Successful in vitro immortalization of human intestinal mucosal lymphocytes with Herpesvirus saimiri

Mucosal intestinal lymphocytes form the first immune-cell line of defense i n the intestine. Several methodologies, most of them cumbersome and time co nsuming, have been used to obtain T-cell clones to unveil their physiologic al role. In the present work We take advantage of the recently described te chnique of transformation of T lymphocytes using Herpesvirus saimiri to sho w that it is possible to immortalize intestinal T-cell lines derived from h ealthy and diseased colonic samples and thence easily obtain in vitro intes tinal T-cell lines as a model for physiopathological studies. Intestinal sa mples were obtained by colonoscopy and digested with dispase and collagenas e. Mucosal lymphocytes (assessed by the expression of the CD3 and CD103 mar kers) were isolated using a Percoll gradient centrifugation and transformed with Herpesvirus saimiri Sustained growth was observed 3 months later, sho wing that the cells were successfully transformed, a finding further confir med by PCR. All cell lines were CD8(+)TcR alpha beta(+) and HLA-DR+. CD25 w as expressed on 1% of Crohn's disease-derived cells and on 25% of cells der ived from patients with ulcerative colitis. CD80 expression was found on 89 -90% of the cells. These immortal cell lines of intestinal origin may be us eful in future experiments aimed at elucidating the role of mucosal lymphoc ytes in health and disease.