Fig. 5
From: Active immunization with Tocilizumab mimotopes induces specific immune responses
Mimotope-induced antibodies for IL-6R recognition. a Titers of the six mouse groups determined by ELISA (the fourth immune serum, day 79) against rIL-6R. b HeLa, Jurkat and U-937 cell lysates were loaded with 50 μg/per lane. IL-6R expressed in HeLa, Jurkat and U-937 detection using Tocilizumab (lane 1) and the sera from mice immunized with the 4A220-KLH conjugate (lane 2), the 4A124-KLH conjugate (lane 3), 4A125-KLH conjugate (lane 4), 4A126-KLH conjugate (lane 5), the control peptide-KLH conjugate (lane 6), or the carrier KLH alone (lane 7). Sera were diluted with 1:1000. The results showed that the antibodies in the sera of the mice immunized with mimotopes could recognize sIL-6R (55KDA) and/or mIL-6R (80KDA) expressed in HeLa, Jurkat and U-937 cell line. c For antibody competitive assays, HeLa cell lysates were loaded with 40 μg/per lane. IL-6R expressed in HeLa was detected by using sera from mice immunized with the 4A220-KLH conjugate (lane 1), Tocilizumab+ sera from mice immunized with the 4A220-KLH conjugate (lane 2), sera from mice immunized with the 4A124-KLH conjugate (lane 3), Tocilizumab+ sera from mice immunized with the 4A124-KLH conjugate (lane 2), sera from mice immunized with the 4A125-KLH conjugate (lane 5), Tocilizumab+ sera from mice immunized with the 4A125-KLH conjugate (lane 2), sera from mice immunized with the 4A126-KLH conjugate (lane 7) and Tocilizumab+ sera from mice immunized with the 4A220-KLH conjugate (lane 8). Sera were diluted with 1:1000. The results showed that antibodies induced by mimotopes-KLH conjugate could compete with Tocilizumab to IL-6R expressed in HeLa cells, except antibodies induced by 4A124-KLH conjugate