Mouse Macrophages Carrying Both Subunits of the Human Interferon-γ (IFN-γ) Receptor Respond to Human IFN-γ but Do Not Acquire Full Protection against Viral Cytopathic Effect

We have studied the prenuclear signal transduction pathway by which thyroid hormone potentiates the antiviral activity of human interferon-γ (IFN-γ) in HeLa cells, which are deficient in thyroid hormone receptor (TR). The action of thyroid hormone was compared with that of milrinone, which has structural homologies with thyroid hormone.l-Thyroxine (T4), 3,5,3′-l-triiodothyronine (T3), and milrinone enhanced the antiviral activity of IFN-γ up to 100-fold, a potentiation blocked by cycloheximide. The 5′-deiodinase inhibitor 6- n-propyl-2-thiouracil did not block the T4 effect. 3,3′,5,5′-Tetraiodothyroacetic acid prevented the effect of T4 but not of milrinone. The effects of T4 and milrinone were blocked by inhibitors of protein kinases C (PKC) and A (PKA) and restored by PKC and PKA agonists; only the effect of T4 was blocked by genistein, a tyrosine kinase inhibitor. In separate models, milrinone was shown not to interact with nuclear TR-β. T4 potentiation of the antiviral activity of IFN-γ requires PKC, PKA, and tyrosine kinase activities but not traditional TR.

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Tryptophan Depletion Modulates Tryptophanyl-tRNA Synthetase-Mediated High-Affinity Tryptophan Uptake into Human Cells

Genes ◽

10.3390/genes11121423 ◽

2020 ◽

Vol 11 (12) ◽

pp. 1423

Author(s):

Takumi Yokosawa ◽

Aomi Sato ◽

Keisuke Wakasugi

Keyword(s):

Interferon Γ ◽

Trna Synthetase ◽

Human Cells ◽

Human Interferon ◽

Tryptophan Depletion ◽

The Novel ◽

High Affinity ◽

Uptake Assay ◽

Ifn Γ ◽

Metabolizing Enzyme

The novel high-affinity tryptophan (Trp)-selective transport system is present at elevated levels in human interferon-γ (IFN-γ)-treated and indoleamine 2,3-dioxygenase 1 (IDO1)-expressing cells. High-affinity Trp uptake into cells results in extracellular Trp depletion and immune suppression. We have previously shown that both IDO1 and tryptophanyl-tRNA synthetase (TrpRS), whose expression levels are increased by IFN-γ, have a crucial function in high-affinity Trp uptake into human cells. Here, we aimed to elucidate the relationship between TrpRS and IDO1 in high-affinity Trp uptake. We demonstrated that overexpression of IDO1 in HeLa cells drastically enhances high-affinity Trp uptake upon addition of purified TrpRS protein to uptake assay buffer. We also clarified that high-affinity Trp uptake by Trp-starved cells is significantly enhanced by the addition of TrpRS protein to the assay buffer. Moreover, we showed that high-affinity Trp uptake is also markedly elevated by the addition of TrpRS protein to the assay buffer of cells overexpressing another Trp-metabolizing enzyme, tryptophan 2,3-dioxygenase (TDO2). Taken together, we conclude that Trp deficiency is crucial for high-affinity Trp uptake mediated by extracellular TrpRS.

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Human Interferon-γ(IFN-γ) Containing Cells Bear Various Surface Phenotypic Markers

Microbiology and Immunology ◽

10.1111/j.1348-0421.1986.tb03034.x ◽

1986 ◽

Vol 30 (10) ◽

pp. 1049-1059 ◽

Cited By ~ 5

Author(s):

Isamu Sugawara ◽

Hiroshi Kitagawa ◽

Hirofumi Inagaki ◽

Mark De Ley ◽

Akio Fukuda

Keyword(s):

Interferon Γ ◽

Human Interferon ◽

Phenotypic Markers ◽

Ifn Γ

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Assay of human interferon in Vero cells by several methods

Journal of Clinical Microbiology ◽

10.1128/jcm.9.4.471-475.1979 ◽

1979 ◽

Vol 9 (4) ◽

pp. 471-475

Author(s):

P C Ferreira ◽

M L Peixoto ◽

M A Silva ◽

R R Golgher

Keyword(s):

Vesicular Stomatitis Virus ◽

Sindbis Virus ◽

Reduction Method ◽

Cytopathic Effect ◽

Vero Cells ◽

Human Interferon ◽

Inhibition Assay ◽

Vesicular Stomatitis ◽

Viral Cytopathic Effect

Four methods for the assay of human interferon in Vero cells were compared based on the inhibition of viral cytopathic effect (CPE) in tubes, the inhibition of CPE in microplates, the reduction of plaques, and the inhibition of quantitative hemadsorption. For inhibition of CPE, Sindbis virus, vesicular stomatitis virus, poliovirus type 2, and vaccinia virus were used for challenge. In the plaque reduction method, Sindbis virus, vesicular stomatitis virus, and poliovirus were employed, and Newcastle disease virus was used in the quantitative hemadsorption assay. Sindbis virus was most susceptible to interferon in those tests measuring inhibition of CPE, but vesicular stomatitis virus was as sensitive in the plaque reduction method. Highest titers of interferon were recorded in microplates, especially with Sindbis virus as the challenge agent, followed by the quantitative inhibition assay. The CPE inhibition method was the simplest, and the quantitative hemadsorption assay was the most rapid to perform. Reproducibilities, as shown by the coefficient of variation, were 15, 39, and 59% for plaque reduction, CPE inhibition in tubes, and CPE inhibition in microplates, respectively.

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A Bivalent Immunoadhesin of the Human Interferon-γ Receptor Is an Effective Inhibitor of IFN-γ Activity

Journal of Interferon & Cytokine Research ◽

10.1089/jir.1995.15.1111 ◽

1995 ◽

Vol 15 (12) ◽

pp. 1111-1115 ◽

Cited By ~ 3

Author(s):

DIETER MOOSMAYER ◽

ELKE GERLACH ◽

ROLAND HAUFF ◽

PASCALE BECKER ◽

BODO BROCKS ◽

...

Keyword(s):

Interferon Γ ◽

Human Interferon ◽

Effective Inhibitor ◽

Ifn Γ

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High Throughput Screening for Human Interferon-y Production Inhibitor Using Homogenous Time-Resolved Fluorescence

CrossRef Listing of Deleted DOIs ◽

10.1177/108705710000500409 ◽

2000 ◽

Vol 5 (4) ◽

pp. 263-268 ◽

Cited By ~ 1

Author(s):

Koji Enomoto ◽

Yuko Aono ◽

Takashi Mitsugi ◽

Koji Takahashi ◽

Ryuji Suzuki ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Culture Supernatant ◽

Interferon Γ ◽

Interleukin 12 ◽

Human Interferon ◽

Time Resolved Fluorescence ◽

Time Resolved ◽

Ifn Γ

An immunoassay for interferon-γ (IFN-γ) using homogeneous time-resolved fluorescence (HTRF) has been developed. In this assay, IFN-γ can be detected by simply adding a mixture of three reagents-biotinylated polyclonal antibody, europium cryptate (fluorescence donor, EuK)-labeled monoclonal antibody, and crosslinked allophycocyanin (fluorescence acceptor, XL665) conjugated with streptavidin-and then measuring the time-resolved fluorescence. The detection limit of IFN-γ by the proposed method is about 625 pg/ml. We applied the method to the detection of IFN-γ secreted from NK3.3 cells and employed it in high throughput screening for IFN-γ production inhibitors. With this screening format, IFN-γ can be measured by directly adding the above reagents to microplate wells where NK3.3 cells are being cultured and stimulated with interleukin-12. This "in situ" immunoassay requires only pipetting reagents, with no need to transfer the culture supernatant to another microplate or wash the plate. Therefore, this screening format makes possible full automation of cell-based immunoassay, thus reducing cost and experimental time while increasing accuracy and throughput.

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Interaction of truncated human interferon γ variants with the interferon γ receptor: crucial importance of Arg-129

Biochemical Journal ◽

10.1042/bj3240591 ◽

1997 ◽

Vol 324 (2) ◽

pp. 591-595 ◽

Cited By ~ 11

Author(s):

Joost HAELEWYN ◽

Lieve MICHIELS ◽

Peter VERHAERT ◽

Marc F. HOYLAERTS ◽

Raphaël WITTERS ◽

...

Keyword(s):

Amino Acid ◽

Binding Capacity ◽

Interferon Γ ◽

Receptor Interaction ◽

Human Interferon ◽

Amino Acid Residues ◽

Surface Plasmon Resonance Analysis ◽

Resonance Analysis ◽

C Terminus ◽

Ifn Γ

Recombinant human interferon γ (IFN-γ), produced in Escherichia coli, was selectively truncated at its C-terminus with chymotrypsin, clostripain or plasmin. The C-terminal amino acid residues of the three truncated IFN-γ variants were identified as Phe136, Arg129 and Lys128, indicating the removal of 7, 14 and 15 amino acid residues from the full-length molecule. The absence of seven C-terminal residues did not influence the binding of IFN-γ to its receptor. In contrast, the truncation of 14 residues resulted in a decrease in the Ka value to 1/24, as determined by surface plasmon resonance analysis. The removal of one additional amino acid residue from the C-terminal region of IFN-γ led to a marked loss of receptor-binding capacity and biological activity. These observations demonstrate that Arg129 is an essential part of a functionally important C-terminal IFN-γ sequence that is involved in receptor interaction.

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Natural History and Evolution of Anti-Interferon-γ Autoantibody-Associated Immunodeficiency Syndrome in Thailand and the United States

Clinical Infectious Diseases ◽

10.1093/cid/ciz786 ◽

2019 ◽

Vol 71 (1) ◽

pp. 53-62 ◽

Cited By ~ 3

Author(s):

Gloria H Hong ◽

Ana M Ortega-Villa ◽

Sally Hunsberger ◽

Ploenchan Chetchotisakd ◽

Siriluck Anunnatsiri ◽

...

Keyword(s):

United States ◽

Natural History ◽

The United States ◽

Interferon Γ ◽

Mycobacterium Abscessus ◽

Annual Data ◽

Persistent Infections ◽

Ifn Γ ◽

Immunodeficiency Syndrome

Abstract Background The natural history of anti-interferon-γ (IFN-γ) autoantibody-associated immunodeficiency syndrome is not well understood. Methods Data of 74 patients with anti-IFN-γ autoantibodies at Srinagarind Hospital, Thailand, were collected annually (median follow-up duration, 7.5 years). Annual data for 19 patients and initial data for 4 patients with anti-IFN-γ autoantibodies at the US National Institutes of Health were collected (median follow-up duration, 4.5 years). Anti-IFN-γ autoantibody levels were measured in plasma samples. Results Ninety-one percent of US patients were of Southeast Asian descent; there was a stronger female predominance (91%) in US than Thai (64%) patients. Mycobacterium abscessus (34%) and Mycobacterium avium complex (83%) were the most common nontuberculous mycobacteria in Thailand and the United States, respectively. Skin infections were more common in Thailand (P = .001), whereas bone (P < .0001), lung (P = .002), and central nervous system (P = .03) infections were more common in the United States. Twenty-four percent of Thai patients died, most from infections. None of the 19 US patients with follow-up data died. Anti-IFN-γ autoantibody levels decreased over time in Thailand (P < .001) and the United States (P = .017), with either cyclophosphamide (P = .01) or rituximab therapy (P = .001). Conclusions Patients with anti-IFN-γ autoantibodies in Thailand and the United States had distinct demographic and clinical features. While titers generally decreased with time, anti-IFN-γ autoantibody disease had a chronic clinical course with persistent infections and death. Close long-term surveillance for new infections is recommended.

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The Role of Interleukine-10 and Interferon-γ as Potential Markers of the Evolution of African Swine Fever Virus Infection in Wild Boar

Pathogens ◽

10.3390/pathogens10060757 ◽

2021 ◽

Vol 10 (6) ◽

pp. 757

Author(s):

Sandra Barroso-Arévalo ◽

Jose A. Barasona ◽

Estefanía Cadenas-Fernández ◽

José M. Sánchez-Vizcaíno

Keyword(s):

Wild Boar ◽

Vaccine Candidate ◽

African Swine Fever Virus ◽

African Swine Fever ◽

Interferon Γ ◽

Fever Virus ◽

Control Group ◽

Challenge Virus ◽

Ifn Γ

African swine fever virus (ASFv) is one of the most challenging pathogens to affect both domestic and wild pigs. The disease has now spread to Europe and Asia, causing great damage to the pig industry. Although no commercial vaccine with which to control the disease is, as yet, available, some potential vaccine candidates have shown good results in terms of protection. However, little is known about the host immune mechanisms underlying that protection, especially in wild boar, which is the main reservoir of the disease in Europe. Here, we study the role played by two cytokines (IL-10 and IFN-γ) in wild boar orally inoculated with the attenuated vaccine candidate Lv17/WB/Rie1 and challenged with a virulent ASFv genotype II isolate. A group of naïve wild boar challenged with the latter isolate was also established as a control group. Our results showed that both cytokines play a key role in protecting the host against the challenge virus. While high levels of IL-10 in serum may trigger an immune system malfunctioning in challenged animals, the provision of stable levels of this cytokine over time may help to control the disease. This, together with high and timely induction of IFN-γ by the vaccine candidate, could help protect animals from fatal outcomes. Further studies should be conducted in order to support these preliminary results and confirm the role of these two cytokines as potential markers of the evolution of ASFV infection.

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THU0052 RELATIONSHIP BETWEEN INTERFERON-Γ-PRODUCING IMMUNOCOMPETENT CELLS AND DISEASE ACTIVITY IN ADULT-ONSET STILL’S DISEASE

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.1817 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 238.1-238

Author(s):

Y. Shimojima ◽

D. Kishida ◽

T. Ichikawa ◽

Y. Sekijima

Keyword(s):

T Cells ◽

Disease Activity ◽

Nk Cells ◽

Acute Phase ◽

Serum Ferritin ◽

Interferon Γ ◽

Adult Onset Still’S Disease ◽

Still’S Disease ◽

Ifn Γ ◽

Adult Onset Still's Disease

Background:In the acute phase of adult-onset Still’s disease (AOSD), elevated levels of proinflammatory cytokines including interferon-γ (IFN-γ) are shown. Moreover, IFN-γ impacts on activating macrophages which play a crucial role in the pathogenesis of AOSD. Natural killer (NK) cells and T helper cells are in charge of secreting IFN-γ in the innate and adaptive immune systems of disease, respectively. However, the features of their IFN-γ-producing variation depending on disease activity are still uncertain in AOSD.Objectives:We investigated characteristics of IFN-γ-producing CD4+T cells and NK cells in patients with AOSD.Methods:Twenty-four patients in the acute phase of AOSD (active AOSD), 8 of them after treatment (remission), and 12 healthy controls (HC) were recruited in this study. Peripheral blood mononuclear cells and serum samples were provided from them for the experimental analysis. Flow cytometry was used for analyzing CD4+T cells, CD4+regulatory T cells (Tregs), NK cells, and their intracellular IFN-γ expression levels as well as suppression assay of Tregs. The serum concentration of interleukin-18 (IL-18) was measured using commercially available ELISA kit. Relationship between the analyzed data and clinical findings related to disease activity were statistically evaluated.Results:IFN-γ expression in CD4+T cells was significantly higher in active AOSD than in HC (p < 0.05). Tregs also significantly indicated higher expression of IFN-γ in active AOSD than in HC (p < 0.0001); and moreover, Tregs were significantly impaired in their suppression ability (p < 0.05). In both CD4+T cells and Tregs, expression of IFN-γ was significantly correlated with serum ferritin levels in active AOSD (p < 0.05). IFN-γ expression in CD4+T cells was significantly higher in patients with splenomegaly than those without that (p < 0.05). The proportion of NK cells was significantly lower in active AOSD than in HC (p < 0.005), whereas IFN-γ expression in NK cells was significantly higher in active AOSD than in HC (p < 0.0005). The number of NK cells and IFN-γ-expressing NK cells had inverse relationship with serum ferritin levels in active AOSD (p < 0.05 and p < 0.005, respectively). Increased number of NK cells and their decreased expression of IFN-γ were significantly demonstrated in remission (p < 0.05). In the analyses of NK cell subsets, lower expression of IFN-γ in CD56brightNK cells and higher that in CD56dimNK cells were significantly indicated in active AOSD than HC (p < 0.05). In remission, IFN-γ expression was significantly decreased in CD56dimNK cells (p < 0.05) despite no significant recovery of that in CD56brightNK cells (p = 0.311). Meanwhile, increased expression of IFN-γ in CD56brightNK cells was demonstrated in only patients who were treated with biologics. Although serum levels of IL-18 were significantly higher in active AOSD than in remission and HC; however, they had no significant correlations with any analyzed data.Conclusion:CD4+T cells and NK cells promote IFN-γ expression in the acute phase of AOSD. Meanwhile, increased expression of IFN-γ in CD4+T cells and decreased number of NK cells were correlated with serum ferritin levels, suggesting that they are indicators of disease activity. Furthermore, high disease activity may impact on the alteration of IFN-γ-producing balance in two distinct population of NK cells, and the plasticity of Tregs leading to defect in suppression ability.Disclosure of Interests:None declared

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