Identification of Cells Responding to Vasoactive Intestinal Peptide by Measuring Intracellular Cyclic Adenosine Monophosphate in Human Colonic Mucosa

Abstract

Background: Although there is great deal of evidence suggesting that vasoactive intestinal peptide (VIP) has immunomodulating effects on human colonic lamina propria mononuclear cells (LPMC), it remains unclear which type of cell carries functional VIP receptors. In this study we investigated the presence of functional VIP receptors by measuring intracellular cyclic adenosine monophosphate (cAMP) in isolated epithelial cells, bulk LPMC, T cells, and macrophages in human colonic mucosa. Methods: Epithelial cells and LPMC were isolated from non-pathologic segment of colonic mucosa of surgical specimens from five patients with colonic cancer. Mucosal T cells and macrophages were further isolated from LPMC. Each cell population was cultured with various concentration of VIP for 60 min at most. Then, intracellular cAMP was extracted and measured by enzyme-linked immunoassay. Results: When isolated epithelial cells were examined, VIP increased intracellular cAMP in a dose-dependent fashion, as observed in HT-29 cells used as a positive control. In contrast, the concentration of cAMP was essentially stable in response to VIP when isolated LPMC were examined. This was the case even when separated T cells and macrophages were individually investigated. To evaluate the possible effects of enzyme digestion for LPMC isolation on the VIP response, HT-29 cells were precultured with collagenase and deoxyribonuclease (DNase I), resulting in less enhancement of cAMP by VIP. Conclusions: In this study we failed to show VIP-responsive enhancement of cAMP in mucosal immune cells, suggesting that epithelial cells may be major effecter cells of VIP in human colonic mucosa.