Foxp3- IgG(Fc): Capable in inducing humoral immune responses against regulatory T-cells
Subject Areas : biotechnology
maryam naseroleslami
1
,
neda mousaviniri
2
1 - Department of Cellular and Molecular Biology, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran
2 - Department of Medical Biotechnology, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran
Keywords: FoxP3, Fc, Immunotherapy, Humoral response, T regulatory,
Abstract :
It seems that the success of vaccination in order to cancer immunotherapy is abrogating through a powerful network of immune system suppressive elements which one of them is regulatory T cell. In different studies inducing an immune response against regulatory cells expressing foxp3 and their removal have been assessed. As these previous studies could not efficiently conquer the suppressive effect of regulatory cells by their partial elimination, we were searching for constructing more effective vaccines against regulatory T cells by which we hope to improve the effect of combined means of immunotherapy in cancer. DNA construct containing fragment C (Fc) portion of IgG fused to Foxp3 was designed. Expression of DNA construct was investigated following its transfection into HEK cells through fluorescent microscopy, flow cytometry and western blot. FOXP3-Fc fusion protein was expressed in E. coli strain BL21 as host cells. DNA construct and respective protein were injected into C57BL/6 mice. After 2 weeks, serum level of IgG antibody and its sub classes was investigated by ELISA test. The expression analysis of DNA construct showed that this construct successfully expressed in eukaryotic cells. Moreover, the Foxp3-Fc expression was confirmed by SDS-PAGE followed by western blot analysis.
_||_
