Journal: Journal of Virology

Article Title: In Vitro Neutralization Is Not Predictive of Prophylactic Efficacy of Broadly Neutralizing Monoclonal Antibodies CR6261 and CR9114 against Lethal H2 Influenza Virus Challenge in Mice

doi: 10.1128/JVI.01603-17

Figure Lengend Snippet: Evaluation of CR6261 and CR9114 in neutralization assays against H2 influenza viruses

Article Snippet: A/Ann Arbor/6/1960 (H2N2) is a recombinant wild-type virus provided by MedImmune, Gaithersburg, MD.

Techniques: Neutralization, Recombinant

Journal: Journal of Virology

Article Title: In Vitro Neutralization Is Not Predictive of Prophylactic Efficacy of Broadly Neutralizing Monoclonal Antibodies CR6261 and CR9114 against Lethal H2 Influenza Virus Challenge in Mice

doi: 10.1128/JVI.01603-17

Figure Lengend Snippet: Prophylactic efficacy of high-dose CR6261 and CR9114 against H2 influenza virus challenge in BALB/c mice. To determine if bNAbs protected against H2 influenza virus challenge, BALB/c mice (n = 8/group) were administered CR6261, CR9114, or CR-JB (isotype control) at a dose of 15 mg/kg, or vehicle (antibody dilution buffer), and were challenged with 25 MLD50 of A/Ann Arbor/6/1960 (H2N2) or A/swine/MO/42964624/2006 (H2N3). (A and B) Percent survival (A) and percent weight loss (C) (mean ± standard error of the mean [SEM]) after A/Ann Arbor/6/1960 (H2N2) challenge. (B and D) Percent survival (B) and percent weight loss (D) (mean ± SEM) after A/swine/MO/42964624/2006 (H2N3) challenge. Daggers represent a vehicle-treated mouse that succumbed to infection or that reached endpoint.

Article Snippet: A/Ann Arbor/6/1960 (H2N2) is a recombinant wild-type virus provided by MedImmune, Gaithersburg, MD.

Techniques: Virus, Control, Infection

Journal: Journal of Virology

Article Title: In Vitro Neutralization Is Not Predictive of Prophylactic Efficacy of Broadly Neutralizing Monoclonal Antibodies CR6261 and CR9114 against Lethal H2 Influenza Virus Challenge in Mice

doi: 10.1128/JVI.01603-17

Figure Lengend Snippet: Prophylactic efficacy of decreasing doses of CR6261 and CR9114 against H2 influenza virus challenge in BALB/c mice. To evaluate efficacy over a range of antibody doses, BALB/c mice were administered CR6261 or CR9114 at doses of 0, 1.7, 5, and 15 mg/kg and were challenged with 25 MLD50 of H2 influenza viruses. (A and C) Percent survival (A) and percent weight loss (C) (mean ± SEM) after A/Ann Arbor/6/1960 (H2N2) challenge. (B and D) Percent survival (B) and percent weight loss (D) (mean ± SEM) after A/swine/MO/42964624/2006 (H2N3) challenge. Eight to 10 animals were used in each group, and animals were monitored for 14 days after virus challenge.

Article Snippet: A/Ann Arbor/6/1960 (H2N2) is a recombinant wild-type virus provided by MedImmune, Gaithersburg, MD.

Techniques: Virus

Journal: Journal of Virology

Article Title: In Vitro Neutralization Is Not Predictive of Prophylactic Efficacy of Broadly Neutralizing Monoclonal Antibodies CR6261 and CR9114 against Lethal H2 Influenza Virus Challenge in Mice

doi: 10.1128/JVI.01603-17

Figure Lengend Snippet: Pulmonary viral load following CR6261 and CR9114 prophylaxis and H2 influenza virus challenge in mice. (A and B) BALB/c mice (n = 4 per time point) were given CR6261 or CR9114 (5 mg/kg) and then challenged 24 h later with 25 MLD50 of either A/Ann Arbor/6/1960 (H2N2) (A) or A/swine/MO/42964624/2006 (H2N3) (B). A vehicle (0 mg/kg) control was also included and challenged. On days 2 and 4 p.i., lung tissue homogenates were prepared and titrated on MDCK cells. Black bars indicate mean titers, while symbols represent titers for individual animals. An asterisk denotes significant difference from the vehicle control (P < 0.05).

Article Snippet: A/Ann Arbor/6/1960 (H2N2) is a recombinant wild-type virus provided by MedImmune, Gaithersburg, MD.

Techniques: Virus, Control

Journal:

Article Title: Activin A in the Regulation of Corneal Neovascularization and Vascular Endothelial Growth Factor Expression

doi:

Figure Lengend Snippet: Activin A up-regulates VEGF expression in corneal epithelial cells in vitro. Cells were treated overnight with recombinant activin A or the inhibitors as described in the Materials and Methods section and VEGF protein levels were measured in their supernatant. Bars represent the VEGF protein levels released in the supernatant after the various treatments (mean ± SD). Treatment with activin A, but not with the vehicle (control) up-regulated VEGF protein levels, whereas inhibition of p42/44 MAPK or p38 MAPK suppressed activin A-induced VEGF up-regulation.

Article Snippet: Corneal lysates, or recombinant phopsho-p38 MAPK standards (provided by the manufacturer), were subsequently incubated with a monoclonal antibody against the phosphorylated (activated) form of p38 MAPK (Assay Designs), immobilized on a microtiter 96-well plate for 1 hour at room temperature on a plate shaker at 500 rpm.

Techniques: Expressing, In Vitro, Recombinant, Control, Inhibition

Journal:

Article Title: Activin A in the Regulation of Corneal Neovascularization and Vascular Endothelial Growth Factor Expression

doi:

Figure Lengend Snippet: Activin A up-regulates both p38 and p42/44 MAPK activity in corneal epithelial cells. Cells were treated with activin A and p38 (A) and p42/44 (B) MAPK activity was measured as described in the Materials and Methods section. Bars represent the 450-nm reading that corresponds to the p38 or p42/44 MAPK activity (mean ± SD). Treatment with activin A up-regulated both p38 and p42/44 MAPK activity.

Article Snippet: Corneal lysates, or recombinant phopsho-p38 MAPK standards (provided by the manufacturer), were subsequently incubated with a monoclonal antibody against the phosphorylated (activated) form of p38 MAPK (Assay Designs), immobilized on a microtiter 96-well plate for 1 hour at room temperature on a plate shaker at 500 rpm.

Techniques: Activity Assay

Journal:

Article Title: Activin A in the Regulation of Corneal Neovascularization and Vascular Endothelial Growth Factor Expression

doi:

Figure Lengend Snippet: P42/44 (A) and p38 (B) kinase activity increases during the course of corneal neovascularization in our murine model. Animals were treated as described in the Materials and Methods section, sacrificed on day 0 (D0) or day 7 (D7) after the scraping, and the activities of p42/44 and p38 were measured in corneal lysates. Bars represent the 450-nm reading that corresponds to the p42/44 (A) or p38 (B) kinase activity, respectively (mean ± SD). P42/44 and p38 kinase activity increased during corneal neovascularization in the murine model paralleling the increases in both activin A and VEGF. Administration of recombinant activin A (ActA) increased even further the activity of p42/44 and p38 MAPK, whereas administration of a neutralizing antibody against activin A (AntiA) decreased the activation of both p42/44 and p38 MAPK. Administration of the isotype-matched control antibody did not have a significant effect on the amount of either p38 or p42/44 MAPK.

Article Snippet: Corneal lysates, or recombinant phopsho-p38 MAPK standards (provided by the manufacturer), were subsequently incubated with a monoclonal antibody against the phosphorylated (activated) form of p38 MAPK (Assay Designs), immobilized on a microtiter 96-well plate for 1 hour at room temperature on a plate shaker at 500 rpm.

Techniques: Activity Assay, Recombinant, Activation Assay, Control