Journal: Cancer Research

Article Title: The CXCR5 Chemokine Receptor Is Expressed by Carcinoma Cells and Promotes Growth of Colon Carcinoma in the Liver

doi: 10.1158/0008-5472.can-06-1507

Figure Lengend Snippet: Figure 2. FACS analysis of CXCR5 and CXCR4 surface levels on mouse and human carcinoma cell lines. A, MCA38 mouse colon and MB49 mouse bladder carcinoma in vitro and in vivo (isolated from s.c. tumors). B, pancreatic carcinoma, human CAPAN-1 and CAPAN-2 in vitro, and mouse Panc02 in vitro and in vivo. C, MDA-MB-231 human mammary carcinoma in vitro and in vivo. D, a CXCR4+ variant of the MDA-MB-231 carcinoma, isolated from a minor subpopulation, in vitro and in vivo.

Article Snippet: Cultures were trypsinized and 106 cells were incubated for 60 minutes with phycoerythrin-conjugated CXCR4 antibody (12G5, R&D Systems, Minneapolis, MN) or with CXCR5 antibodies (clone 51505.111, R&D systems) and then phycoerythrin-conjugated antibodies against mouse IgG and analyzed using a Becton Dickinson (Franklin Lakes, NJ) FACSCalibur using CellQuest software.

Techniques: In Vitro, In Vivo, Isolation, Variant Assay

Journal: Cancer Research

Article Title: The CXCR5 Chemokine Receptor Is Expressed by Carcinoma Cells and Promotes Growth of Colon Carcinoma in the Liver

doi: 10.1158/0008-5472.can-06-1507

Figure Lengend Snippet: Figure 1. A, RT-PCR assay of CXCL13 expression in different mouse organs, with h-actin as control. B-D, FACS analysis of CXCR5 surface levels on CT26 colon carcinoma cells. B, cells in vitro and isolated from a spleen tumor, liver metastases, and a s.c. tumor. C, cells isolated from lungs, 2 and 7 days after tail vein injection of 2 106 cells. D, cells isolated from lung metastases and the same cells after 1 and 4 days in vitro.

Article Snippet: Cultures were trypsinized and 106 cells were incubated for 60 minutes with phycoerythrin-conjugated CXCR4 antibody (12G5, R&D Systems, Minneapolis, MN) or with CXCR5 antibodies (clone 51505.111, R&D systems) and then phycoerythrin-conjugated antibodies against mouse IgG and analyzed using a Becton Dickinson (Franklin Lakes, NJ) FACSCalibur using CellQuest software.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control, In Vitro, Isolation, Injection

Journal: Cancer Research

Article Title: The CXCR5 Chemokine Receptor Is Expressed by Carcinoma Cells and Promotes Growth of Colon Carcinoma in the Liver

doi: 10.1158/0008-5472.can-06-1507

Figure Lengend Snippet: Figure 3. CXCR5 expression in human pancreatic ductal carcinoma. Strong cytoplasmic expression in a moderately (A) and in poorly differentiated carcinomas (B-C) in tumor cells (T) and in scattered inflammatory cells (arrows). D, reactive lymph node that served as positive control. B lymphocytes in germinal center (GC) and mantel zone are moderately to strongly stained. Immunoperoxidase staining, 200 magnification.

Article Snippet: Cultures were trypsinized and 106 cells were incubated for 60 minutes with phycoerythrin-conjugated CXCR4 antibody (12G5, R&D Systems, Minneapolis, MN) or with CXCR5 antibodies (clone 51505.111, R&D systems) and then phycoerythrin-conjugated antibodies against mouse IgG and analyzed using a Becton Dickinson (Franklin Lakes, NJ) FACSCalibur using CellQuest software.

Techniques: Expressing, Positive Control, Staining, Immunoperoxidase Staining

Journal: Cancer Research

Article Title: The CXCR5 Chemokine Receptor Is Expressed by Carcinoma Cells and Promotes Growth of Colon Carcinoma in the Liver

doi: 10.1158/0008-5472.can-06-1507

Figure Lengend Snippet: Figure 4. Properties of CXCR5-transfected CT26 cells in vitro. A, chemotaxis assay, with or without pertussis toxin pretreatment. Values are number of cells per field on the lower surface of a filter in a Transwell with or without 100 ng/mL CXCL13 in the lower well. B, proliferation assay of control CT26 and CXCR5- CT26 cells in the presence or absence of 100 ng/mL CXCL13, in either keratinocyte medium (kerat. med.) + 1% FCS (medium with minimal control proliferation) or regular DMEM + 10% FCS. Hexosaminidase activity was measured and converted to cell number as calculated from a standard curve.

Article Snippet: Cultures were trypsinized and 106 cells were incubated for 60 minutes with phycoerythrin-conjugated CXCR4 antibody (12G5, R&D Systems, Minneapolis, MN) or with CXCR5 antibodies (clone 51505.111, R&D systems) and then phycoerythrin-conjugated antibodies against mouse IgG and analyzed using a Becton Dickinson (Franklin Lakes, NJ) FACSCalibur using CellQuest software.

Techniques: Transfection, In Vitro, Chemotaxis Assay, Proliferation Assay, Control, Activity Assay

Journal: Cancer Research

Article Title: The CXCR5 Chemokine Receptor Is Expressed by Carcinoma Cells and Promotes Growth of Colon Carcinoma in the Liver

doi: 10.1158/0008-5472.can-06-1507

Figure Lengend Snippet: Figure 5. CT26 cells transfected with CXCR5 or the CXCL13-KDEL ‘‘intrakine’’ in vivo. A, CXCR5 surface levels in spleen tumors: almost no CXCR5 on surface of intrakine-transfected cells. Results of RT-PCR with primers specific for mouse CXCR5 that reveal induction of CXCR5 mRNA in vivo in all CT26 cells and Western blot results showing that CXCR5 protein is present in the CXCL13- KDEL-CT26 cells to a similar extent as in the other CT26 cells. Note that the CXCR5 antibody reacts with the transfected human CXCR5 as well as the endogenous mouse CXCR5. B, liver weight of mice, 14 days after injection of 106 cells into the spleen. Normal liver weight (f2 g). Bars, SD. Ps were obtained by t tests comparing CXCR5-CT26 and CXCL13-KDEL-CT26 cells, respectively, with control cells. C, similar as in (B), but for the spleen weights. D, similar as in (B), but for tumors generated by s.c. injection.

Article Snippet: Cultures were trypsinized and 106 cells were incubated for 60 minutes with phycoerythrin-conjugated CXCR4 antibody (12G5, R&D Systems, Minneapolis, MN) or with CXCR5 antibodies (clone 51505.111, R&D systems) and then phycoerythrin-conjugated antibodies against mouse IgG and analyzed using a Becton Dickinson (Franklin Lakes, NJ) FACSCalibur using CellQuest software.

Techniques: Transfection, In Vivo, Reverse Transcription Polymerase Chain Reaction, Western Blot, Injection, Control, Generated

Journal: Cancer Research

Article Title: The CXCR5 Chemokine Receptor Is Expressed by Carcinoma Cells and Promotes Growth of Colon Carcinoma in the Liver

doi: 10.1158/0008-5472.can-06-1507

Figure Lengend Snippet: Figure 6. Bioluminescence of liver metastases, generated by intrasplenic injection of 5 104 CT26 cells expressing luciferase (CT26-luc), transfected with CXCR5 or CXCL13-KDEL or nontransfected controls. The spleen was removed after injection. Of two separate experiments, only the mice are shown that did not develop a tumor at the injection site. In total, three or four mice are shown for each group. Note logarithmic scale.

Article Snippet: Cultures were trypsinized and 106 cells were incubated for 60 minutes with phycoerythrin-conjugated CXCR4 antibody (12G5, R&D Systems, Minneapolis, MN) or with CXCR5 antibodies (clone 51505.111, R&D systems) and then phycoerythrin-conjugated antibodies against mouse IgG and analyzed using a Becton Dickinson (Franklin Lakes, NJ) FACSCalibur using CellQuest software.

Techniques: Generated, Injection, Expressing, Luciferase, Transfection