title: Long-term culture growth of CD4-CD8- lymphocytes exhibiting elevated non-MHC-restricted cytotoxic activity.
abstract: We have developed a culture system for "long-term" growth of human lymphokine-activated killer (LAK) cells exhibiting an elevated, wide-spectrum anti-tumor cytotoxicity. The system allows the exponential growth of monocyte- and B-lymphocyte-depleted CD4-CD8- lymphocytes in the presence of human AB serum and recombinant human interleukin-2 (IL-2) (2 x 10(2) U/ml) combined with interleukin (IL-1) beta (50 ng/ml). After 21 days in culture, these cells undergo massive amplification (i.e., the cell yield rises up to 30-120 times the starting values), and exhibit a marked anti-tumor cytotoxic activity against a panel of natural killer (NK)-resistant tumor cell lines. Interestingly, this activity correlates with the high level of perforin RNA. The membrane phenotypes of the final cell population, assessed by a panel of monoclonal antibodies (MoAbs) indicate a mixed population comprising two cell types in variable proportions (i) NKH-1+, T cell receptor (TCR) alpha/beta-, TCR gamma/delta-, CD3-, Leu 23+; (ii) NKH-(+), TCR alpha/beta-, TCR gamma/delta+, CD3+, Leu 23+. This culture system may provide a tool for cellular and molecular studies on the mechanisms of anti-tumor cytotoxicity, as well as the basis for new adoptive immunotherapy protocols in advanced cancers.
authors.affiliation: Department of Hematology and Oncology, Istituto Superiore di Sanità, Roma, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Montesoro
authors.forename: D
authors.initials: D
authors.lastname: Bulgarini
authors.forename: A
authors.initials: A
authors.lastname: Carè
authors.forename: R
authors.initials: R
authors.lastname: Masciulli
authors.forename: G
authors.initials: G
authors.lastname: Giannella
authors.forename: G
authors.initials: G
authors.lastname: Mariani
authors.forename: P
authors.initials: P
authors.lastname: Samoggia
authors.forename: G
authors.initials: G
authors.lastname: Salvo
authors.forename: D
authors.initials: D
authors.lastname: Habetswallner
authors.forename: U
authors.initials: U
authors.lastname: Testa
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007375:Interleukin-1; D007376:Interleukin-2; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008285:Major Histocompatibility Complex; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012313:RNA / Q000032:analysis; D011948:Receptors, Antigen, T-Cell / Q000032:analysis; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D007375:Interleukin-1; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D052899:Pore Forming Cytotoxic Proteins; D011948:Receptors, Antigen, T-Cell; D054353:Perforin; D012313:RNA