2.Screening of TNF-alpha antagonist peptides from a random peptide library displayed with Escherichia coli flagellar.
Chao LI ; Yan-Rong ZHOU ; Du-Sheng CHENG ; Pei-Tang HUANG
Chinese Journal of Biotechnology 2002;18(5):556-560
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
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genetics
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Peptide Library
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Peptides
;
isolation & purification
;
pharmacology
;
Tumor Necrosis Factor-alpha
;
antagonists & inhibitors
4.Juvenile psoriatic arthritis.
Shan LU ; Wei ZHOU ; Qian ZHANG ; Xin-Ying YU ; Dong-Ming LIU ; Xiang-Yuan LIU
Chinese Journal of Contemporary Pediatrics 2007;9(4):339-342
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
5.Patients treated with a tumor necrosis factor-alpha inhibitor are more likely to develop extrapulmonary tuberculosis.
The Korean Journal of Internal Medicine 2013;28(2):159-161
No abstract available.
Female
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Humans
;
*Immunocompromised Host
;
Immunosuppressive Agents/*adverse effects
;
Male
;
Tuberculosis/*immunology
;
Tumor Necrosis Factor-alpha/*antagonists & inhibitors
6.Prevention of TNF-induced necrotic cell death by rottlerin through a Nox1 NADPH oxidase.
Hee Sun BYUN ; Minho WON ; Kyeong Ah PARK ; Young Rae KIM ; Byung Lyul CHOI ; Hyunji LEE ; Jang Hee HONG ; Longzhen PIAO ; Jongsun PARK ; Jin Man KIM ; Gi Ryang KWEON ; Sung Hyun KANG ; Jin HAN ; Gang Min HUR
Experimental & Molecular Medicine 2008;40(2):186-195
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
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Animals
;
Benzopyrans/*pharmacology
;
Cell Death/*drug effects
;
Cell Line, Tumor
;
Mice
;
Protein Kinase Inhibitors/*pharmacology
;
Superoxides/metabolism
;
Tumor Necrosis Factor-alpha/*antagonists & inhibitors/pharmacology
7.Role of peroxisome proliferator-activated receptor beta in the inhibitory effect of epidermal growth factor on apoptosis of HaCaT.
Jie ZHOU ; Peng-fei LIANG ; Bi-mei JIANG ; Xiao-yuan HUANG
Chinese Journal of Burns 2009;25(4):294-297
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
8.Eukaryotic expression and bioactivity determination of the fusion protein sTNFRII-gAD consisting of soluble tumor necrosis factor receptor II and globular domain of adiponectin.
Suyun CHEN ; Qiushan HE ; Xiaoyan DONG ; Xiaobing WU ; Jimin GAO
Chinese Journal of Biotechnology 2010;26(2):207-215
In order to get soluble TNF receptor (sTNFR) II with good neutralizing activity against TNFalpha, we constructed the fusion gene sTNFRII-gAD, which encoded human sTNFR II and the globular domain of adiponectin (gAD), and then expressed it in mammalian cells and analyzed its anti-TNFalpha activity. First, sTNFRII cDNA was obtained by RT-PCR from the total RNA of human peripheral blood lymphocytes, and fused in frame with gAD gene. Then, the fusion gene sTNFRII-gAD was cloned into the expression vector pAAV2neo to result in the plasmid pAAV2neo-sTNFRII-gAD. By immunofluorescent staining with monoclonal antibody either against TNFRII or against adiponectin, we demonstrated that the pAAV2neo-s7NFRII-gAD-transiently-transfected BHK-21S cells were positive. To obtain G418-resistant BHK-21S/pAAV2neo-sTNFRII-gAD cells, we cultured the transfected BHK-21S cells above in 10% FBS containing DMEM media with 800 microg/mL G418 for 15 days, and changed the serum-containing culture media to a serum-free chemically defined media so as to change the cells culturing style from adhesion to suspension. 24 hours later, we harvested the supernatant of the culture for sTNFRII-gAD fusion protein characterization and anti-TNFalpha activity analysis. With monoclonal antibody either against TNFRII or against adiponectin, the Western blotting analysis showed that the sTNFRII-gAD fusion protein was expressed and existed as monomer, trimer and multimer forms in the supernatant. The bioactivity assay demonstrated that the sTNFRII-gAD fusion protein had the ability to neutralize TNFalpha so as to inhibit the cytotoxicity of TNFalpha on L929 cells. Put together, this study has laid the groundwork for large-scale preparation of sTNFRII-gAD fusion protein.
Adiponectin
;
biosynthesis
;
genetics
;
Animals
;
Cells, Cultured
;
Cricetinae
;
Humans
;
Protein Structure, Tertiary
;
genetics
;
Receptors, Tumor Necrosis Factor, Type II
;
biosynthesis
;
genetics
;
Recombinant Fusion Proteins
;
biosynthesis
;
genetics
;
pharmacology
;
Solubility
;
Tumor Necrosis Factor-alpha
;
antagonists & inhibitors
9.Effects of TNF-alpha receptor blocking peptide on adjuvant arthritis in rats.
Ya-ping HE ; Zhuo-ya LI ; Xiao-dan JIANG ; Wei FENG ; Yong XU ; Ping XIONG
Acta Pharmaceutica Sinica 2003;38(12):889-892
AIMTo study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.
METHODSThe model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.
RESULTSThe model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.
CONCLUSIONThe TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.
Animals ; Ankle Joint ; pathology ; Arthritis, Experimental ; immunology ; pathology ; Interleukin-1 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; metabolism ; Male ; Peptides ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics
10.A Case of Tuberculous Arthritis Following the Use of Etanercept.
Seung Won CHOI ; Jong Joon AHN ; Young Tae HWANG ; Sang Hoon KOH ; Sung Do CHO
The Korean Journal of Internal Medicine 2009;24(4):397-401
Etanercept is a tumor necrosis factor (TNF) inhibitor that has been used for the treatment of chronic inflammatory diseases including rheumatoid arthritis, ankylosing spondylitis and psoriatic arthritis. Because of its immunosuppressive activity, opportunistic infections have been noted in treated patients, most notably caused by Mycobacterium tuberculosis. Tuberculosis may present in an extrapulmonary or disseminated form. Since TNF-alpha inhibitors have been used in Korea, a few cases of TNF-alpha inhibitor associated tuberculosis have been described. However, tuberculous arthritis has not been previously reported. We describe a case of tuberculous arthritis in a 57-year-old woman with rheumatoid arthritis who was treated with etanercept.
Antirheumatic Agents/*adverse effects
;
Arthritis, Rheumatoid/drug therapy
;
Female
;
Humans
;
Immunoglobulin G/*adverse effects
;
Middle Aged
;
Receptors, Tumor Necrosis Factor
;
Tuberculosis, Osteoarticular/*chemically induced
;
Tumor Necrosis Factor-alpha/*antagonists & inhibitors