title: Generation of highly purified and functionally active human TH1 cells against Aspergillus fumigatus.
abstract: Invasive aspergillosis remains a serious complication in patients undergoing allogeneic stem cell transplantation (SCT). Since it became clear that lymphocytes provide a critical secondary defense against fungi, adoptive transfer of functionally active anti-Aspergillus T cells might be an option to restore adaptive immune effector mechanisms. Using the interferon (IFN)-gamma secretion assay, we isolated human activated T cells upon stimulation with a cellular extract of Aspergillus fumigatus. Culturing this cell population for 14 days, we obtained an average of 1.1 x 10(7) cells from a single 100-mL blood draw in 7 of 7 healthy individuals. Within another 14 days, these cells were expanded to an average number of 2.0 x 10(8) T-helper 1 (T(H)1) cells secreting IFN-gamma on stimulation with Aspergillus antigens. Testing various fungal antigen extracts, similar proportions of IFN-gamma-producing CD3+/CD4+ cells were obtained upon activation with antigen extracts of A. fumigatus, A. flavus, A. niger, and Penicillium chrysogenum, whereas no significant IFN-gamma production was observed upon activation with antigen extracts of Alternaria alternata and Candida albicans. In addition, generated T cells were able to induce damage to A. fumigatus hyphae, and significantly increased hyphal damage induced by human neutrophils. CD4+ T-cell-mediated alloreactivity of generated anti-Aspergillus T cells was clearly reduced compared with that of the original cell population. In conclusion, we present a simple and feasible strategy for rapid generation of a high number of functional active T cells against Aspergillus from a single blood draw. Our data suggest that functionally active T cells against Aspergillus could be a promising treatment option for patients undergoing allogeneic SCT.
authors.affiliation: Pediatric Hematology and Oncology, Johann Wolfgang Goethe University, Theodor-Stern-Kai 7, D-60590 Frankfurt, Germany.
authors.forename: Olaf
authors.initials: O
authors.lastname: Beck
authors.forename: Max S
authors.initials: MS
authors.lastname: Topp
authors.forename: Ulrike
authors.initials: U
authors.lastname: Koehl
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Roilides
authors.forename: Maria
authors.initials: M
authors.lastname: Simitsopoulou
authors.forename: Mitra
authors.initials: M
authors.lastname: Hanisch
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Sarfati
authors.forename: Jean Paul
authors.initials: JP
authors.lastname: Latgé
authors.forename: Thomas
authors.initials: T
authors.lastname: Klingebiel
authors.forename: Hermann
authors.initials: H
authors.lastname: Einsele
authors.forename: Thomas
authors.initials: T
authors.lastname: Lehrnbecher
mesh_terms: D019264:Adoptive Transfer / Q000379:methods; D000946:Antigens, Fungal / Q000276:immunology / Q000494:pharmacology; D001228:Aspergillosis / Q000517:prevention & control*; D001232:Aspergillus fumigatus / Q000276:immunology*; D018929:Cell Culture Techniques; D006801:Humans; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology*
chemical_list: D000946:Antigens, Fungal; D007371:Interferon-gamma