NON-NEURONAL CHOLINERGIC SYSTEM MODULATE THE CROSS-TALK BETWEEN THE IMMUNE SYSTEM AND GLIOBLASTOMA CELLS

VANINA FONTANA, ; ANTONELA ASAD ; MARIANELA CANDOLFI; GABRIELA SALAMONE,

Congreso; Reunión conjunta de Sociedades de Biociencia; 2017

Glioblastoma multiforme (GBM) is the deadliest and most common type of human primary brain tumor. This tumor is defined by the hallmark features of uncontrolled cellular proliferation, diffuse infiltration, robust angiogenesis, resistance to apoptosis and genomic instability. Acetylcholine is a neurotransmitter which can also modulates cell survival, proliferation and differentiation in neuronal and non-neuronal cells such as immune cells, which has been referred to as a ?non-neuronal cholinergic system?. The aim of this work was to elucidate the relevance of the non-neuronal cholinergic system in the interaction between immune and GBM cells. We first evaluated the expression of acetylcholine receptors in human GBM cell lines by fluorescence microscopy. We found that both U251 and U373 human GBM cells express acetylcholine muscarinic receptors M1 and M3. In order to evaluate whether the cholinergic system affects the cross-talk with immune cells, human U251 cells were co-cultured with human dendritic cells (DC) in the presence of cholinergic agonists (carbachol and muscarine). Mononuclear cells were isolated from buffy coats of healthy adult nonsmoker volunteer and CD14+ cells were then isolated by positive selection and then were cultured with GM-CSF and IL-4. The co-cultures were incubated in the presence of carbachol 10-8 M) and muscarine (10-8M). We found that U251 cells upregulated the expression of CD86 in DCs as assessed by flow cytometry in presence of carbachol and muscarine with respect to control co-cultures (p