title: In situ activated intestinal T cells expanded in vitro--without addition of antigen--produce IFN-gamma and IL-10 and preserve their function during growth.
abstract: OBJECTIVE
  The balance between mucosal CD4+ T cells producing IFN-gamma or IL-10 is essential in the maintenance of intestinal homeostasis. We aimed to investigate how in situ activated T cells were expanded in vitro according to a new cell culture protocol and if it selected for continuous clonal CD4+ T cells capable of producing mainly IFN-gamma or IL-10.
  
  
  METHODS
  T cell cultures were established from colonic biopsy specimens of 27 patients with Crohn's disease and from 10 healthy controls in a medium supplemented with IL-2 and IL-4 but without addition of exogenous antigen or mitogen. Cytokine production was measured after stimulation (IL-12, super antigen) and inhibition (cyclosporin and methylprednisolone).
  
  
  RESULTS
  Cytokine production was increased in cultures from patients with Crohn's disease compared to controls (IFN-gamma, p = 0.005; IL-10, p = 0.003; TNF-alpha, p = 0.03). Early cultures were highly responsive to IL-12 stimulation. We established CD4+ T-cell clones escaping cellular senescence with an inflammatory or regulatory cytokine profile.
  
  
  CONCLUSIONS
  The data indicate that cultures of in situ activated inflammatory and regulatory subpopulations of intestinal T lymphocytes with pathogenic importance in Crohn's disease can be established preserving their functional properties during growth.
authors.affiliation: Department of Medicine V, Aarhus University Hospital, Denmark. agnholt@dadlnet.dk
authors.forename: J
authors.initials: J
authors.lastname: Agnholt
authors.forename: K
authors.initials: K
authors.lastname: Kaltoft
mesh_terms: D000293:Adolescent; D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D016022:Case-Control Studies; D002999:Clone Cells; D003424:Crohn Disease / Q000276:immunology*; D016572:Cyclosporine / Q000494:pharmacology; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000096:biosynthesis*; D016753:Interleukin-10 / Q000096:biosynthesis*; D018664:Interleukin-12 / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D008775:Methylprednisolone / Q000494:pharmacology; D008875:Middle Aged; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D018664:Interleukin-12; D007371:Interferon-gamma; D016572:Cyclosporine; D008775:Methylprednisolone