Fibroblast and T cells conditioned media induce maturation dendritic cell and promote T helper immune response

Masoumeh Asadi, Farah Farokhi, Nowruz Delirezh,Meysam Ganji Bakhsh,  Vahid Nejati, Keykavos Golami

Abstract
Dendritic cells (DCs) induce pathogen-specific T cell responses. We comprehensively studied the effects of addition of maturation stimulus, fibroblasts (fibroblast conditioned medium), PHA activated T cells (T cell conditioned medium), and mixture of fibroblast & PHA activated T cells (FCM-TCCM) conditioned media on maturation of DCs. Monocytes were cultured with GM-CSF and IL-4 for five days. Maturation factors included MCM and TNF-α as control group. FCM and TCCM, or FCM-TCCM supernatant were considered as the treatment group. Tumor antigens were added at day five. Matured DCs were harvested at day seven. Phenotypic and functional analyses were carried out using anti (CD14, CD80, CD86, CD83 and HLA-DR) monoclonal antibodies. Phagocytic activity, mixed lymphocyte reaction (MLR) and cytokine production were also evaluated. At the end of culturing period, significantly fully matured DCs with large amount cytoplasm and copious dendritic projections were found in the presence of MCM, TNF-α with or without FCM, TCCM, FCM as well as TCCM. Flow cytometric analysis revealed that expression of CD14 decreased in particular in treated DCs, at the 5th day and expression of CD80, CD86 and HLA-DR was higher when FCM, TCCM, FCM plus TCCM were added to maturation factor. This study demonstrated that DCs matured with these methods had optimum function in comparison with either factor alone.

Key words: Dendritic cell, Fibroblast, T cell