Pulmonary indoleamine 2,3-dioxygenase [indoleamine: oxygen 2,3-oxidoreductase(decyclizing)] has been found to be induced (30-to 100-fold) in the mouse after a single intraperitoneal administration of bacterial endotoxin [Yoshida, R. & Hayaishi, 0. (1978) Proc NatL AcadS Sci USA 75,[3998][3999][4000] [4084][4085][4086].In the present study, an in vitro system with mouse lung slices was developed in which bacterial endotoxin (5 ,ug/ml) produced an induction (approximately 10-fold) ofindoleamine 2,3-dioxygenase. The endotoxin was substituted by interferon from mouse L cells or mouse brain. The pulmonary enzyme activity increased almost linearly for 48 hr after addition of mouse interferon (104 units/ ml) to lung slices. Interferon from mouse L cells or mouse brain produced a 10-to 15-fold increase in the enzyme activity, whereas that from human leukocytes was all but ineffective. The effect also was observed using highly purified L-cell interferon, prepared by poly(U) affinity column chromatography. When interferon was treated either by heat, a'.chymotrypsin, or anti-interferon serum, such increase in the enzyme activity was diminished essentially to the same extent as seen in the antiviral activity. The increase in the enzyme activity was blocked when actinomycin D or cycloheximide was added to the slices before interferon treatment. These results suggest that the enzyme induction was produced by interferon and not by possible contaminants in the interferon preparations.Indoleamine 2,3-dioxygenase [indoleamine:oxygen 2,3-oxidoreductase (decyclizing)] (IDOase) is a hemoprotein (1) that catalyzes the incorporation of the superoxide anion as well as molecular oxygen (2, 3) into the pyrrole moiety of various indoleamine derivatives (4, 5). The pulmonary IDOase is dramatically induced (30-to 100-fold) in the mouse after a single intraperitoneal administration of bacterial endotoxin [lipopolysaccharide (LPS)] (6) or during virus infection (7). To determine the precise mechanisms of IDOase induction, we developed an in vitro system with lung slices from mice and examined the effects of various substances, including the superoxide anion, indoleamines, and interferons, on the enzyme activity. We report herein that the IDOase activity was increased approximately 10-to 15-fold within 48 hr after the addition ofmouse interferon (104 units/ml) to mouse lung slices. A species-specificity ofinterferons and the effects of inhibitors of protein synthesis are presented also.MATERILS AND METHODS Chemicals. L-[ring-2-'4C]Tryptophan (36 Ci/mol; 1 Ci = 3.7 x 1010 becquerels) was purchased from Commisariat a l'Energie Atomique, France, and was purified by column chromatography with Dowex 50W-X2 (H+ form) (0.5 x 1.0 cm), as described (8). LPS from Escherichia coli 055:B5, prepared by the Westphal method, was from Difco. LPS (2.5-500 .g/ml) was suspended in a nonpyrogenic, isotonic NaCl solution and was stored at 40C in a screw-capped vial. An aliquot (0.1 ml) of the suspension was added to Eagle's minimum essential mediu...

Granulysin has been identified as an effector molecule co-localized with perforin in the cytotoxic granules of cytotoxic T lymphocytes and natural killer (NK) cells, and has been reported to kill intracellular pathogens in infected cells in the presence of perforin and to induce a cytotoxic effect against tumor cells. The aim of the present study was to elucidate whether intracellular expression of granulysin and perforin by NK cells might be associated with progression of cancer. Flow cytometric analysis demonstrated high levels of perforin and granulysin expression by CD3(-) CD16(+) cells in healthy controls. In contrast, cancer patients exhibited significantly decreased levels of granulysin expression ( P<0.005), despite having equally high levels of perforin expression in comparison with healthy controls. The tumor-free patients expressed granulysin at levels similar to healthy controls, while the progressive tumor-bearing patients expressed remarkably lower levels of granulysin compared to healthy controls ( P<0.0001). Similarly, patients with an advanced performance status had significantly fewer granulysin-positive NK cells than healthy controls. Meanwhile, a considerable number of the tumor-bearing patients showed a decrease in the number of circulating NK cells, and a correlation between impaired granulysin expression and reduced circulating NK cells was observed. These findings suggest that the tumor-bearing patients with impaired granulysin expression were in an immunosuppressive state. In conclusion, impaired expression of granulysin by NK cells correlates with progression of cancer, and determination of granulysin expression might prove informative for assessing the immunological condition of cancer patients.

Oral administration of live Labre significantly increased IFN-alpha production in a dose-dependent manner. Labre intake tended to be most beneficial in subjects with initially low levels of IFN-alpha production. Heat-treated Labre did not elicit a response similar to that of the live bacteria.

To assess the clinical value of determination of the interferon (IFN)-producing capacity of patients, IFN production induced by Sendai virus (HVJ) in vitro was measured in cell cultures of whole blood from patients with various diseases. IFN production in patients with lung cancer, myelodysplastic syndromes, noninsulin-dependent diabetes mellitus, pulmonary tuberculosis, and asymptomatic HIV-1 infection was lower than that in healthy persons. Furthermore, periodic measurements of IFN production revealed decreasing IFN producing capacities in patients with lung cancer with progression of the tumor stage. However, increased IFN-producing capacities were observed in patients with tuberculosis after standard therapy. Further experiments showed that the main type of IFN induced in whole blood cultures was IFN-alpha, and decreased IFN production in patients did not result from a decreased number of leukocytes but rather from an impairment of cellular IFN production. The evaluation of IFN production in whole blood cell cultures may be a feasible method of assessing the impaired immune status.

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Inhibitors of fatty acid cyclooxygenase such as indomethacin (0.1 mM), phenylbutazone (0.3 mM), and aspirin (1 mM) were found to suppress almost completely the interferonmediated induction ofindoleamine 2,3-dioxygenase in mouse lung slices. However, phenacetin, an anti-inflammatory agent devoid ofcyclooxygenase inhibitory activity, and sodium salicylate, a weak inhibitor ofcyclooxygenase, were much less active under identical conditions. Glucocorticoids including dexamethasone, betamethasone, and cortisone, all of which are inhibitors of phospholipase A2, blocked induction of the dioxygenase by interferon in the nanomolar range, whereas other steroid hormones, such as aldosterone, testosterone, and 171-estradiol, were all but ineffective. These results suggest that the enzymes phospholipase A2 and fatty acid cyclooxygenase, both of which are essential for the biosynthesis of prostaglandins, play an important role in the induction of indoleamine 2,3-dioxygenase by interferon.

The effect of human natural interferons (IFN) alpha, beta, and gamma on the replication of human immunodeficiency virus (HIV) strain LAV in T cell lines TALL-1 and CCRF-CEM and peripheral blood lymphocytes (PBL) was studied. The growth of TALL-1 was moderately sensitive to these IFN, whereas that of CCRF-CEM was resistant to them. The progeny virus yield of LAV in TALL-1 at the time of its peak was reduced to 10% of the control level at IFN-alpha, beta, and gamma concentrations of 3, 11, and 23 IU/ml, respectively. These concentrations of IFN-alpha, beta, and gamma did not affect the cell growth. In CCRF-CEM, IFN-alpha, beta, and gamma at the concentration of 50, 60, and 76 IU/ml reduced the progeny virus to 10% of the control level. The virus growth in PBL was more sensitive to IFN than that in CCRF-CEM. The progeny virus yield was reduced to 10% of the control level by IFN-alpha and beta concentrations of 5 IU/ml and less than 5 IU/ml, respectively.