The Usefulness of In Vitro Interferon-gamma Assay for Differential Diagnosis between Intestinal Tuberculosis and Crohn's Disease / 대한소화기학회지

BACKGROUND/AIMS: It is difficult to clinically and endoscopically differentiate intestinal tuberculosis (ITB) and Crohn's disease (CD). The aim of this study was to evaluate the usefulness of in vitro interferon-gamma (INF-gamma) assay for differential diagnosis between ITB and CD. METHODS: Sixty patients for whom differential diagnosis between ITB and CD was difficult were enrolled between January 2007 and January 2009. The INF-gamma-producing T-cell response to early secreted antigenic target 6 and culture filtrate protein 10 were measured by T-SPOT.TB blood test in vitro. We evaluated the usefulness of T-SPOT.TB blood test by comparing its results with the final diagnosis. RESULTS: Twenty and forty patients were revealed to be positive and negative in T-SPOT.TB blood test, respectively. Of the 20 patients found to be positive, 12 patients (60%) were finally diagnosed as ITB, 6 patients as CD, and 2 patients as Behcet's enterocolitis. Of the 40 patients with negative results, 38 patients (95%) were diagnosed as CD; one as Behcet's enterocolitis; one as nonspecific colitis; none as ITB. The sensitivity and specificity of T-SPOT.TB blood test for ITB were 100% and 83.3%, respectively. Positive and negative predictive values of T-SPOT.TB blood test for ITB were 60.0% and 100%, respectively. CONCLUSIONS: When differential diagnosis between ITB and CD is difficult, T-SPOT.TB blood test may be a helpful and rapid diagnostic tool to exclude ITB. Prospective large-scaled studies are required for further evaluation of the usefulness of T-SPOT.TB blood test for differential diagnosis between ITB and CD.

The study on cytotoxicity of cytokines produced by the activated human NKT cells on neuroblastoma / 소아과

PURPOSE: alpha-Galactosylceramide (alpha-GalCer)-stimulated human Valpha24 natural killer T (NKT) cells exert antitumor activity against some leukemia in a CD1d dependent and TCR-mediated manner, but could not kill CD1d-negative neuroblastoma (NB) cells. There are few reports about the direct antitumor effect of highly secreted cytokines by these cells on activation. In this study, using a cell-free supernatant (SPN) collected from plate bound hCD1d/alphaGalCer tetramers-stimulated NKT cells, we examined whether they could be helpful in the immunotherapeutic treatment of NB. METHODS: Cells were cultured in IMDM. The cytokines produced by NKT cells were measured with Cytometric Bead Array (CBA) analysis. Cell viability was evaluated by calcein-AM fluorescence with digital image microscopy scanning (DIMSCAN). The percentage of specific apoptosis was calculated by flow cytometric detection of apoptosis using annexin V and 7-AAD. RESULTS: The activated NKT cells secreted high levels of IL-2, INF-gamma, TNF-alpha. The SPN was significantly cytotoxic against four out of eight tested NB cell lines, through mainly apoptosis as evidenced by annexin-V staining and inhibition with the pretreatment of pancaspase blocker. This apoptosis was significantly inhibited when anti-TNF-alpha and anti-IFN-gamma neutralizing mAbs were used separately and it was completely abolished when the two mAbs were combined. CONCLUSION: IFN-gamma and TNF-alpha produced by NKT cells could exert synergistically direct anti-tumor activity through apoptosis on some NB cell lines.

Markers of Inflammation in the Stenosis of Hemodialysis Vascular Access / 대한신장학회잡지

BACKGROUND: Histologically, vascular access stenosis is similar to the atherosclerotic lesion such as neointimal hyperplasia. However, the pathogenesis is not completely understood. The development of Inflammation and atherogenesis in hemodialysis population has been well recognized and is known as the malnutrition-inflammation-atherosclerosis (MIA) syndrome. We, therefore, examined the relationship between the vascular access stenosis and proinflammatory molecules in hemodialysis patients METHODS: Seventy-two regularly dialyzed patients were observed and the serum levels of the proinflammatory cytokines (IL-6, TNF-alpha and INF-gamma) were measured by ELISA. Patients were followed until the earliest date of death, modality change or loss to follow up. RESULTS: Vascular access stenosis was developed in thirty-five patients, and their mean duration of access survival was 22.7+/-37.6 months. In thirty-seven patients, vascular access remained patent during the study period (mean follow up; 70.9+/-46.6 months). In patients with autologous AV fistula, multivariate Cox regression analysis showed a significant contribution of INF-gamma to vascular access stenosis (Hazard ratio 1.024, p=0.044). In patients with AV graft, there was no significant relation between inflammatory cytokines and graft stenosis. There were positive correlations between the level of TNF-alpha and ferritin, and between transferrin and albumin. CRP and ferritin were negatively correlated with albumin and transferrin. CONCLUSIONS: Markers of inflammation were significantly associated with nutrition. And INF-gamma, one of the inflammatory cytokines, was related to the vascular access stenosis.

Markers of Inflammation in the Stenosis of Hemodialysis Vascular Access / 대한신장학회잡지

BACKGROUND: Histologically, vascular access stenosis is similar to the atherosclerotic lesion such as neointimal hyperplasia. However, the pathogenesis is not completely understood. The development of Inflammation and atherogenesis in hemodialysis population has been well recognized and is known as the malnutrition-inflammation-atherosclerosis (MIA) syndrome. We, therefore, examined the relationship between the vascular access stenosis and proinflammatory molecules in hemodialysis patients METHODS: Seventy-two regularly dialyzed patients were observed and the serum levels of the proinflammatory cytokines (IL-6, TNF-alpha and INF-gamma) were measured by ELISA. Patients were followed until the earliest date of death, modality change or loss to follow up. RESULTS: Vascular access stenosis was developed in thirty-five patients, and their mean duration of access survival was 22.7+/-37.6 months. In thirty-seven patients, vascular access remained patent during the study period (mean follow up; 70.9+/-46.6 months). In patients with autologous AV fistula, multivariate Cox regression analysis showed a significant contribution of INF-gamma to vascular access stenosis (Hazard ratio 1.024, p=0.044). In patients with AV graft, there was no significant relation between inflammatory cytokines and graft stenosis. There were positive correlations between the level of TNF-alpha and ferritin, and between transferrin and albumin. CRP and ferritin were negatively correlated with albumin and transferrin. CONCLUSIONS: Markers of inflammation were significantly associated with nutrition. And INF-gamma, one of the inflammatory cytokines, was related to the vascular access stenosis.

Circulating Levels of IFN-gamma, IL-10 and IL-16 in Patients with Alopecia Areata / 대한피부과학회지

BACKGROUND: Alopecia areata (AA) is an autoimmune condition of the hair follicle, resulting in bald patches. The details of the pathogenesis of AA still remain unclear. However, several recent studies have indicated that AA is an organ-specific autoimmune disease in which T cells (especially CD8+T cells), as well as certain cytokines (especially Th1 cytokines, IL-1, IFN-gamma, and TNF-alpha) may play an important role in its development. OBJECTIVE: The purpose of this study was to characterize the cytokine response in the peripheral blood of patients with AA, before and after treatment. METHODS: Twenty one active AA patients and 10 healthy people were evaluated in this study. The levels of 3 cytokines, including IFN-gamma, IL-10, and IL-16, in all subjects were measured at the first visit and 3 months after treatment. RESULTS: The levels of IFN-gamma, IL-10, and IL-16 in the AA group were significantly elevated (p75%), and was decreased in comparision to the level before treatment (p=0.003). There was no difference in the cytokine levels after PUVA, DPCP, or a combination therapy of PUVA and DPCP (p>0.05). CONCLUSION: The results suggest the involvement of IFN-gamma in the AA process. Also, IFN-gamma could be a potential marker for treatment. Even though different treatments have different mechanisms, IFN-gamma is considered to be a common pathway for alopecia areata treatment.

The Effects of Interferon -gamma on the Expression of Bullous Pemphigoid Antigen Genes in Cultured Keratinocytes / 대한피부과학회지

BACKGROUND: Interferon-gamma(IFN-gamma) has been shown to regulate epidermal keratinocyte growth and differentiation and can be isolated from subepidermal vesicles in bullous pemphigoid (BP) patients. In general, the IFN-gamma effects ate up-regulatory, and the corresponding transcriptional mechanisms have been elucidated in the case of several genes. One of the markers of the mitotic basal cell phenotype is the expression of BP antigens (BFAG), two hemidesmosomal proteins that were initially recognized as autoantigens in BP. OBJECTIVE: Since down-regulation of BPA gene expression is one of the earliest events during epidermal differentiation and BPAG is served as the target of autoantibodies, in this study, we examined the effect of IFN-gammaon the expression of the gene encoding the BPAG1 and 2. METHODS AND RESULTS: Northern analysis revealed a dose dependent suppression of BPAG expression by IFN-gamma in cultured human skin keratinocytes from 3 different donors, and incubation of the cells with IFN-gamma in the presence of cycloheximide demonstrated that this effect reqiured ongoing protein synthesis. Transient transfections of cultured keratinocytes with BPAG1 promoter-chloramphenicol acetyltransferase reporter gene plasmids indicated marked suppression of the promoter activity by IFN-gamma. CONCLUSION: This data, which inactivation of transcription of a basal keratinocyte-specific gene(BPAGl) by IFN-gamma. pravides new insight into the mechanisms of IFN-gamma mediated keratinocyte gene regulation and epidermal differentitation in inflammatory and blistering skin disease.

Cell Death of Corneal Fibroblasts Induced by Tumor Necrosis Factor-alphaand Interferon-gamma

PURPOSE: To evaluate the effect in the cell death of corneal fibroblasts when TNF-alphaand INF-gamma were given together. METHODS: Fibroblasts harvested from the human cornea were cultured in DMEM, then, nothing(control: Group 1), TNF-alphaonly(50 ng/ml : Group 2), INF-gammaonly(1.0 x 10(3)u/ml : Group 3), and a combination of both(Group 4) were added. We assessed the cell viability of the each group by the trypan blue exclusion assay at 4, 8, 12, 24, 48 hours after addition of cytokines. RESULTS: The cell viability at 48 hour after treatment was 94.27% in group 1, 90.68%(p=0.09) in group 2, 93.31%(p=0.45) in group3, and there was no statistical difference among the groups. Statistically signi-ficant decrease of the cell viability was achieved in group 4(82.86%, p=0.002). CONCLUSIONS: Cell death of human corneal fibroblasts had been increased after treatment with a combination of TNF-alphaand INF-gamma. These findings suggest that there could be some kind of interaction among the cytokines.