Humans ; Mucocutaneous Lymph Node Syndrome ; drug therapy ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; Tumor Necrosis Factors

Tumor necrosis factor(TNF-alpha) plays an improtant role in the process of anti-infection and anti-cancer. It can both protect and make damage to the host. In order to find new way of inhibiting its host-damaging activity, An E. coli flagella displayed random peptide library was constructed and TNF-alpha antagonist peptides were screened using the peptide library. After 5 rounds of panning and DNA sequencing, six peptide sequences were obtained. Two of them(TBP2, TBP3) have the same sequence frame of V--N-WG. After primary comparation of TNF-alpha binding ability, four peptides were synthesised and purified with RP-HPLC. The activity of inhibiting TNF-alpha was detected with L929 cell and MTT method. The data show that TBP2 and TBP3 can inhibit 90% of TNF-alpha activity when TNF-alpha gives about 30% cell toxicity on L929. The two sequences have not been reported.
Escherichia coli ; genetics ; Peptide Library ; Peptides ; isolation & purification ; pharmacology ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that regulate cell-matrix composition and are also involved in processing various bioactive molecules such as cell-surface receptors, chemokines, and cytokines. Our group recently reported that MMP-3, -8, and -9 are upregulated during microglial activation and play a role as proinflammatory mediators (Lee et al., 2010, 2014). In particular, we demonstrated that MMP-8 has tumor necrosis factor alpha (TNF-alpha)-converting enzyme (TACE) activity by cleaving the prodomain of TNF-alpha and that inhibition of MMP-8 inhibits TACE activity. The present study was undertaken to compare the effect of MMP-8 inhibitor (M8I) with those of inhibitors of other MMPs, such as MMP-3 (NNGH) or MMP-9 (M9I), in their regulation of TNF-alpha activity. We found that the MMP inhibitors suppressed TNF-alpha secretion from lipopolysaccharide (LPS)-stimulated BV2 microglial cells in an order of efficacy: M8I>NNGH>M9I. In addition, MMP inhibitors suppressed the activity of recombinant TACE protein in the same efficacy order as that of TNF-alpha inhibition (M8I>NNGH>M9I), proving a direct correlation between TACE activity and TNF-alpha secretion. A subsequent pro-TNF-alpha cleavage assay revealed that both MMP-3 and MMP-9 cleave a prodomain of TNF-alpha, suggesting that MMP-3 and MMP-9 also have TACE activity. However, the number and position of cleavage sites varied between MMP-3, -8, and -9. Collectively, the concurrent inhibition of MMP and TACE by NNGH, M8I, or M9I may contribute to their strong anti-inflammatory and neuroprotective effects.
Chemokines ; Cytokines ; Endopeptidases ; Inflammation ; Matrix Metalloproteinase Inhibitors* ; Matrix Metalloproteinases ; Microglia* ; Neuroprotective Agents ; Tumor Necrosis Factor-alpha*

Graft vs Host Disease ; drug therapy ; Humans ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; therapeutic use

A case of juvenile psoriatic arthritis in a 12 year-old boy was reported. The patient had a history of one and half a year of bilateral heel pain, followed by pain in the right knee and ankle and right hip joint. He developed psoriatic lesions affecting his nails and skin. He had increased erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) contents. Human leukocyte antigen (HLA) B27 was detected but serum rheumatoid factor was not in the patient. A skin biopsy revealed psoriasis and ultrasonography demonstrated synovitis in right knee and ankle. Juvenile psoriatic arthritis was diagnosed based on his physical, laboratory and skin biopsy findings. A treatment with nonsteroidal anti-inflammatory drugs and sulfasalazine produced no effect. Leflunomide in conjunction with anti-TNF biologic agents (Etanercept) was administered, followed by symptomatic improvement 2 weeks later.
Arthritis, Psoriatic ; diagnosis ; drug therapy ; etiology ; Child ; Diagnosis, Differential ; Etanercept ; Humans ; Immunoglobulin G ; therapeutic use ; Male ; Receptors, Tumor Necrosis Factor ; therapeutic use ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

Previous studies have demonstrated that rottlerin, a specific PKCdelta inhibitor, potentiates death receptor- mediated apoptosis through a cytochrome c-dependent or -independent pathway. However, its ability to regulate necrotic cell death, as well as the underlying mechanism, remains unknown. We found that in murine fibrosarcoma L929 cells, treatment with rottlerin protected the cells against TNF-induced necrosis, whereas it sensitized the cells to apoptosis induced by co-treatment with Hsp90 inhibitor geldanamycin and TNF, in a manner independent of its ability to inhibit PKC-delta. TNF treatment induced rapid accumulation of mitochondrial superoxide (O2") through the Nox1 NADPH oxidase when cells undergo necrosis. Moreover, pretreatment with rottlerin failed to induce the GTP-bound form of small GTPase Rac1 by TNF treatment, and subsequently suppressed mitochondrial O2(-) production and poly(ADP-ribose) polymerase activation, thus inhibiting necrotic cell death. Therefore, our study suggests that Nox1 NADPH oxidase is a new molecular target for anti-necrotic activity of rottlerin upon death-receptor ligation.
Acetophenones/*pharmacology ; Animals ; Benzopyrans/*pharmacology ; Cell Death/*drug effects ; Cell Line, Tumor ; Mice ; Protein Kinase Inhibitors/*pharmacology ; Superoxides/metabolism ; Tumor Necrosis Factor-alpha/*antagonists & inhibitors/pharmacology

No abstract available.
Female ; Humans ; *Immunocompromised Host ; Immunosuppressive Agents/*adverse effects ; Male ; Tuberculosis/*immunology ; Tumor Necrosis Factor-alpha/*antagonists & inhibitors

Histone acetylation plays a critical role in the regulation of transcription by altering the structure of chromatin, and it may influence the resistance of some tumor cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) by regulating the gene expression of components of the TRAIL signaling pathway. In this study, we investigated the effects and molecular mechanisms of trichostatin A (TSA), a histone deacetylase inhibitor, in sensitizing TRAIL-induced apoptosis in Caki human renal carcinoma cells. Our results indicate that nontoxic concentrations of TSA substantially enhance TRAIL-induced apoptosis compared with treatment with either agent alone. Cotreatment with TSA and TRAIL effectively induced cleavage of Bid and loss of mitochondrial membrane potential (MMP), which was associated with the activation of caspases (-3, -8, and -9) and degradation of poly (ADP-ribose) polymerase (PARP), contributing toward the sensitization to TRAIL. Combined treatment with TSA and TRAIL significantly reduced the levels of the cellular Fas-associated death domain (FADD)-like interleukin-1beta-converting enzyme (FLICE) inhibitory protein (c-FLIP), whereas those of death receptor (DR) 4, DR5, and FADD remained unchanged. The synergistic effect of TAS and TRAIL was perfectly attenuated in c-FLIP(L)-overexpressing Caki cells. Taken together, the present study demonstrates that down-regulation of c-FLIP contributes to TSA-facilitated TRAIL-induced apoptosis, amplifying the death receptor, as well as mitochondria-mediated apoptotic signaling pathways.
Acetylation ; Apoptosis* ; Caspases ; Chromatin ; Down-Regulation* ; Gene Expression ; Histone Deacetylase Inhibitors* ; Histones ; Humans ; Membrane Potential, Mitochondrial ; Tumor Necrosis Factor-alpha

Psoriasis is a chronic inflammatory disease. Medical therapy is the mainstay of the management of psoriasis, and the main target of psoriasis treatment is immunological dysregulation. Cyclosporine and methotrexate, the main conventional psoriasis treatments, usually lead to a Psoriasis Area and Severity Index (PASI) 75 response in 50% to 60% of patients, but show some organ toxicity. Biologics for psoriasis have recently become the main therapeutic agents for moderate to severe psoriasis unresponsive to conventional treatment. Tumor necrosis factor-α inhibitors were the first anti-psoriatic biologics to be developed, and also show good efficacy for psoriatic arthritis. Ustekinumab, the sole biologic designed for the inhibition of interleukin (IL)-12/23, has been most widely used for psoriasis in Korea. The main strength of ustekinumab is its relatively long treatment interval. IL-17 inhibitors have recently been introduced in Korea for psoriasis treatment. Secukinumab and ixekizumab are currently available IL-17 inhibitors that block the development of psoriasis lesions in the downstream events of psoriasis pathogenesis. They have excellent therapeutic efficacy, with a PASI 90 response in up to 60%–70% of patients. Selective IL-23 inhibitors have been more recently introduced in our country. They have an excellent PASI 90 response, and a longer injection interval than IL-17 inhibitors. New immunological modulators such as phosphodiesterase inhibitors, tyrosine kinase 2 inhibitors, and janus kinase inhibitors are planned to be introduced for psoriasis treatment. These are small molecules that can be administered orally, and some patients who are reluctant to receive injection therapy are expected to favor these therapeutic agents.
Arthritis, Psoriatic ; Biological Products ; Cyclosporine ; Humans ; Interleukin-17 ; Interleukin-23 ; Interleukins ; Korea ; Methotrexate ; Necrosis ; Phosphodiesterase Inhibitors ; Phosphotransferases ; Psoriasis ; Severity of Illness Index ; Tumor Necrosis Factor-alpha ; TYK2 Kinase ; Ustekinumab

OBJECTIVETo explore the role of EGF in regulating HaCaT apoptosis through peroxisome proliferator-activated receptor beta (PPARbeta).

METHODSCultured HaCaT cells were divided into different groups with different additives in culture medium as control (normal culture), TNF-alpha (with addition of 10 ng/mL TNF-alpha), EGF (with addition of 20 ng/mL EGF), EGF + TNF-alpha (cells were treated with 10 ng/mL TNF-alpha for 60 mins after the exposure to 20 ng/mL EGF for 4 hs) groups. Conjugation activity and transcription activity of PPARbeta of HaCaT cells in each group were detected by electrophoretic mobility shift assay (EMSA) and luciferase gene analysis (LGA). Protein expression of PPARbeta of HaCaT cells after transfected by missense oligonucleotide (scrODN) and antisense oligonucleotide (asODN) was determined by Western blot. Caspase-3 activity and apoptosis rate were detected by flow cytometry.

RESULTSConjugation and transcription activity of PPARbeta DNA were enhanced as shown in EMSA and LGA. Compared with that of cells in groups transfected by scrODN, protein expression of PPARbeta in cells of groups transfected by asODN was obviously inhibited as shown in Western blot. Caspase-3 activity of cells in TNF-alpha and EGF + TNF-alpha groups transfected by asODN was stronger than that of cells in TNF-alpha and EGF + TNF-alpha groups transfected by scrODN (P < 0.01). Apoptosis rate of cells in control, EGF, TNF-alpha, and EGF + TNF-alpha groups which were transfected by scrODN was (7.31 +/- 0.45)%, (7.43 +/- 0.21)%, (39.78 +/- 0.65)%, (28.34 +/- 0.54)% respectively, and that in those groups transfected by asODN was (8.22 +/- 0.51)%, (7.83 +/- 0.67)%, (46.78 +/- 0.48)%, (44.69 +/- 0.83)%. Apoptosis rate of cells in TNF-alpha and EGF + TNF-alpha groups transfected by asODN was respectively higher than that in TNF-alpha and EGF + TNF-alpha groups transfected by scrODN (P < 0.01).

CONCLUSIONSEGF inhibits HaCaT KC apoptosis caused by TNF-alpha in a PPARbeta-dependent manner.

Apoptosis ; drug effects ; Cell Culture Techniques ; Cell Line ; Epidermal Growth Factor ; pharmacology ; Humans ; PPAR-beta ; genetics ; metabolism ; Transcription, Genetic ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors