concanamycin-a and Leukemia--Myeloid

The mechanism underlying the cytotoxicity mediated by a human CD4(+) cytotoxic T-lymphocyte (CTL) clone directed against a peptide derived from the acute myelogenous leukemia-associated fusion protein, DEK-CAN, was investigated. A DEK-CAN fusion peptide-specific CD4(+) Th0 CTL clone, designated HO-1, was established from the peripheral blood lymphocytes of a healthy individual. HO-1 exerted direct but not "innocent bystander" cytotoxicity within 2 hours. The cytotoxicity mediated by HO-1 was completely Ca2+-dependent. Because HO-1 lysed peptide-loaded Fas-deficient target cells derived from a patient with a homozygous Fas gene mutation, its cytotoxicity appeared to be mediated by a Fas-independent pathway. In addition, its cytotoxicity was only partially inhibited by treatment with concanamycin A and strontium ions, which are inhibitors of the perforin-based cytotoxic pathway. Although membrane-bound type of tumor necrosis factor-alpha (TNF-alpha) was expressed on HO-1, an anti-TNF-alpha antibody had no effect on HO-1-mediated cytotoxicity. HO-1 expressed mRNA for apoptosis-inducing mediators, including perforin, granzyme B, Fas ligand, TNF-alpha, and lymphotoxin; however, no DNA fragmentation was detected in target cells incubated with HO-1 by 5-[125I]Iodo-2'-deoxyuridine release assay and agarose gel electrophoresis of DNA. Although it has been suggested that the Fas/Fas ligand system is the main pathway by which CD4(+) CTL-mediated cytotoxicity is exerted in murine systems, HO-1 produced peptide-specific and HLA-restricted cytotoxicity via a Fas-independent and nonapoptotic pathway. The present study thus describes a novel mechanism of cytotoxicity mediated by CD4(+) CTL.

Topics: Acute Disease; Amino Acid Sequence; Anti-Bacterial Agents; Apoptosis; CD4-Positive T-Lymphocytes; Cytotoxicity, Immunologic; Fas Ligand Protein; fas Receptor; Gene Expression Regulation; Granzymes; Humans; Leukemia, Myeloid; Lymphokines; Lymphotoxin-alpha; Macrolides; Membrane Glycoproteins; Molecular Sequence Data; Oncogene Proteins; Oncogene Proteins, Fusion; Perforin; Pore Forming Cytotoxic Proteins; Recombinant Fusion Proteins; Serine Endopeptidases; Strontium; T-Lymphocytes, Cytotoxic; Tumor Necrosis Factor-alpha