There is considerable evidence that IL-17 contributes to the inflammation associated with rheumatoid arthritis (RA). IL-17 is spontaneously produced by RA synovial membrane cultures and high levels of IL-17 were detected in the synovial fluid of patients with RA [10,11]. In collagen-induced arthritis (CIA), an animal model reminiscent in several aspects to RA, treat- ment with neutralizing anti-IL-17 antibody after the onset of arthritis reduces joint inflammation, cartilage destruction and bone erosion . Authors proposed that the mechanisms responsible for slowing the disease are suppression of pro- inflammatory cytokines, such as IL-1β, TNF-α and IL-6, and elimination of the additive/synergistic effects between IL-17 and these pro-inflammatory cytokines. In addition, mice genet- ically deficient in IL-17 or IL-17R were found to be less sus- ceptible for induction of CIA . In contrast, local IL-17 overexpression accelerates the onset of CIA and aggravates synovial inflammation . Evidence that IFN-γ regulates sus- ceptibility to CIA through suppression of IL-17 comes from the observation that mice of the prototypical CIA-susceptible strain DBA/1 demonstrate a high IL-17 and low IFN-γ cytokine profile as compared with CIA-resistant C57BL/6 mice . In addition, knocking out the IFN-γ gene renders the C57BL/6 mice susceptible to disease and switched their CD4 + T cell
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Representative tumors harvested from each group were processed for histological analyses. Pathologic ana- lysis (via H&E staining) found that there were large areas of necrosis in the tumor tissue treated with Ad-IFNγ, while few necrotic areas were observed in the Ad-LacZ- treated and PBS-treated tumors (data not shown). Cell proliferation was estimated by the immunohistochemical assessment of the nuclear antigen Ki-67. The results showed that Ki-67 was significantly lower in tumor tissue treated with Ad-IFNγ than those in Ad-LacZ group and PBS group. For apoptosis analysis, TUNEL positive ratios were significantly higher in Ad-IFNγ groups than those in Ad-LacZ group and PBS group. The representative pictures for Ki-67 and TUNEL staining in PBS group, Ad-LacZ group, and 1×10 9 pfu Ad-IFNγ group were shown in Figure 6.
We systematically searched all studies that estimated the accuracy of the IGRA (using two commercial IGRAs: QFT-IT and T-SPOT) for ATB among HIV-seropositive adult individuals. We searched the EMBASE, Cochrane and MEDLINE databases to find research articles pub- lished from January 1, 2000 to October 30, 2015. Searches were implemented using combinations of the following terms: “ tuberculosis ” , “ active tuberculosis ” , “ interferon- gamma release assay ” , “ QFT-IT ” , “ T-SPOT ” , “ CFP-10 ” , “ ESAT-6 ” , “ IGRA ” , “ acquired immunodeficiency syn- drome ” , “ human immunodeficiency virus ” and “ HIV ” . We searched for additional references from review articles, guidelines and conferences when necessary.
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RESULTS. Children of parents who smoked between the prenatal period and year 11 were more likely to be in lower quartiles of interferon ␥ production than children of nonsmoking parents. In addition, maternal, paternal, and parental pack-years showed significant inverse dose-response relationships with interferon ␥ production in the child. These dose-response relationships with interferon ␥ remained significant for both paternal and parental pack-years among children of mothers who did not smoke during pregnancy, suggesting the existence of specific postnatal effects of environmental tobacco smoke exposure. In contrast, no significant effects of parental smoking were found on interleukin 4 production.
The biological mechanisms responsible for the onset and exacerbation of asthma symptoms in children may involve the epigenetic regulation of inflammatory genes after environmental exposures. Using buccal cells, we hypothesized that DNA methylation in promoter regions of two asthma genes, inducible nitric oxide synthase (iNOS) and interferon g (IFNg), can vary over several days. Repeat buccal samples were collected 4 to 7 days apart from 34 children participating in the Columbia Center for Children ’ s Environmental Health (CCCEH) birth cohort study. Several field duplicates (sequential collection of two samples in the field) and replicates (one sample pyrosequenced twice) also were collected to ensure consistency with collection and laboratory procedures. DNA methylation was assessed by pyrosequencing a PCR of bisulfite-treated DNA. We found that replicate and field duplicate samples were correlated strongly (r = 0.86 to 0.99, P < 0.05), while repeat samples demonstrated low within-subject correlations (r = 0.19 to 0.56, P = 0.06 to 0.30). Our data reveal DNA methylation as a dynamic epigenetic mechanism that can be accessed safely and reproducibly in an inner city pediatric cohort using non- invasive buccal swabs and pyrosequencing technology.
Whereas recent evidence implicates interleukin 22 (IL- 22) in the pathogenesis of skin disease in Ps [6,7], PsA has been postulated to more likely involve IL-17 [8,9]. Both cy- tokines can be made by the T helper 17 (Th17) cell subset, but recent reports have described T cells that make IL-22 alone [10,11]. These T cells, subsequently termed Th22 cells, produce IL-22 without IL-17 or interferon γ (IFNγ) and are characterized by the expression of certain chemo- kine receptors, including chemokine receptor 4 (CCR4) and CCR6, which can influence homing to skin and joints . Both Th17 and Th22 cells are influenced by the cyto- kine IL-23, which is required for their expansion and
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A plant viral vector engineered from an in vivo infectious clone of zucchini yellow mosaic virus (ZYMV) was used to express the human interferon-gamma (INF-γ) in planta. The INF-γ gene was in frame inserted between the P1 and HC-Pro ORFs of the ZYMV vector. The infectious activity of the vector was approved by rubbing the plasmid on Chenopodium quinoa and observing local lesions. Individual lesions were mechanically transferred to the systemic host plant zucchini squash at the stage of cotyledonary leaf. The stability of INF-γ expression was assessed by successive passages of recombinant viruses from infected plant and throughout the period of 35 days after inoculating in a single plant. Then, the leaf tissues of inoculated plant were analyzed for the presence of transgene by RT-PCR and western blot analysis. The recombimamt protein was purified using affinity chromatography method. The results showed approximately 1–1.2 mg INF- γ per 100 g tissues were purified from leaves two weeks post inoculation. Also, the vector was remarkably stable in squash after six serial passages and 35 days. The procedure provides a convenient and fast method for production of large quantities of pure INF-γ in planta. The system also has a potential for production of other proteins of interest in cucurbits to use as immunogen to produce antiserum or use for other purposes.
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vation of T helper cells producing IFN-γ, the primary switch factor for IgG2a. Following activation by mAb’s to CD3 and CD28, many IFN-γ–producing T cells are present in the spleen of vaccinated mice. These cells contribute to tumor inhibition in several ways other than induction of the Ig isotype switch. Intratumoral release of IFN-γ trig- gers a strong delayed-type hypersensitivity–like reaction with the recruitment of many reactive leukocytes and dendritic cells that is very efficient in tumor debulking and induction of a tumor-specific immune memory (44, 45). Moreover, IFN-γ released by activated T cells changes the genetic program of tumor cells overexpressing the HER-2/neu oncogene and inhibits their proliferation as well as their production of VEGF, while activating their production of the antian- giogenic monokines IP-10 and MIG (9). PCA shows that IFN-γ and IFN-γ–inducible genes are not upmodulated in the wk22pb glands (see additional information, ref. 23), even if upmodulation of the LAT gene (34) indicates the presence of a discrete amounts of T cells. How- ever, at 22 weeks of age (i.e., about 2 months after the boost), the reac- tive cell infiltration is reduced and transcription of IFN-γ and IFN- γ–inducible genes may have been already switched off and returned to basal levels.
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However, this paper has some limitations. For example, the sample size was not large, and only IL-17 and IFN-γ levels, γδΤ cells, and total T cell counts were measured, but relevant ani- mal experiments were not performed. Animal experiments were not conducted for the patho- genesis of lung cancer complicated with COPD, and changes in relevant factors during the pathogenetic process were not detected con- tinuously. Moreover, the pathogenesis mecha- nism was not deeply studied. However, this study can provide relevant ideas for the diagno- sis of lung cancer complicated with COPD. In conclusion, with the increased staging, the levels of IL-17 and IFN-γ were significantly increased, and there was no significant differ- ence between γδΤ cells and total T cell counts. The research results can provide a certain basis for early detection and diagnosis of lung cancer complicated with COPD.
In this work we explore differences in blood cells and cytokine profiles in children according to atopic status and asthma (atopic or non-atopic). The study involved measurement of Th1(IFN- γ ) and Th2 (IL-5 and IL-13) cytokines in Dermatophagoides pteronyssinus stimulated peripheral blood leukocytes, blood cell count, skin prick test and specific IgE against common aeroallergens. Atopic status was associated with eosinophilia and production of Th2 type cytokines. Atopic asthma was associated with eosinophilia and non-atopic asthma was associated with IFN- γ and elevated monocytes in blood. IFN- γ and monocytes might play a role in immunopathology of non-atopic asthma in Latin American children.
In this study, most IFNγ-p cats were negative to L. infantum antibody and DNA detection in blood (25/32, 78%). This finding means that the combination of sero- logical and molecular tests with L. infantum-specific IFN-γ evaluation offered a more accurate overall esti- mation of the exposure to L. infantum of cats under study. In fact, the prevalence of L. infantum was 5% for blood PCR and 22% for antibody detection but the combination of results obtained by serological and molecular testing did not increase the percentage of positivity because all PCR positive cats were also posi- tive for L. infantum antibodies (Fig. 1). However, when also considering positivity for L. infantum-specific IFN-γ production, the overall L. infantum prevalence raised to 36%. These data confirm that a consider- able percentage of the studied cats had contact with L. infantum and that detection of cell-mediated immune response by measuring specific IFN-γ production pro- vides a better estimation of exposure of cats to L. infan- tum in endemic areas as seen in dogs with different techniques [34, 36].
mitogen-activated protein kinase (MAPK) and NF-κB signaling pathways, which result in the expressions of many inflammatory genes including inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α and interleukin (IL)-6 and IL-12. IFN-γ, once known as macrophage activation factor, is produced by natural killer (NK) cells early in the immune response and later by type I T helper (Th1) cells. Binding of IFN-γ to its re- ceptor causes the activations of JAK1,2-STAT1, which enhance the expressions of IFN-γ-regulated genes in- cluding those required for antigen processing and pres- entation, antiviral state, and microbicidal functions in macrophages .
(clone UCHT1), PE-anti-CD69 (clone TP1.55.3; both from Beckman Coulter) and APC-anti-CD25 mAbs. Intracellular staining with APC-anti-interferon (IFN)-γ (clone B27; BD Biosciences) mAb was performed at days 5 and 6 of the co-culture, after a 4-h incubation of the cells with Golgi Stop (BD Biosciences) and IntraPrep Per- meabilization Reagent (see above). At least 5000–10,000 cells were obtained, measured using a 488-nm laser flow cytometer (FACSCalibur, BD Biosciences). Data were analyzed using CELLQuest ® software (BD Biosciences). The results are expressed as the percentage of labeled cells or as the ratio of mean fluorescence intensity (MFI) of specific labeling to background staining.
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IFNs: interferons; STAT1-CC: Ala-656 and Asn-658 sites with double-cysteine alternative in the SH2 domain of STAT1; IFN-γ: interferon-gamma; IFN-β: interferon-beta; JAK-STAT: the Janus activated kinase-signal transducer and activator of transcription; pSTAT1: STAT1 Tyr-701 phosphorylation; FBS: fetal bovine serum; CCK-8: cell counting kit-8; SDS-PAGE: SDS polyacrylamide gel electrophoresis; Ab: antibody; PVDF: polyvinylidene fluoride; PCNA: proliferat- ing cell nuclear antigen; ANOVA: analysis of variance; EGFP: enhanced green fluorescent protein; iNOS: inducible nitric oxide synthase; ISGs: interferon- stimulated genes.
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We presented a case of disseminated NTM infection in a previously healthy Asian male with anti- IFN-γ autoantibody. Detection of anti-IFN-γ autoantibodies may help to identify the risk of NTM infection, particu- larly in people of Asian origin. To date, there are no established differences in the treatments available for patients with or without anti-IFN-γ autoantibodies. Due to patient’s personal reasons, there was no serial data of titer of anti-IFN-γ autoantibodies in our case during treatment and clinical followup. However, dis- seminated NTM infection with anti-IFN-γ autoanti- bodies may indicate treatment failure and recurrent according to previous case reports. Rituximab may be considered as an optional treatment for refractory or recurrent NTM infection in patients with anti- IFN-γ autoantibody, although evidence to support this is currently lacking.
The patients included 57 males and 23 females aged 1-4 years old with an average of 2.24± 0.36 years. The course of the illness was 1-4 years with an average of 2.34±1.7 years. Me- anwhile, 80 infants and young children receiv- ing physical examinations in the same period were selected as the control group, including 52 males and 28 females aged 1-3 years old. The differences in gender, age, etc. of the two groups of children had no statistical signifi- cances (all P > 0.05). They were comparable. Detection of serum concentrations of IL-2 and IFN-γ
We studied serum cytokine profiles in this patient during the acute (days 4, 6), subacute (day 12) or convalescent phase (day 50) of JDM-MAS with IP. Multi-target streaming protein quantitative technology (BD-Pharmingen Cytometric Bead Array; BD Biosci- ences, Franklin Lakes, NJ, USA) was used to detect the serum cytokine levels of interleukin (IL)-2,4,6,10, tumor necrosis factor-α (TNF- α), and interferon-γ (IFN- γ), fol- lowing the manufacturer's instructions. The lower detec- tion limits for IL-2, IL-4, IL-6, IL-10, TNF-α, and IFN-γ
bipolar, and elongated cells detected in culture by rhodamine-phalloidin staining (Fig. 3c–e). We confirmed that cells are IBA1 positive (Fig. 4d–g). Moreover, we de- tected at the mRNA level the expression of CX3CR1 and CSF1-R (Fig. 4g–h), together with other specific markers, comprised in the recently characterized microglial signa- ture , i.e., P2RY12 and TMEM119 (Fig. 4h). The CCR2 transcript was undetectable (Fig. 4g), in line with previous observations [29, 38]. Consistently with the literature and the ATCC®‘s data sheet, the HMC3 cells were GFAP nega- tive (Fig. 5a, i.e., the dark gray histogram completely over- laps the background histogram). The glioblastoma U373 cell line, kindly provided by Prof. Grassi (Institute of Physiology, Catholic University Medical School, Rome, Italy), was used as a positive control for the anti-GFAP antibody (Fig. 3a). The activation marker MHCII (HLA-DR in the figure) was negative under basal condi- tion and upregulated in the 28% of the HMC3 cell popula- tion by IFNγ treatment (36 h, 10 ng/ml) (Fig. 5b). In addition, resting HMC3 cells were CD14 (Fig. 5c) and CD11b (Fig. 5d) negative, and these markers were not in- duced by IFNγ. With respect to CD14 expression, our re- sults are in agreement with the original characterization  and the evaluation performed in flow cytometry by Etemad and colleagues . Finally, the HMC3 (ATCC®CRL-3304) cells expressed CD68 under basal con- ditions (Fig. 5e), in agreement with previous observations [17, 29]. However, CD68 was expressed at low level indi- cated by a mean fluorescence intensity (MFI) of 3.3, and no further induction with IFNγ was detected (MFI = 3.6). As shown in Fig. 6a, the HMC3 (ATCC®CRL-3304) cells were found entirely and homogenously positive for human leukocyte antigens A, B, and C (MHCI antigens). The antibody used for this analysis is specific for the human
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The results of present study on the associations of the IFN-γ 874 T/A polymorphism with OLP are consistent with the functional effect of the IFN-γ 874 T/A poly- morphism as genotypes of the IFN-γ 874 T/A poly- morphism have been associated with low (AA), medium (AT), and high (TT) cytokine production . The IFN- γ 874 T/A polymorphism has been associated with auto- immune diseases in Caucasian, Latin American, and Middle Eastern, but not Asian, populations as shown in a recent meta-analysis . The reason why a particular polymorphism was found to be associated with one population and not another may be due to ethnic differ- ences in the frequency of IFN-γ 874 T/A polymorphism among these ethnic groups. The frequency distribution of genotypes of IFN-γ (874A/T) polymorphism in Table 3 Allele and genotype frequencies of TGF- β 1 (509C/T)
When we compared CXCL10 levels in SF to those in the peripheral circulation, we did observe a dramatic in- crease in CXCL10 in the majority of patients. In contrast, IFNγ levels were higher in serum, likely reflecting IFNγ produced from multiple sources, including psoriatic pla- ques . Although CXCR3 levels were not significantly different between SF and peripheral blood cells, there was also a strong positive correlation between CXCL10 and CXCR3 mRNA in SF cells, further supporting the notion that localized CXCR3-producing cells may be responsible for the concentration of CXCL10 in the SF. In two pa- tients measured, CXCL10 levels were reduced in SF com- pared with serum. The lower total SF volume, lower total nucleated cell count in SF, and lower polymorphonuclear cells in SF, as well as equivalent levels of total blood lymphocytes and monocytes, may have contributed to this difference. These observations could have led to reduced inflammatory cells overall, including CXCL10-expressing cells in the SF of these two patients.
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