Effects of diethyldithiocarbamate, an inhibitor of interferon antiviral activity, upon human natural killer cells

• Published in: The Journal of immunology (1950) Vol. 132; no. 6; pp. 2868 - 2875
• Main Authors: Edwards, BS, Merritt, JA, Jelen, PA, Borden, EC
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Am Assoc Immnol 01-06-1984; American Association of Immunologists
• Subjects: 2',5'-Oligoadenylate Synthetase - metabolism; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Biological and medical sciences; Cytotoxicity, Immunologic - drug effects; Ditiocarb - pharmacology; Enzyme Activation - drug effects; Humans; Immunosuppressive Agents - pharmacology; Interferons - antagonists & inhibitors; Interferons - pharmacology; Killer Cells, Natural - drug effects; Killer Cells, Natural - enzymology; Killer Cells, Natural - immunology; Lymphocytes - drug effects; Lymphocytes - enzymology; Medical sciences; Pharmacology. Drug treatments; Sulfhydryl Reagents - pharmacology; Superoxide Dismutase - metabolism; Thiocarbamates - pharmacology; Natural killer cell; Investigation method; (2',5')A-Synthetase; Antiviral; Interferon; Mechanism of action; In vitro
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.132.6.2868

In support of a postulated role of the Cu++-dependent enzyme, superoxide dismutase (SOD), in antiviral effects of interferon (IFN), a close correspondence was previously shown to exist between inactivation of cellular SOD and concomitant blockade of IFN antiviral activity in fibroblasts by the Cu++-chelating agent, diethyldithiocarbamate (DDC). To further define the extent of "anti-IFN" activity, we initiated studies of DDC effects on IFN stimulation in the NK cell system. Unexpectedly, DDC directly inhibited cytotoxicity mediated by unstimulated NK cells. Pronounced inactivation occurred rapidly (less than 30 min), but was spontaneously reversible in the absence of DDC. Neither cell viability nor lymphocyte binding to target cells was detectably affected. Preincubation of DDC with Cu++ or Zn++ failed to neutralize its inhibitory effects nor could function be restored in DDC-pretreated NK cells by subsequent addition of Cu++, Zn++, Mg++, or Ca++. DDC treatment that inactivated NK cells did not detectably alter lymphocyte SOD activity. Thus, inhibition was probably not attributable to chelating properties of DDC. N-ethyl maleimide (NEM) and para-( hydroxymercuri ) benzoic acid ( PMBA ), enzyme inhibitors that preferentially react with sulfhydryl groups, both inactivated NK cells in a time- and dose-dependent manner similar to that of DDC. Preincubation with the sulfhydryl compound, cysteine, neutralized in parallel fashion the capacity of NEM, PMBA , and DDC to inhibit NK cell activity. Thus, a previously unreported reactivity of DDC with sulfhydryl groups appeared to be the basis of inhibition. NK cells incubated 1 hr with IFN and subsequently cultured 17 to 23 hr without IFN were activated to an extent comparable to cells continuously incubated 18 to 24 hr with IFN. Exposure to IFN for 1 hr was therefore sufficient to commit NK cells to acquisition of a fully activated state. Whether preactivated by a 1-hr or 18- to 24-hr IFN treatment, activated NK cells retained the DDC-sensitive phenotype characteristic of fresh unstimulated NK cells. Thus, prolonged IFN treatment did not render NK cells resistant to DDC or preferentially activate a DDC-sensitive NK cell subset. An 18- to 24-hr incubation of DDC-pretreated cells in the continual presence of IFN resulted in the boosting of NK cell activity. However, the 1-hr IFN pulse treatment protocol was consistently ineffective in boosting when IFN was added just after DDC-pretreatment. These results strongly suggested that DDC temporarily rendered NK cells unresponsive to what, under normal circumstances, approximated an optimally potentiating IFN stimulus.