BACKGROUND: The DR-70(TM) immunoassay is a newly developed cancer diagnostic test which quantifies the serum fibrin degradation products (FDP), produced during fibrinolysis, by antibody reaction. The purpose of this study was to evaluate the potential of DR-70(TM) immunoassay in screening malignant tumor. METHODS: Sample subjects were 4,169 adults, both male and female, who visited the health promotion center of a general hospital from March 2004 to April 2005 and underwent the DR-70(TM) immunoassay test and other tests for cancer diagnosis. The patient group was defined as 42 adults out of the sample subjects who were newly diagnosed with cancer during the same time period when the DR-70(TM) immunoassay test was performed. Final confirmation of a malignant tumor was made by pathological analysis. RESULTS: The mean DR-70(TM) level was 0.83+/-0.65 microgram/ml (range: 0.00 (0.0001)~7.42 microgram/ml) in the control group (n=4,127) as opposed to 2.70+/-2.33 microgram/ml (range: 0.12 ~ 9.30 microgram/ml) in the cancer group (n=42), and statistical significance was established (p<0.0001, Student t-test). When categorized by the type of malignant tumor, all cancer patients with the exception of the subgroups of colon and rectal cancer showed significantly higher mean DR-70(TM) levels compared with the control group (p<0.0001, Kruscal-Wallis test). The receiver operating characteristic (ROC) curve analysis revealed < or = 1.091 microgram/ml as the best cut-off value. Using this cut-off value, the DR-70(TM) immunoassay produced a sensitivity of 71.4%, a specificity of 70.1%, a positive predictability of 69.4%, and a negative predictability of 69.2% (1). CONCLUSION: A significant increase in the mean DR-70(TM) value was observed in the cancer group (thyroidal, gastric, breast, hepatic and ovarian) compared with the control group. In particular, the specificity and sensitivity of the DR-70(TM) immunoassay was relatively high in the subgroups of breast, gastric, and thyroidal cancer patients. There is need for further studies on a large number of malignant tumor patients to see how the DR-70(TM) level might be changed according to the differentiation grade and postoperative prognosis of the malignant tumor.
Adult ; Breast ; Colon ; Diagnosis ; Diagnostic Tests, Routine ; Female ; Fibrin Fibrinogen Degradation Products ; Fibrinolysis ; Health Promotion ; Hospitals, General ; Humans ; Immunoassay* ; Male ; Mass Screening ; Prognosis ; Rectal Neoplasms ; ROC Curve ; Sensitivity and Specificity ; Thyroid Gland

BACKGROUND: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. METHODS: Immunohistochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1beta and TNF-alpha in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. RESULTS: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proinflammatory cytokines such as IL-1beta and TNF-alpha. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. CONCLUSION: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.
Apoptosis ; Arthritis* ; Arthritis, Rheumatoid ; Blotting, Western ; Calcineurin* ; Cardiomyopathies ; Cyclosporine ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Humans ; Inflammation ; Interleukin-10 ; Interleukin-6 ; Matrix Metalloproteinases ; Osteoarthritis ; Stroke ; Synovial Membrane ; Transfection ; Tumor Necrosis Factor-alpha

BACKGROUND: Chronic inflammation in the brain has known to be associated with the development of a various neurological diseases including dementia. In general, the characteristic of neuro-inflammation is the activated microglia over the brain where the pathogenesis occurs. Pro-inflammatory repertoires, interleukin-1beta (IL-1beta) and nitric oxide (NO), are the main causes of neuro-degenerative disease, particularly in Alzheimer's disease (AD) which is caused by neuronal destruction. Those pro-inflammatory repertoires may lead the brain to chronic inflammatory status, and thus we hypothesized that chronic inflammation would be inhibited when pro-inflammatory repertoires are to be well controlled by inactivating the signal transduction associated with inflammation. METHODS: In the present study, we examined whether biphenyl dimethyl dicarboxylate (DDB), an active compound from Schizandra chinensis Baillon, inhibits the NO production by a direct method using Griess reagent and by RT-PCR in the gene expression of inducible nitric oxide synthase ((i)NOS) and IL-1beta. Western blots were also used for the analysis of NF-kappaB and IkappaB. RESULTS: In the study, we found that DDB effectively inhibited IL-1beta as well as NO production in BV-2 microglial cell, and the translocation of NF-kappaB was comparably inhibited in the presence of DDB comparing those to the positive control, lipopolysaccharide. CONCLUSION: The data suggested that the DDB from Schizandra chinensis Baillon may play an effective role in inhibiting the pro-inflammatory repertoires which may cause neurodegeneration and the results imply that the compound suppresses a cue signal of the microglial activation which can induce the brain pathogenesis such as Alzheimer's disease.
Alzheimer Disease ; Blotting, Western ; Brain ; Cues ; Dementia ; Gene Expression ; Inflammation ; Interleukin-1beta ; Microglia ; Neurons ; NF-kappa B* ; Nitric Oxide ; Nitric Oxide Synthase Type II ; Schisandra* ; Signal Transduction*

BACKGROUND: Rheumatoid arthritis (RA) is a chronic and systemic inflammatory disease that is characterized by invasive synovial hyperplasia, leading to progressive joint destruction. Recent studies have described that RA is caused by virus, bacteria or outside material. Approximately 2 to 20% of RA cases are reported to be associated with infected hepatitis C virus (HCV). However, the mechanisms underlying virus-induced RA are still unknown. Moreover, few molecular studies have addressed the inflammatory aspects of HCV-associated autoimmune RA. In this study, we aimed to determine whether or not another HCV core protein transactivates the IL-8 gene expression, prototypic chemokine, in synovial cell. METHODS: To establish the HCV core expressing stable synovial cell line, pCI-neo-core, a plasmid encoding HCV core protein, were transfected to HIG-82 cell line that is an established cell line from rabbit periaricular soft tissue. We examined the morphological changes and cell cycle distribution of HIG-82 cells with expression of HCV core protein by inverted microscopy and flow cytometry analysis, respectively. Also, we determined the mRNA levels of Interleukin (IL)-6 and IL-8 related to the inflammation by RT-PCR and then analyzed regulation of IL-8 expression by the NF-kB pathway. RESULTS: Our study showed no significant differences in morphology and cell cycle between HIG-82 control cell line and HIG-82 expressing HCV core protein. However, expression of HCV core protein induces the IL-8 mRNA expression in HIG-82 core cells via activated NF-kB pathway. CONCLUSION: These results suggest that HCV core protein can lead to enhanced IL-8 expression. Such a pro-inflammatory role may contribute to the etiologic pathogenesis in RA patients with HCV infection.
Arthritis, Rheumatoid ; Bacteria ; Cell Cycle ; Cell Line ; Flow Cytometry ; Gene Expression ; Hepacivirus* ; Hepatitis C* ; Hepatitis* ; Humans ; Hyperplasia ; Inflammation ; Interleukin-8* ; Interleukins ; Joints ; Microscopy ; NF-kappa B ; Plasmids ; RNA, Messenger

BACKGROUND: Granulocyte colony stimulating factor (G-CSF) is known as a cytokine central to the hematopoiesis of blood cells and to modulate their cellular functions. Besides granulocytes and their precursors, monocytes/macrophages and endothelial cells are direct target cells of G-CSF action. G-CSF influences immune cells in an anti-inflammatory way. METHODS: To evaluate whether G-CSF has a potential for preventing or ameliorating diseases characterized by mucosal inflammation, we used a mouse model with trinitrobenzene sulfonic acid (TNBS)-induced inflammatory colitis. To the mice model G-CSF was administrated daily by intraperitoneal injection. Macroscopic evaluation and immunohistochemical analysis of colonic tissues were performed. RESULTS: Recombinant human G-CSF significantly inhibited LPS-induced TNF-alpha mRNA expression in THP-1 cells. As for in vivo relevance, G-CSF dramatically reduced the weight loss of mice, colonic damage, and mucosal ulceration that characterize TNBS colitis. Moreover, G-CSF suppressed the expression of tumor necrosis factor-alpha, interleukin-1beta, and intercellular adhesion molecule-1 in TNBS colitis. CONCLUSION: Current results demonstrate that G-CSF may be an effective agent for the treatment of diseases characterized by mucosal inflammation.
Animals ; Blood Cells ; Colitis* ; Colon ; Colony-Stimulating Factors* ; Endothelial Cells ; Granulocyte Colony-Stimulating Factor ; Granulocytes* ; Hematopoiesis ; Humans ; Inflammation ; Inflammatory Bowel Diseases ; Injections, Intraperitoneal ; Intercellular Adhesion Molecule-1 ; Interleukin-1beta ; Mice* ; RNA, Messenger ; Tumor Necrosis Factor-alpha ; Ulcer ; Weight Loss

Type II collagen (CII), major component of hyaline cartilage, has been considered as an auto-antigen in rheumatoid arthritis (RA). However, the clinical and biological significances with regard to the CII autoimmunity need to be clarified in human RA. The presence of antibodies to CII has been identified in sera, synovial fluid, and cartilage of patients with RA. In our study, the increased titer of IgG anti-CII in sera was well correlated with C-reactive protein, suggesting that this antibody may reflect the inflammatory status of RA. The titer of anti-CII antibodies (anti-CII Abs) tended to be higher in early stages of diseases. In our extending study, among 997 patients with RA, 269 (27.0%) were positive for circulatory IgG antibody to CII, those levels were fluctuated over time. It is hard to assess the significant amount of T cell responses to CII and CII (255~274) in RA. By using a sensitive method of antigen specific mixed lymphocyte culture, we can detect the presence of CII-reactive T cells in peripheral blood mononuclear cells of RA patients. Sixty seven (46.9%) of 143 patients showed positive CII reactive T cell responses to CII or CII (255~274). The frequencies of CII reactive T cells were more prominent in inflamed synovial fluid (SF) than in peripheral blood. These T cells could be clonally expanded after consecutive stimulation of CII with feeding of autologous irradiated antigen presenting cells (APC). Moreover, the production of Th1-related cytokine, such as IFN-gamma, was strongly up-regulated by CII reactive T cells. These data suggest that T cells responding to CII, which are probably presenting the IFN-gamma producing cells, may play an important role in the perpetuation of inflammatory process in RA. To evaluate the effector function of CII reactive T cells, we investigated the effect of CII reactive T cells and fibroblasts-like synoviocytes (FLS) interaction on the production of pro-inflammatory cytokines. When the CII reactive T cells were co-cultured with FLS, the production of IL-15 and TNF-alpha from FLS were significantly increased (2 to 3 fold increase) and this increase was clearly presented in accord to the expansion of CII reactive T cells. In addition, the production of IFN-gamma and IL-17, T cell derived cytokines, were also increased by the co-incubation of CII reactive T cells with FLS. We also examined the impact of CII reactive T cells on chemokines production. When FLS were co-cultured with CII stimulated T cells, the production of IL-8, MCP-1, and MIP-1alpha were significantly enhanced. The increased production of these chemokines was strongly correlated with increase the frequency of CII reactive T cells. Conclusively, immune response to CII was frequently found in RA. Activated T cells in response to CII contributed to increase the production of proinflammatory cytokines and chemokines, which were critical for inflammatory responses in RA. The interaction of CII-reactive T cells with FLS further augmented this phenomenon. Taken together, our recent studies have suggested that autoimmunity to CII could play a crucial role not only in the initiation but amplification/perpetuation of inflammatory process in human RA.
Antibodies ; Antigen-Presenting Cells ; Arthritis, Rheumatoid* ; Autoimmunity ; C-Reactive Protein ; Cartilage ; Chemokine CCL3 ; Chemokines ; Collagen Type II* ; Cytokines ; Humans ; Hyaline Cartilage ; Immunoglobulin G ; Interleukin-15 ; Interleukin-17 ; Interleukin-8 ; Lymphocytes ; Synovial Fluid ; T-Lymphocytes ; Tumor Necrosis Factor-alpha

Oral administration of antigen has long been used in the induction of immune tolerance in various animal models of autoimmune diseases including rheumatoid arthritis (RA). Alleveation of arthritogenic symptoms has been reported from RA patients who received oral administration of type II collagen (CII) without side effects, however its rather inconsistent therapeutic efficacy and variation among patients calls for more detailed investigation on the mechanism of oral tolerance to be settled as regular treatment for RA. In an attempt to understand the immunogenic processes underpinning tolerance induction by orally administered CII, we analyzed changes in the expression of costimulatory molecules and STAT/SOCS signaling messengers in the mouse model of collagen induced arthritis (CIA). We found thatin the spleen of CIA mice, that has been undergone repeated oral feeding of CII prior to the induction of arthritis, showed increased promortion of CTLA4 expressing lymphocytes than in the spleen of PBS fed control. On the other hand, cells expressing CD28 or ICOS were decreased in the spleen of tolerized mice. Tolerance induction by oral CII administration also enhanced the expression of STAT6 in both RNA and protein level, while not affecting the expression of STAT3. The expression of SOCS3, which hasbeen known to transmit STAT-mediated signals from Th2 type cytokines, remained unchanged in the spleen of tolerized mice. Interestingly transcript of SOCS1, which has been associated with Th1 related pathways, was only visible in the spleen of tolerized but not of control mice, suggesting that as in the case of IL-6 signaling, it may exert a feed back inhibition toward the Th1 type stimulation.
Administration, Oral* ; Animals ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Collagen ; Collagen Type II* ; Cytokines ; Hand ; Humans ; Immune Tolerance ; Interleukin-6 ; Lymphocytes ; Mice* ; Models, Animal ; RNA ; Spleen

BACKGROUND: Psoriasis is an inflammatory skin disorder that is characterized by a marked proliferation of keratinocytes, vascular dilation and leukocyte infiltration. Cytokines play important roles in the pathogenesis of inflammatory disorders. An overexpression of proinflammatory cytokines was characterized in psoriasis plaque. Among these cytokines, IL-1beta is major pro-inflammatory cytokine synthesized during the infection and inflammatory process. The IL-1 receptor antagonist (IL-1Ra) competes for the same IL-1 receptor for IL-1alpha and -1beta, which prevents activation of the target cells. Three single nucleotide polymorphisms (SNPs) in the IL-1beta gene have been reported at position-31, -511 and +3954. Within the IL-1Ra gene (IL-1RN), there is a variable number of tandem repeats (VNTR) of an 86 bp length in intron 2. These polymorphisms related to cytokine production and associated with various diseases. METHODS: We investigated the polymorphisms of IL-1B (promoter -511 and +3954) and IL-1RN on 114 psoriasis patients and -311 healthy normal controls in Korean. We performed PCR-RFLP on single nucleotide polymorphisms (SNPs) of IL-1B (promoter -511 and +3954) and fragment analysis on IL-1RN 86 bp VNTR polymorphism. RESULTS: The frequency of IL-1B-511*1 allele (patients vs. controls; 50.0% vs. 42.3%, RR=1.4) was significantly increased and IL-1B -511*2 allele (patients vs. controls; 50.0% vs. 57.7%, RR=0.7) decreased in psoriasis patients compared to normal controls. We also analyzed the IL-1B -511 polymorphism according to patients' characters (age of onset, sex and family history). The IL-1B -511 alleles were significantly associated in patients with male and family history than health normal controls. There were no significant associations of IL-1B +3954 and IL-1RN polymorphisms with psoriasis patients. CONCLUSION: These results suggest that the polymorphism of IL-1B -511 could be genetic susceptibility to psoriasis in Koreans.
Alleles ; Cytokines ; Genetic Predisposition to Disease ; Humans ; Interleukin 1 Receptor Antagonist Protein ; Interleukin-1 ; Introns ; Keratinocytes ; Leukocytes ; Male ; Minisatellite Repeats ; Polymorphism, Single Nucleotide ; Psoriasis* ; Skin

BACKGROUND: The protective immunity against tuberculosis (TB) involves both CD4+ T cells and CD8+ T cells. In our previous study, we defined four Mycobacterium tuberculosis derived peptide epitopes specific for HLA-A*0201 restricted CD8+ T cells (ThyA30-38, RpoB127-135, 85B15-23, PstA175-83). In this study, we investigated the immune responses induced by these peptide specific CD8+ T cells in latently and chronically infected people with TB. METHODS: We characterized these peptide specific CD8+ T cell population present in PBMC of both TB patients and PPD healthy people using IFN-gammaelispot assay, intracellular staining and HLA-A2 dimer staining. RESULTS: The frequency of peptide specific CD8+ T cell was in the range of 1 to 25 in 1.7x10(5) PBMC based on ex vivo IFN-gamma elispot assay, demonstrating that these peptide specific CD8+ T cell responses are induced in both TB patients and PPD people. Short term cell lines (STCL) specific for these peptides proliferated in vitro and secreted IFN-gamma upon antigenic stimulation in PPD+ donors. Lastly, HLA-A*0201 dimer assays indicated that PstA175-83 specific CD8+ T cell population in PPD+ healthy donors is heterogeneous since approximately 25~33% of PstA175-83 specific CD8+ T cell population in PPD+ healthy donors produced IFN-gamma upon peptide stimulation. CONCLUSION: Our results suggest that MHC class I restricted CD8+ T cell mediated immune responses to M. tuberculosis infection are induced in both TB patients and PPD+ people; however, the CD8+ T cell population is functionally heterogeneous.
Cell Line ; Enzyme-Linked Immunospot Assay ; Epitopes ; HLA-A2 Antigen ; Humans ; Mycobacterium tuberculosis* ; Mycobacterium* ; Peptides ; T-Lymphocytes* ; Tissue Donors ; Tuberculosis*

BACKGROUND: Immunoglobulin (Ig) light chain repertoire has been implicated as a critical determinant in regulation of autoreactive B cells and production of pathogenic anti-DNA antibodies in systemic lupus erythematosus (SLE). We analyzed the impact of Ig lambda chain repertoire on development of autoimmunity in patients with SLE. METHODS: We obtained genomic DNA from individual peripheral CD19+ B cells of 3 untreated active SLE patients, and amplified Vlambda rearrangements from each single cell by polymerase chain reaction. RESULTS: A total number of 208 VlambdaJlambda rearrangements were analyzed. Analyzed sequences included 158 productive rearrangements and 50 nonproductive rearrangements. The differences in Vlambda gene usage in the productive and nonproductive repertoire of SLE patients were found compared to the non-autoimmune individuals. Vlambda gene, 9A was significantly overrepresented in nonproducative repertoire of SLE patients (P=0.016). In the productive repertoire, Vlambda genes, 3L and 1E were found more often in the SLE patients (P=0.001, P=0.043). When the productive and the nonproductive repertoires were compared, 9A was found significantly less in the productive repertoire in the SLE patients (P=0.000). There were no significant differences in the Jlambda gene usage between SLE patients and non-autoimmune individuals, but Jlambda2/3 gene was the most frequently used in SLE, whereas Jlambda7 gene was the most frequently used in the normal subjects. In the productive SLE Vlambda repertoire, 9.4% of the total sequences employed identical CDR3. It was particularly striking to find 7 identical versions of the 1G-Jlambda2/3 VlambdaJlambda rearrangements from one patient and 3 of the same sequence from another patient. Notably, identical Vlambda junctions in the SLE patients utilized significantly more homologous joining compared to Vlambda junctions of the normal adults (P=0.044). CONCLUSION: These data demonstrate regulation of lambda light chain expression in the SLE patients by selection of unique Vlambda genes. Also, biased selection and clonal expansion of particular Vlambda rearrangements are apparent in the SLE lambda repertoire.
Adult ; Antibodies, Antinuclear ; Autoimmunity ; B-Lymphocytes ; Bias (Epidemiology) ; DNA ; Humans ; Immunoglobulin Light Chains ; Immunoglobulins* ; Lupus Erythematosus, Systemic* ; Polymerase Chain Reaction ; Strikes, Employee