3CL1-induced decrease in cell death is seen in CX3CR1-M280/M280 cells. n = 22 CX3CR1-WT/WT, 11 CX3CR1-WT/M280 and 6 CX3CR1-M280/M280. *P < 0.05; ***P < 0.001; ****P < 0.0001. Statistical analysis was performed using paired 2-tailed t tests (B) or 1-way ANOVA with Tukey’s multiple comparisons test (C). Quantitative data represent the mean ± SEM.

Article Snippet: CD14 + monocytes (1 × 10 5 ) from CX3CR1-WT/WT , heterozygous CX3CR1-WT/M280 , and homozygous CX3CR1-M280/M280 donors were serum starved in 5 ml polystyrene round-bottom tubes (BD Falcon) by resuspension in either RPMI 1640 containing 100 U/ml of penicillin and 100 μg/ml of streptomycin or RPMI 1640 containing antibiotics and 100 nM final concentration of recombinant human full-length CX3CL1 (R&D Systems, catalog 365-FR) or 100 nM final concentration of recombinant soluble CX3CL1 (R&D Systems, catalog 362-CX-025) at 37°C for 4 hours.

Techniques: Staining

Journal: JCI Insight

Article Title: The homozygous CX3CR1-M280 mutation impairs human monocyte survival

doi: 10.1172/jci.insight.95417

Figure Lengend Snippet: Shown are percent of dead monocytes elicited by serum starvation in the presence or absence of 100 nM of CX3CL1 after exposure to 50 μM of the PI3K inhibitor LY294002 (A, n = 5) or 20 μM of the MEK inhibitor PD98059 (B, n = 6), or 100 nM of the PI3K inhibitor wortmannin (C, n = 6) or 20 μM of the MEK inhibitor U0126 (D, n = 5). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Statistical analysis was performed using 1-way ANOVA with Tukey’s multiple comparisons test. Data represent the mean ± SEM.

Article Snippet: CD14 + monocytes (1 × 10 5 ) from CX3CR1-WT/WT , heterozygous CX3CR1-WT/M280 , and homozygous CX3CR1-M280/M280 donors were serum starved in 5 ml polystyrene round-bottom tubes (BD Falcon) by resuspension in either RPMI 1640 containing 100 U/ml of penicillin and 100 μg/ml of streptomycin or RPMI 1640 containing antibiotics and 100 nM final concentration of recombinant human full-length CX3CL1 (R&D Systems, catalog 365-FR) or 100 nM final concentration of recombinant soluble CX3CL1 (R&D Systems, catalog 362-CX-025) at 37°C for 4 hours.

Techniques:

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Chemokines and chemokine receptors expression in the lesions of patients with American cutaneous leishmaniasis

doi: 10.1590/s0074-0276108042013008

Figure Lengend Snippet: Fig. 1: chemokines and chemokine receptors in lesions of different clinical forms of American cutaneous leishmaniasis. A, B: density of CXCL10, CCL4, CXCR3 and CCR5 positive cells; C, D: density of CCL11, CCL8 and CCR3 positive cells; E: density of CXCL8; F: CCR7 posi tive cells. Data are represented as medians and interquartile range patients with localised cutaneous leishmaniasis (LCL) (n = 20), intermediate cutaneous leishmaniasis (ICL) (n= 5) and diffuse cutaneous leishmaniasis (DCL) (n = 10). Asterisk means p < 0.05 comparing LCL with DCL, LCL with ICL and LCL with ICL by the Mann-Witney U test.

Article Snippet: Mouse monoclonal antibodies against the following human chemokine receptors were purchased from R&D Systems Inc (Minneapolis, USA) (5 μg/mL): CXCR3 (MAB-160), CCR3 (MAB-155), CCR5 (MAB-181) and CCR7 (MAB-197).

Techniques:

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Chemokines and chemokine receptors expression in the lesions of patients with American cutaneous leishmaniasis

doi: 10.1590/s0074-0276108042013008

Figure Lengend Snippet: Fig. 2: immunolabelling of chemokine in lesions American cutane ous leishmaniasis. Detection of CXCL10 on the epidermal cells (A), nerve fibres (B) and glands (C) of a patient with localised cutaneous leishmaniasis (LCL) with their respective negative controls (D, E, F). Epidermal CCL11+ dendritic cells (D) in a patient with diffuse cuta neous leishmaniasis (DCL). Infiltrating CCR3+ cells (E) in a patient with DCL. CCR7+ cells in a patient with LCL (F). Respective nega tive controls (J-L). Original magnification: 400X and 200X (A, D).

Article Snippet: Mouse monoclonal antibodies against the following human chemokine receptors were purchased from R&D Systems Inc (Minneapolis, USA) (5 μg/mL): CXCR3 (MAB-160), CCR3 (MAB-155), CCR5 (MAB-181) and CCR7 (MAB-197).

Techniques: