Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: Characterization of CCL2 -expressing clones. CCL2 ELISA results for a positive control (RH-6), a negative control (OV-90), stable EV clones: OV-90:EV 2 , OV-90:EV 9 , and stable CCL2 -expressing clones: OV-90: CCL2 23 and OV-90: CCL2 49 ( a ). Semi-quantitative RT–PCR analysis of CCL2 performed on complementary (c) DNA samples prepared from the parental OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 ( b ). The expression of 18S cDNA is shown for RNA quality. Light microscope photographs of the cell morphology of OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 , arrows indicate representative cell morphology ( c ). Light microscope photographs are × 100 magnification. Cell viability by XTT assay of OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 , representative results from one independent experiment that was performed in triplicate ( d ). Cell viability was measured over 8 days and cell culture media-only wells served as controls (OSE and OSE-G418).

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Expressing, Clone Assay, Enzyme-linked Immunosorbent Assay, Positive Control, Negative Control, Quantitative RT-PCR, Light Microscopy, XTT Assay, Cell Culture

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: In vitro characterization of the CCL2 -expressing clones. Light microscope photographs of OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 spheroids on day 8 of the assay, edges of the hanging droplets are shown ( a ). Colony forming ability of OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 ( b ). Magnified ( × 4) colonies formed by OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 ( c ). Average number of colonies formed in colony formation assay, representative results from one independent experiment that was performed in triplicate ( d ). Wound healing assay of OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 and OV-90: CCL2 49 over a period of 5 days ( e ). Light microscope photographs are × 100 magnification.

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: In Vitro, Expressing, Clone Assay, Light Microscopy, Colony Assay, Wound Healing Assay

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: S.c. injection site in vivo tumourigenicity assay. Mean survival, in days, of mice injected with OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 or OV-90: CCL2 49 at s.c. injection sites with six mice per group, and four phosphate-buffered saline (PBS)-injected mice as injection controls ( a ). Final mean tumour volume per group ( b ). Mean tumour volume per group over time ( c ).

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Injection, In Vivo, Saline

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: I.p. injection site in vivo tumourigenicity assay. Mean survival, in days, of mice injected with OV-90, OV-90:EV 2 , OV-90:EV 9 , OV-90: CCL2 23 or OV-90: CCL2 49 at i.p. injection sites with six mice per group, and four PBS-injected mice as injection controls ( a ). Kaplan–Meier survival curves comparing all groups ( b ).

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Injection, In Vivo

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: CCL2 expression and hematoxylin and eosin staining of xenografts. Semi-quantitative RT–PCR analysis of endogenous CCL2 ( CCL2 ) and introduced, expression vector-derived, CCL2 ( Flag/CCL2 ) performed on cDNA samples prepared from a subset of xenografts obtained from both i.p. and s.c. injection sites ( a ). The expression of 18S cDNA is shown for RNA quality. Hematoxylin and eosin staining of OV-90, OV-90: CCL2 23 and OV-90: CCL2 49 xenografts obtained from both s.c. and i.p. injection sites at time of killing ( b ). Images were obtained from the OlyVIA image viewer (Olympus America Inc., Center Valley, PA, USA).

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Expressing, Staining, Quantitative RT-PCR, Plasmid Preparation, Derivative Assay, Injection

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: Enriched biological term and biological function analysis of the 89 probe sets differentially expressed in the CCL2 -expressing clones. Significantly enriched GO biological process terms derived from the DAVID functional annotation classification tool, with P -values <0.05 classified as significant by a modified Fisher's exact test ( a ). Ingenuity pathway analysis of enriched biological functions ( b ).

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Expressing, Clone Assay, Derivative Assay, Functional Assay, Modification

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: CCL2 expression in HGOSC. CCL2 expression in 11 NOSE samples, normal whole ovary and 79 HGOSC samples as determined by Ziplex Research System expression array (Axela Inc., Toronto, Ontario, Canada) ( a ). Examples of CCL2 expression in cores from four HGOSC samples (left panel) showing, negative, detectable but low, moderately detectable and clearly detectable CCL2 staining, and examples of CCL2 staining in cores from two normal fallopian tube tissue samples (right panel) ( b ). Percentages refer to the proportion of the 187 HGOSC samples showing negative, detectable but low, moderately detectable and clearly detectable staining patterns of protein expression . ‘E' indicates epithelial cells and ‘S' indicates stromal cells. Immunohistochemistry images were obtained from the OlyVIA image viewer (Olympus America Inc.). The Kaplan–Meier survival curve analysis of HGOSC cases for overall ( c ) and disease-free ( d ) survival (in months) of patients whose tumours showed negative or low CCL2 staining ( n =136) compared with patients whose tumours showed moderately or clearly detectable CCL2 staining ( n =51). Indicated P -values were derived from log-rank tests.

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Expressing, Staining, Immunohistochemistry, Derivative Assay

Journal: Oncogenesis

Article Title: Overexpressing the CCL2 chemokine in an epithelial ovarian cancer cell line results in latency of in vivo tumourigenicity

doi: 10.1038/oncsis.2012.25

Figure Lengend Snippet: Summary of immunohistochemistry analysis of CCL2 in HGOSC samples

Article Snippet: Staining was performed with an antibody against human CCL2 (1 : 100; R&D Systems, Burlington, Ontario, Canada), using the Ventana Benchmark XT system (Ventana Medical Systems, Inc., Tucson, Arizona).

Techniques: Immunohistochemistry, Staining

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 1. Exposure to high altitude results in PH and increased secretion of inflammatory classical monocyte ligands from the lungs. (A) Schematic showing hypoxia exposure time course in wildtype mice. Duration of hypoxia exposure is directly proportional to (B) RVSP and RV hypertrophy as measured by Fulton Index (N=6-13/group). At 3 days of hypoxia, increased protein expression of classical monocyte ligands (C) CCL2 (N=6-11/group) and (D) CCL12 (N=6- 11/group), whereas significantly lower levels of nonclassical monocyte ligand (E) CX3CL1 (N=6/group) in the lungs. (F) Higher CCL2 gradient in lungs and in the (G) peripheral blood of wildtype mice following 3 days of hypoxia exposure (N=5/group). Data in all panels were obtained from female mice. Statistical analysis was conducted using ANOVA, followed by Tukey's post hoc test. *P<0.05, **P<0.01, ****P<0.0001. N=number of animals, mean±SD, CI=confidence interval.

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Expressing

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 4: Genetic and pharmacologic blockade of CCR2-CCL2 axis protects from hypoxic PH. (A) Schematic showing the BM reconstitution of Ccr2-/- and WT BM into lethally irradiated wildtype mice. Wildtype mice reconstituted with Ccr2-/- BM were protected from hypoxic PH by attenuated (B) RVSP (N=7-11/group) and (C) RV hypertrophy (N=7-11/group) as measured by Fulton Index, compared to wildtype mice that were reconstituted with wildtype BM. (D) Schematic showing pharmacological blockade of CCR2 ligands CCL2 or CCL7 using anti-CCL2 or anti-CCL7 neutralizing antibody treatment. Hypoxia exposed wildtype mice treated with CCL2 NAb but not CCL7 NAb showed lower (E) RVSP (N=6/group) and (F) RV hypertrophy (N=6/group). TSP-1 levels in (G) lungs (N=6/group) and (H) blood (N=6/group); and TGF-β1 levels in (I) lungs (N=6/group) and (J) blood (N=6/group) compared to wildtype mice treated with isotype control antibody. Data in all panels followed a normal distribution. ANOVA with the Tukey test was performed for multiple comparisons. Data were obtained from the female mice. mean ± SD

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Irradiation, Control

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 5: Resident IMs are a major source of CCL2 and recruited IMs are a major source of pathologic TSP-1 in hypoxic PH. (A) Flow cytometry analysis using Ccl2RFP-flox reporter mice showed a higher number of CCL2+ IMs (N=14/group; N=14/group, 9F and 5M in Nx; 8F and 6M in Hx), and (B) FOLR2+ IMs are a major source of CCL2 (N=14/group). (C) Hypoxia exposed wildtype mice following intracellular CCL2 staining by flow cytometry also showed a higher number of CCL2+ IMs (N=7/group, female mice). (D). IM subpopulation analysis using flow

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Flow Cytometry, Staining

Journal: Journal of Clinical Investigation

Article Title: Monocytes and interstitial macrophages contribute to hypoxic pulmonary hypertension

doi: 10.1172/jci176865

Figure Lengend Snippet: Figure 8: DEX prophylaxis blunts CCL2 production by resident IMs and blocks the recruitment of TSP-1 producing CCR2+ IMs in hypoxia. (A) DEX prophylactically-treated, hypoxia-exposed Ccl2RFP-flox reporter mice exhibited a significant reduction in CCL2+ IMs, particularly in (B) CCL2RFP+ resident IMs (N=7/group). Additionally, (C) intracellular CCL2 flow cytometry analysis in DEX prophylactically-treated hypoxia-exposed wildtype mice revealed a

Article Snippet: Neutralizing Antibody and Pharmacological Treatment: Neutralizing mouse antibodies against CCL2 (Clone: 2H5; Cat.# BE0185), CCL7 (R&D System; Cat.# AF-456-NA), and isotype control (Cat.# BE0091; BioXCell, West Lebanon, NH, USA) were reconstituted in phosphatebuffered saline (PBS).

Techniques: Flow Cytometry

Journal: Biomedicines

Article Title: Regulation of Cancer Stem Cells and Epithelial-Mesenchymal Transition by CTNNAL1 in Lung Cancer and Glioblastoma.

doi: 10.3390/biomedicines11051462

Figure Lengend Snippet: Figure 4. Effects of CCL2 regulated by CTNNAL1 on lung cancer cells. (A) Cytokine analysis using cytokine array. A549 cells were transfected with siRNA to knock down the CTNNAL1 gene. (B) Determination of CCL2 expression at the gene level. The expression of CTNNAL1 was knocked down using siRNA in A549 cells. (C) After treatment with CCL2 neutralizing antibody, the expression of CTNNAL1 was confirmed. (D) After treatment with CCL2 neutralizing antibodies, the expression of CSC marker proteins CD44, ALDH1A1, and ALDH1A3 was confirmed by WB. (E) Comparative analysis of EMT marker proteins E-cadherin, N-cadherin, and Vimentin after treatment with CCL2 neutralizing antibody.

Article Snippet: Antibody CCL2 (2 μg/mL; cat. no. MAB679; R&D SYSTEMS, Minneapolis, MN, USA) and normal mouse IgG1 antibody (1 μg/mL; cat. no. sc-3877; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) were used in the neutralization assay.

Techniques: Transfection, Knockdown, Expressing, Marker

Journal: Journal of Biological Chemistry

Article Title: Hepatocyte nuclear receptor SHP suppresses inflammation and fibrosis in a mouse model of nonalcoholic steatohepatitis

doi: 10.1074/jbc.ra117.001653

Figure Lengend Snippet: FIGURE 3. SHP level is decreased in the livers of mice NASH. A, qPCR analysis of relative mRNA levels of genes related to lipid metabolism, inflammation, and fibrosis in the livers of mice fed chow or HFCF for 1 and 5 moths. N=5 mice/Group. *p < 0.05 HFCF vs. Chow. B, Serum level of CCL2 was measured by enzyme-linked immunosorbent assay (ELISA). N=5 mice/Group. *p < 0.05 HFCF vs. Chow. C, qPCR analysis of Shp mRNA level in the livers of mice fed chow or HFCF. N=5 mice/Group. *p < 0.05 HFCF vs. Chow. D, Left: Western blotting analysis of SHP protein in the livers of mice fed chow or HFCF for 5 months. SHP (H-160) is a rabbit polyclonal antibody and SHP (H-5) is a mouse monoclonal antibody. Both antibodies recognize the epitope corresponding to amino acids 1-160 mapping at the N-terminus of SHP protein. Right: Band intensities were calculated using Image J software. The level of SHP was normalized to the expression of loading control β-actin and fold changes relative to that of the controls are plotted. N=5 mice/Group. *p < 0.05 HFCF vs. Chow. E, Representative images of liver sections stained with hematoxylin-eosin (H&E) in mice fed chow or methionine-choline deficient (MCD) diet for 1 month. Original magnification, x40. F, qPCR analysis of gene expression in the livers of mice fed chow or MCD diet for 1 month. N=5 mice/Group. Data are presented as mean ± SD. *p < 0.05 versus respective controls.

Article Snippet: Recombinant mouse CCL2 protein (479-JE-010) and anti-mouse CCL2 antibody (AF-479-SP) for neutralization were obtained from RD system.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Software, Expressing, Control, Staining, Gene Expression

Journal: Frontiers in Immunology

Article Title: Directing the migration of serum-free, ex vivo -expanded Vγ9Vδ2 T cells

doi: 10.3389/fimmu.2024.1331322

Figure Lengend Snippet: γδ T-cell homing can be influenced by leveraging the CCR2/CCL2 axis. (A) Flow cytometry histograms depicting CCR2 expression on γδ T cells with two different CCR2 antibody clones; 7-AAD was used as a live/dead control. (B) Representative human chemokine array membrane blot surveying secretion by MSCs from MSC- or γMSC-conditioned media. (C) Graphical depiction of the transwell migration assay with γδ T cells migrating to either MSCs or γMSCs with the respective conditioned media. (D) Specific migration of γδ T cells to MSCs or γMSCs in the transwell assay; n = 3 biological replicates, statistics analyzed by Student’s t-test (p< 0.05 = *), and error bars represent standard deviation. MSCs, mesenchymal stromal cells.

Article Snippet: To confirm that CCL2 induced γδ T-cell migration to γMSCs, a transwell migration assay was performed using a CCL2 antibody (R&D Systems, Minneapolis, MN, USA; MAB679-500) blockade.

Techniques: Flow Cytometry, Expressing, Clone Assay, Control, Membrane, Transwell Migration Assay, Migration, Transwell Assay, Standard Deviation

Journal: Molecular human reproduction

Article Title: IL-33 enhances proliferation and invasiveness of decidual stromal cells by up-regulation of CCL2/CCR2 via NF-κB and ERK1/2 signaling.

doi: 10.1093/molehr/gat094

Figure Lengend Snippet: Figure 4 Effects of IL-33/ST2L/sST2 axis on CCL2/CCR2 expres- sion in DSCs. The secretion of chemokine CCL2 (A) by DSCs was determined by ELISA, and the expression of CCR2 on DSCs (B) by flow cytometry. DSCs (2 × 105 cells/well) were seeded in 24-well plates with or without rhIL-33(0.01–10 ng/ml) and sST2 (100– 400 ng/ml). The supernatant was then collected after 48 h while DSCs were used to detect the protein concentration, and the CCL2 levelofeachgroupwascalculatedastheratiooftheCCL2concentration of supernatant to the protein concentration. The pictures are from a representative experiment and the numbers are the percentages of positive cells. Results are representative of three independent experi- ments. ***P , 0.001. CCL2-APC, allophycocyanin labeled CCL2 antibody.

Article Snippet: BrdU cell proliferation assay Bromodeoxyuridine (5-bromo-2′-deoxyuridine, BrdU) cell proliferation assay was applied to evaluate the effect of IL-33 on cell proliferation with or without sST2, anti-CCL2 neutralizing antibody (MAB279, R&D Systems) and CCR2 blocker RS102895 (R1903, Sigma-Aldrich, Santa Clara, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Cytometry, Protein Concentration, Labeling

Journal: Molecular human reproduction

Article Title: IL-33 enhances proliferation and invasiveness of decidual stromal cells by up-regulation of CCL2/CCR2 via NF-κB and ERK1/2 signaling.

doi: 10.1093/molehr/gat094

Figure Lengend Snippet: Figure 5 The increased proliferation and invasiveness of DSCs induced by IL-33 are dependent on CCL2/CCR2 interaction. DSCs (1 × 104 cell/well, BrdUproliferationassay;3 × 105 cells/well,cellcycleassay;2 × 105 cells/well,invasionassay)weretreatedwithrhIL-33(1 ng/ml),a-CCL2(1 mg/ml),or CCR2 blocker RS102895(100 ng/ml) with vehicle as control. After stimulation with rhIL-33 (1 ng/ml), the cells were treated with CCL2 neutralizing anti- body (1 mg/ml) or RS102895 (100 ng/ml) foranother 48 h. Thereafter, BrdU proliferation assay(A), cell cycle (B) and invasion assay (C) wereconducted to analyze proliferation and invasiveness of DSCs, respectively. Error bars depict the standard deviation of the sample mean. The images are from an ex- periment that is representative of three independently conducted experiments. The x-axis, from the left, represents the cell cycle phases of G1(green region), S(the region between G1 and G2) and G2(blue region). *P , 0.05, **P , 0.01, ***P , 0.001.

Article Snippet: BrdU cell proliferation assay Bromodeoxyuridine (5-bromo-2′-deoxyuridine, BrdU) cell proliferation assay was applied to evaluate the effect of IL-33 on cell proliferation with or without sST2, anti-CCL2 neutralizing antibody (MAB279, R&D Systems) and CCR2 blocker RS102895 (R1903, Sigma-Aldrich, Santa Clara, USA).

Techniques: Control, Proliferation Assay, Invasion Assay, Standard Deviation

Journal: Molecular human reproduction

Article Title: IL-33 enhances proliferation and invasiveness of decidual stromal cells by up-regulation of CCL2/CCR2 via NF-κB and ERK1/2 signaling.

doi: 10.1093/molehr/gat094

Figure Lengend Snippet: Figure 8 Schematic representation of IL-33 effects on regulating biological behavior of DSCs. IL-33 can stimulate CCL2/CCR2 expression through NF-kB and ERK1/2 signal pathways, thereby enhancing proliferation and invasion of DSCs. At the same time, proliferation relative gene (PCNA, survivin) and invasion relative gene (titin, MMP2) are also increased in response to IL-33, which further promotes prolifertation and invasiveness of DSCs. sST2 can block IL-33/ST2 signaling through bounding to IL-33.

Article Snippet: BrdU cell proliferation assay Bromodeoxyuridine (5-bromo-2′-deoxyuridine, BrdU) cell proliferation assay was applied to evaluate the effect of IL-33 on cell proliferation with or without sST2, anti-CCL2 neutralizing antibody (MAB279, R&D Systems) and CCR2 blocker RS102895 (R1903, Sigma-Aldrich, Santa Clara, USA).

Techniques: Expressing, Blocking Assay

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Whole-lung murine IL-13, CCL6, and CCL21 levels in C.B-17SCID/bg mice that received normal, NSIP, or IPF/UIP fibroblasts. Lung samples were removed at days 35 and 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice, and all soluble proteins were measured by specific ELISA. Data shown are mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 compared with appropriate the C.B-17SCID/bg group that received normal fibroblasts. ττP ≤ 0.01, τττP ≤ 0.001 compared with whole-lung cytokine and chemokine levels at the day 35 time point in the C.B-17SCID/bg groups with either IPF/UIP or NSIP fibroblasts The whole-lung cytokine and chemokine levels in control C.B-17SCID/bg group that did not receive fibroblasts were as follows: IL-13, 0.18 ± 0.014 ng/mg protein; CCL6, 0.37 ± 0.05 ng/mg protein; and CCL21, 2.0 ± 0.1 ng/mg protein.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: SuperArray Analysis of Human CCR7 and CCL21 and TaqMan Analysis of Human CCR7 in Whole-Lung Samples from C.B-17SCID/bg Mice at Day 35 after i.v. Human Fibroblast Injection

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Injection

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Representative Mason trichrome-stained histological sections from C.B-17SCID/bg mice that received normal (A–C), NSIP (D–F), or IPF/UIP (G–I) fibroblasts. No interstitial remodeling was apparent in C.B-17SCID/bg mice that received normal fibroblasts, but vascular anomalies were observed in this group (B), and the IgG (A), anti-CCL21 monoclonal antibody (B), and anti-CCR7 monoclonal antibody (C) therapies did not alter the lung histological appearance in this group. Pulmonary remodeling was apparent in C.B-17SCID/bg mice that received NSIP fibroblasts, and this pattern was not altered by IgG (D), whereas the anti-CCL21 antibody (E) or anti-CCR7 antibody (F) therapies markedly reduced the interstitial remodeling in whole-lung samples. Interstitial pulmonary fibrosis was apparent in C.B-17SCID/bg mice that received IPF/UIP fibroblasts, and this pattern was not altered by IgG (G), whereas the anti-CCL21 antibody (H) or anti-CCR7 antibody (I) therapies markedly reduced the interstitial remodeling in whole-lung samples. Monoclonal antibody therapies began at day 35 and continued to day 63, and lung samples were removed at day 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice. Original magnification, ×400.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Staining

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Quantitative TaqMan PCR analysis of extracellular matrix-associated genes MMP-2 (top) and MMP-19 (bottom) in C.B-17SCID/bg mice that received normal, NSIP, or IPF/UIP fibroblasts. Changes in gene expression are expressed as mean ± SEM of the fold increase in transcript expression above a group of C.B-17SCID/bg mice that received PBS, PKH26, and one of IgG, anti-CCL21 antibody, and anti-CCR7 antibody. *P ≤ 0.05, ***P ≤ 0.001 compared with the appropriate C.B-17SCID/bg group that received human fibroblasts and IgG treatment.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Gene Expression, Expressing

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Whole-lung hydroxyproline levels in C.B-17SCID/bg mice that received no fibroblasts (ie, control) or received normal, NSIP, or IPF/UIP fibroblasts. All groups of mice received either IgG or monoclonal antibody therapy. IgG, anti-CCL21, and anti-CCR7 monoclonal antibody therapies began in separate groups of C.B-17SCID/bg mice at day 35 and continued to day 63. Lung samples were removed at day 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice. Data shown are mean ± SEM. *P ≤ 0.05 compared with the control C.B-17SCID/bg group, which did not receive any human fibroblasts; τττP ≤ 0.001 compared with the appropriate C.B-17SCID/bg group that received human fibroblasts and IgG treatment; τP ≤ 0.05 compared with the appropriate C.B-17SCID/bg group that received human fibroblasts and IgC treatment.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Control

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Whole-lung murine IL-13, CCL6, and CCL21 levels in C.B-17SCID/bg mice that received normal, NSIP, or IPF/UIP fibroblasts and IgG or monoclonal antibody therapies. Anti-CCL21 and anti-CCR7 monoclonal antibody therapies began at day 35 and continued to day 63. Lung samples were removed at day 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice. Specific ELISA was used to measure all soluble proteins. Data shown are mean ± SEM. *P ≤ 0.05, **P ≤ 0.01 compared with indicated protein levels measured in whole-lung samples from C.B-17SCID/bg mice that received human fibroblasts.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Enzyme-linked Immunosorbent Assay