rat anti mouse mab 470  (R&D Systems)

Journal: BMC Immunology

Article Title: CXCL13 antibody for the treatment of autoimmune disorders

doi: 10.1186/s12865-015-0068-1

Figure Lengend Snippet: Effect on CXCL13- (A-D) and CXCL12- (negative control; E) induced migration of human and murine cells. Human pre-B-697-hCXCR5 cells (A, E) , human pre-B-697-hCXCR4 cells (E) , human tonsillar cells (B, E) , C57BL/6 splenocytes (C, E) , and SJL mouse splenocytes (D, E) were seeded into Transwell inserts. Diluted chemokines +/− antibodies were added into the lower chambers and plates were incubated for 2 h at 37°C (50 ug/ml of each antibody were used in CXCL12-induced migration assay). Inserts were removed and Alamar blue was added to the wells and incubated at 37°C overnight. Fluorescence was measured at wavelengths of 530 nm and 590 nm and the percent inhibition of chemokine-induced migration (A-D) or migration index (E) was calculated. Data represents an average of the measurements from at least three independent experiments + SEM. Statistical significance (relative to the isotype controls) was evaluated using one-way ANOVA followed by Bonferroni’s multiple comparison post test **** P < 0.0001; *** P < 0.0003; **P < 0.01.

Article Snippet: In an attempt to decipher the mechanism of action of anti-CXCL13 antibody in animal models, Finch et al. [ ] examined the integrity of splenic architecture in BALB/c mice immunized with alum-precipitated DNP-OVA and treated with rat anti-mouse CXCL13 antibody MAb 470 (R&D Systems, clone 143614).

Techniques: Negative Control, Migration, Incubation, Fluorescence, Inhibition, Comparison

Journal: BMC Immunology

Article Title: CXCL13 antibody for the treatment of autoimmune disorders

doi: 10.1186/s12865-015-0068-1

Figure Lengend Snippet: Inhibition of human CXCL13-mediated internalization of human CXCR5 receptor. Human pre-B-697-huCXCR5 cells were pre-blocked with anti-human Fc for 15 min at 37°C, incubated with a pre-complexed huCXCL13/antibody mix for 2 h at 37°C, and subsequently stained with anti-human CXCR5 antibody for flow cytometric analysis. (A) Cell surface CXCR5 receptor expression. Geometric mean values were determined by FlowJo 7.6 software. (B) EC50 values were calculated from sigmoidal dose response curves (shown on the graph) with R 2 values equal to 0.9 (MAb 5261) and 0.98 (MAb 5261-muIg). Data points represent an average of measurements obtained from 3 independent experiments.

Article Snippet: In an attempt to decipher the mechanism of action of anti-CXCL13 antibody in animal models, Finch et al. [ ] examined the integrity of splenic architecture in BALB/c mice immunized with alum-precipitated DNP-OVA and treated with rat anti-mouse CXCL13 antibody MAb 470 (R&D Systems, clone 143614).

Techniques: Inhibition, Incubation, Staining, Expressing, Software

Journal: BMC Immunology

Article Title: CXCL13 antibody for the treatment of autoimmune disorders

doi: 10.1186/s12865-015-0068-1

Figure Lengend Snippet: Effect of murine anti-CXCL13 antibody on germinal center formation in C57BL/6 mice challenged with NP-KLH. Mice were treated with 30 mg/kg of MAb 5261-muIg antibody and corresponding mouse isotype control, on days −3, 0 (the day of the immunization), 4 and 7 (n = 9/group). (A) Total IgM and IgG1 anti-NP antibody-secreting cells were detected by ELISPOT on wells coated with NP14-BSA. (B) The frequencies of high affinity IgG1 anti-NP-secreting cells were determined by ELISPOT on wells coated with NP4-BSA. (C) Number of germinal centers (n = 9 mice/group). Follicles were hand counted. Number of PNA+ germinal centers in relation to total number of follicles per group was calculated using ImagePro software. (D) Mean germinal center area. Area of each PNA+ germinal center for each group was calculated using ImagePro software by PNA+ pixel area and then converted to μm. The numbers of GC analyzed were: 201 in Control group and 102 in MAb 5261-muIg-treated group. (E) Percentages of activated T cells (CD4+/CD62Llow/CD44+) in spleens and bone marrow. Statistical significance was evaluated by Student’s unpaired t -test. *P = 0.0125; **P = 0.0016; ****P < 0.0001; P = 0.2 and 0.71 for IgG1 and IgM portions of the graph, respectively (A) .

Article Snippet: In an attempt to decipher the mechanism of action of anti-CXCL13 antibody in animal models, Finch et al. [ ] examined the integrity of splenic architecture in BALB/c mice immunized with alum-precipitated DNP-OVA and treated with rat anti-mouse CXCL13 antibody MAb 470 (R&D Systems, clone 143614).

Techniques: Control, Enzyme-linked Immunospot, Software