No abstract available.
Immunoglobulin G ; Psoriasis ; Receptors, Tumor Necrosis Factor ; Etanercept

OBJECTIVEHuman soluble tumor necrosis factor receptor (sTNFR) can interfere with the biological functions of interleukin-1, which may be appropriate to the treatment of periodontitis. The eukaryote expression vector of the human sTNFR gene must be constructed prior to conducting transgene therapy of periodontitis.

METHODSBoth sTNFR gene and plasmid pcDNA 3.1 (+) DNA were digested with Kpn I and Xho I. After purification, the two fragments were ligated by TakaRa DNA Ligation Kit (Ver 2.0). This recombinant DNA was then transformed into E. Coli Competent Cells JM109 and positive clones were selected on the LB agarose plate containing ampicillin (80 microg/ul).

RESULTSSix single clones were indentified by double digestion with kpn I and xho I and two fragments with the size of 5.4 kb and 1.0 kb were produced as expected.

CONCLUSIONThe sTNFR gene was successfully inserted into the eukaryote expression vector plasmid pcDNA 3.1 (+) by recombination technology in vitro.

Tumor necrosis factor alpha (TNFalpha) is an intraovarian cytokine that may play a role in ovarian development and function. Identification of ovarian TNFalpha receptors provides support for establishing a role of TNFalpha in ovarian development and function. TNFalpha exerts its effects by binding to either TNF receptor 1 or 2 (TNFR1 or TNFR2). When TNFalpha binds with TNFR2, expression of survival genes is up-regulated, resulting in proliferation of granulosa cells. In the present study, the authors identified the changes in localization of TNFalpha and the expression of TNFR2 in granulosa cells during follicular atresia in rat ovaries. In healthy follicles, intense signals for TNFalpha and TNFR2 were found in the outer surface of the granulosa layer, where many proliferating cells and no apoptotic cells were observed. In atretic follicles, decreased expression of TNFalpha and TNFR2 was observed in the granulosa layer, where many apoptotic cells were seen. These findings suggested that TNFalpha acts as a survival factor in granulosa cells during follicular atresia in rat ovaries.
Animals ; Apoptosis ; Female ; Follicular Atresia ; Granulosa Cells ; Immunohistochemistry* ; Ovarian Follicle ; Ovary ; Rats* ; Receptors, Tumor Necrosis Factor ; Receptors, Tumor Necrosis Factor, Type II* ; Tumor Necrosis Factor-alpha*

Aspergillus fumigatus ; Cytokines ; Humans ; Interleukin-1beta ; Keratitis ; Receptors, IgE ; Tumor Necrosis Factor-alpha ; Tumor Necrosis Factors

PURPOSE: In prostate cancer, the anti-apoptotic mechanism of sulfated glycoprotein-2 (clusterin) against tumor necrosis factor-alpha (TNF-alpha) receptors and the action of type 2 TNF-alpha receptor (TNFR2) were investigated. MATERIALS AND METHODS: TNF-alpha, agonistic-TNF type 1 receptor (TNFR1) antibody, agonistic-TNF-R2 antibody and their combination were treated in PC3 cell line with or without anti-clusterin. Cytotoxicity was assessed by trypan blue dye exclusion assay. By using flowcytometric analysis, the exact amount of apoptosis and their changes were assessed. RESULTS: Apoptosis was significantly increased in both agonistic-TNFR1 antibody and TNF-alpha treated cases after blocking the activity of clusterin. The more the anti-clusterin antibody added, the more the apoptosis occurred. The increase of total apoptosis was greater in TNF-alpha treated cells than in agonistic-TNFR1 antibody treated ones. However, there was no increase of apoptosis in agonistic-TNFR2 antibody and TNF-alpha with agonistic-TNFR2 antibody treated cases, respectively. CONCLUSIONS: Clusterin prevents TNF-alpha induced apoptosis by affecting TNFR1. The difference in degree of apoptosis between agonistic-TNFR1 antibody treated cells and TNF-alpha treated ones suggests the possibility of the action of TNFR2. It may be associated with affinity of TNF-alpha to the tumor cell surface.
Apoptosis ; Cell Line ; Clusterin ; Diminazene ; Prostate ; Prostatic Neoplasms ; Receptors, Tumor Necrosis Factor ; Receptors, Tumor Necrosis Factor, Type I ; Receptors, Tumor Necrosis Factor, Type II ; Trypan Blue ; Tumor Necrosis Factor-alpha

PURPOSE: This study was performed to evaluate anti-apoptotic mechanism of Sulfated Glycoprotein-2 (SGP-2) in tumor necrosis factor-alpha (TNF-alpha) induced apoptosis of prostatic cancer cell. MATERIALS AND METHODS: To suppress the activity of natively secreted SGP-2 by prostatic cancer cells, varying amount of the blocking antibody to SGP-2 (anti-SGP-2 antibody, 1.25~2.0microgram/ml) was applied. Then TNF-alpha or agonistic antibody to type I TNF-alpha receptor (agonistic-TNFR1 antibody) were applied for the induction of apop tosis. The changes of cell number and cytotoxicity were assessed by trypan blue dye exclusion assay. The exact amount of early and late apoptosis were analyzed and compared by flowcytometric analysis. RESULTS: By blocking SGP-2 with anti-SGP-2 antibody (2.0microgram/ml), apoptosis was signi ficantly increased in both TNF-alpha (96.8 +/- 1.4%) and agonistic-TNFR1 antibody treated PC3 cells (95.2 +/- 0.9%) compared to control group (41.4 +/- 4.7%), especially after 36 hours incubation (p<0.05). The more suppressed the activity of SGP-2 by anti-SGP-2 antibody, significantly the more apoptosis was happened (p<0.05). After 24 hours incubation, the increase of apoptosis by the suppression of SGP-2 was significantly greater in TNF-alpha treated PC3 cells (45.6 +/- 27.6%) than in agonistic- TNFR1 antibody treated ones (28.6 +/- 4.7%). However after 36 hours the difference in degree of apop tosis between two groups disappeared. CONCLUSIONS: We found that SGP-2 blocked the apoptosis which was induced by agonistic-TNFR1 antibody as like TNF-alpha induced one. These results implies the possi bility of the direct action of SGP-2 on type I TNF-alpha receptor. However the difference between two groups in the degree of apoptosis after 24 hours incubation period sug gest the more efficient blocking effect of SGP-2 on agonistic-TNFR1 induced apoptosis than on TNF-alpha induced one. For clarification of these difference, further research about other influencing factors such as type II TNF-alpha receptor activation is essential.
Apoptosis* ; Cell Count ; Clusterin* ; Prostatic Neoplasms* ; Receptors, Tumor Necrosis Factor, Type I ; Trypan Blue ; Tumor Necrosis Factor-alpha

CD4⁺CD25⁺FoxP3⁺ regulatory T (Treg) cells play major roles in the maintenance of immune homeostasis. In this review, we comprehensively describe the relationship between tumor necrosis factor (TNF) and Treg cells, focusing on the effects of TNF on Treg cells and on TNF-producing Treg cells. Contradictory results have been reported for the effect of TNF on the suppressive activity of Treg cells. In patients with rheumatoid arthritis, TNF has been shown to reduce the suppressive activity of Treg cells. Meanwhile, however, TNF has also been reported to maintain the suppressive activity of Treg cells via a TNFR2-mediated mechanism. In addition, Treg cells have been found to acquire the ability to produce TNF under inflammatory conditions, such as acute viral hepatitis. These TNF-producing Treg cells exhibit T helper 17-like features and hold significance in various human diseases.
Arthritis, Rheumatoid ; Hepatitis ; Homeostasis ; Humans ; Inflammation ; Receptors, Tumor Necrosis Factor, Type II ; T-Lymphocytes, Regulatory ; Tumor Necrosis Factor-alpha

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Death receptors induce apoptosis by intracellular signaling transmission and relate to body growth, development, disease and death. This paper reviews the structural character of death receptors, and the mechanisms by which apoptosis is induced. The biology and pathobiology of these death receptors in liver are discussed as well.
Apoptosis ; physiology ; GPI-Linked Proteins ; Humans ; Liver ; pathology ; physiology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Receptors, Tumor Necrosis Factor ; physiology ; Receptors, Tumor Necrosis Factor, Member 10c ; Tumor Necrosis Factor Decoy Receptors ; fas Receptor ; physiology