Journal: BMC Biotechnology

Article Title: Active immunization with Tocilizumab mimotopes induces specific immune responses

doi: 10.1186/s12896-015-0161-9

Figure Lengend Snippet: Alignment of the insert sequences expressed by phage clones. Phe229, Tyr230, Glu278 and Phe279 from the D3 domain of site I contribute the majority to form the initial IL-6/IL-6Rα binary complex. We aligned the mimotope sequences and found that phage mimics 4A124, 4A125 and 4A126 bear these key amino acids (4A124: Phe229, Tyr230; 4A125: Glu278, Phe279; 4A126: Glu278, Phe279)

Article Snippet: Two days later, the medium was replaced by FBS-free medium with sera from mimotopes immunized mice with a dilution 1:100 or Tocilizumab (10 μg/ml, Roche Co. Ltd, Shanghai, China) at 37°C for 2 h. Then the cells were incubated with human IL-6 (40 ng/ml, R&D Systems, Minneapolis, MN) for 15 min at 37°C.

Techniques: Clone Assay

Journal: BMC Biotechnology

Article Title: Active immunization with Tocilizumab mimotopes induces specific immune responses

doi: 10.1186/s12896-015-0161-9

Figure Lengend Snippet: Signaling pathway analysis with mimotope-induced antibodies. We detected the levels of phosphorylated-STAT3, STAT3, phosphorylated-Erk1/2 and Erk1/2 in HeLa, Jurkat and U937 cells to observe whether IL-6 pathway could be down-regulated by when added sera from mice immunized with mimotopes. Cells were grown at 37 °C with medium containing 10 % FBS. Two days later, the medium was replaced by FBS-free medium with sera from mimotopes immunized mice with a dilution 1:100 or Tocilizumab (10 μg/ml) at 37 °C for 2 h. Then the cells were incubated with human IL-6 (40 ng/ml) for 15 min at 37 °C. In HeLa cells ( a ), incubation with sera from mice immunized with 4A125 and 4A126 could partly reduce the levels of phosphorylated-STAT3 and phosphorylated-Erk1/2, as compared to the control group. In Jurkat cells ( b ), incubation with sera from mice immunized with 4A220 and 4A126-KLH conjugate could reduce the level of STAT3, phosphorylated-Erk1/2 and Erk1/2. In U937 cells ( c ), incubation with sera from mice immunized with 4A124, 4a125 and 4A126-KLH conjugate could increase the level of phosphorylated- STAT3, STAT3 and phosphorylated-Erk1/2

Article Snippet: Two days later, the medium was replaced by FBS-free medium with sera from mimotopes immunized mice with a dilution 1:100 or Tocilizumab (10 μg/ml, Roche Co. Ltd, Shanghai, China) at 37°C for 2 h. Then the cells were incubated with human IL-6 (40 ng/ml, R&D Systems, Minneapolis, MN) for 15 min at 37°C.

Techniques: Incubation