title: Human gammadelta T cells from G-CSF-mobilized donors retain strong tumoricidal activity and produce immunomodulatory cytokines after clinical-scale isolation.
abstract: Human gammadelta T cells are a small fraction of T cells that have been shown to exert major histocompatibility (MHC)-unrestricted natural cytotoxicity against a variety of solid tumors and some subsets of leukemias and lymphomas. They are also involved in the immune response to certain bacterial, viral, and parasitic infections and expand significantly in CMV- or HSV-infected organ allografts. They are able to mediate antibody-dependent cytotoxicity and are not alloreactive, which makes them attractive candidates for cell-based immunotherapy. However, their frequency in peripheral blood is low and ex vivo expansion of gammadelta T cells is labor-extensive, does not always yield cells with full innate cytotoxic power, and has the potential for microbial contamination. Therefore, the authors developed a clinical-scale, automated cell purification method for the efficient enrichment of gammadelta T cells from leukapheresis products. Six leukapheresis products were purified for gammadelta T cells using a single-step immunomagnetic method. Purity and phenotype were assessed by flow cytometry. A standard Europium release assay was performed to determine the cytotoxic capacity of the cells. Cytokine production was measured using a multiplex sandwich immunoassay. The mean percentage of gammadelta T cells in the final product was 91%, with an average recovery of 63%. The cells showed a high co-expression of CD8, CD56, CD28, and CD11b/CD18. In some products an unusually high proportion of Vgamma9Vdelta1 T cells was found. The isolated cells were cytotoxic against the neuroblastoma cell line NB1691 and the erythroleukemic line K562 in vitro. They were able to produce a variety of immunomodulatory cytokines such as IFNgamma, TNFalpha, and MIP-1beta, but also GM-CSF and G-CSF when co-incubated in culture with and without various stimuli. In summary, the authors describe a rapid, automated, and efficient method for the large-scale enrichment of human gammadelta T cells. The cytotoxic properties of the cells were preserved. This method yields sufficient purified gammadelta T cells for use in adoptive immunotherapy as well as laboratory investigations and animal studies.
authors.affiliation: Department of Hematology-Oncology, Mail-Stop 321, St. Jude Children's Research Hospital, 332 N. Lauderdale Street, Memphis, TN 38105-2794, USA. mario.otto@stjude.org
authors.forename: Mario
authors.initials: M
authors.lastname: Otto
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Barfield
authors.forename: Rekha
authors.initials: R
authors.lastname: Iyengar
authors.forename: Janet
authors.initials: J
authors.lastname: Gatewood
authors.forename: Ingo
authors.initials: I
authors.lastname: Müller
authors.forename: Martha S
authors.initials: MS
authors.lastname: Holladay
authors.forename: Jim
authors.initials: J
authors.lastname: Houston
authors.forename: Wing
authors.initials: W
authors.lastname: Leung
authors.forename: Rupert
authors.initials: R
authors.lastname: Handgretinger
mesh_terms: D001782:Blood Donors; D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism*; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods; D007937:Leukapheresis; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism
chemical_list: D016207:Cytokines; D016692:Receptors, Antigen, T-Cell, gamma-delta; D016179:Granulocyte Colony-Stimulating Factor