Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) might be developed as a novel anti-tumor drug due to its selective cytotoxicity in tumor cells. The predicted Macaca mulatta TRAIL (mmTRAIL) is highly homologous to hTRAIL in nucleotide acid as well as amino acid sequence, suggesting that mmTRAIL might induce apoptosis of human cancer cells. However, the cytotoxicity of mmTRAIL in human cancer cells has not been investigated. In this paper, it is reported that the gene encoding mmTRAIL has been cloned by using reverse-transcriptase polymerase chain reaction (RT-PCR) from monkey peripheral blood mononuclear cells (PBMCs) in our laboratory. Subsequently, an expression plasmid was constructed by inserting mmTRAIL gene into pQE30 plasmid. After induction by addition of Isopropylβ-D-1-Thiogalactopyranoside (IPTG), mmTRAIL was expressed. MmTRAIL was recovered from supernatant of sonicated bacteria by Ni-NTA agarose affinity chromatography. SDS-PAGE and gel filtration chromatography demonstrated that mmTRAIL forms trimer in solution. In vitro assays indicated that mmTRAIL was cytotoxic to human COLO205 tumor cells but not to normal cells at low concentration of nanomole. In addition, antitumor effect of mmTRAIL was evaluated in mice bearing human COLO205 tumor xenografts. Intratumorally injected mmTRAIL significantly inhibited growth of tumor grafts. These results suggested that mmTRAIL was valuable as candidate drug for cancer-targeted therapy.
Objective To summarize and evaluate the significance of tumor necrosis factor (TNF) tolerance, and to understand the general characteristics and known molecular mechanisms of different forms of tolerance, including the roles of transcription factors, signaling systems and receptors. Method The relevant literatures at home and abroad in recent years were collected and readed, and the content related to TNF tolerance was summarized. ResultsTNF tolerance could be produced after TNF pretreatment, and be divided into absolute tolerance and induced tolerance. TNF tolerance was related to a variety of interrelated and interdependent intracellular signal transduction. There was cross tolerance between TNF and lipopolysaccharide. Conclusions TNF tolerance may represent a protective mechanism, participating in the termination of inflammation and preventing excessive or persistent inflammation. TNF tolerance may also trigger immune paralysis, leading to severe inflammatory diseases, such as sepsis. The understanding of TNF tolerance can promote the diagnosis of inflammation related diseases or the implementation of treatment methods, so as to achieve more accurate evaluation and treatment.
As a new treatment option after conventional corticosteroids and immunomodulatory drugs, biologics have been widely used in the clinical management of non-infectious uveitis in many countries due to their approved efficacy and safety. Anti-tumor necrosis factor-alpha monoclonal antibody is the most commonly used one. However, the guidance on its standardized application is lacking. The Ocular Immunology Group of Immunology and Rheumatology Academy in Cross-Straits Medicine Exchange Association compiled the Chinese expert consensus on treatment of non-infectious uveitis with anti-tumor necrosis factor-alpha monoclonal antibody. This evidence-based consensus is made according to the principle of consensus building and combines the clinical experience of the experts. Twelve recommendations are formatted on the application of Adalimumab and Infliximab. The interpretation of this consensus point will help improve the normative and effective application of anti-tumor necrosis factor-alpha monoclonal antibody in ophthalmologists, rheumatologists and immunologists.
Objective To investigate the effect of heme oxygenase 1 (HO-1) on the apoptosis of human degenerated nucleus pulposus (NP) cells induced by tumor necrosis factor α (TNF-α), and explore its possible molecular mechanism. Methods The intervertebral disc tissues were derived from patients with lumbar intervertebral disc herniation. Then, the NP cells were cultured in vitro and the third generation of NP cells were used for subsequent experiments. Cell counting kit 8 (CCK-8) method was used to observe the proliferative effect of TNF-α on the NP cells in vitro at the concentration of 10, 20, 50, 100, and 200 ng/mL. The most apropriate concentration was selected according to the result of CCK-8. The NP cells were cultured with basal medium (control group), TNF-α (TNF-α group), TNF-α and CoPP 10 μmol/L (CoPP group), and TNF-α and ZnPP 15 μmol/L (ZnPP group), respectively. After cultured, the cell poptosis was detected by Hoechst staining and flow cytometry; the expression of cleaved Caspase-3, epithelial membrane protein 1 (EMP-1), HO-1, and p-P65 proteins were detected by Western blot. In order to further explore the potential molecular mechanisms of HO-1 for cell apoptosis, the NP cells were cultured with TNF-α (TNF-α stimulated group), TNF-α and pyrrolidine dithiocarbamate (PDTC) 5 μmol/L (TNF-α+PDTC stimulated group), respectively. Then the cell apoptosis rate was measured by flow cytometry at 24 hours after cultured. Results The optimal concentration of TNF-α was 100 ng/mL. Hoechst staining showed that a few apoptotic cells could be observed in control group and CoPP group; the apoptosis-like nucleis were observed in TNF-α group and ZnPP group, which was the most significant in ZnPP group. Flow cytometry showed that the cell apoptosis rates of TNF-α group, CoPP group, and ZnPP group were significantly increased when compared with the control group (P<0.05). Compared with TNF-α group, the cell apoptosis rate in CoPP group decreased (P<0.05), while in ZnPP group it increased (P<0.05). Western blot showed that the expression of HO-1 protein in TNF-α group was decreased, and the expressions of cleaved Caspase-3, EMP-1, and p-P65 proteins were increased when compared with the control group (P<0.05). Compared with TNF-α group, the expression of HO-1 protein in CoPP group increased, and the expressions of cleaved Caspase-3, EMP-1, and p-P65 proteins were reduced (P<0.05); the expression of HO-1 protein in ZnPP group decreased (P<0.05), the expressions of cleaved Caspase-3 and EMP-1 proteins increased (P<0.05), and the expression of p-P65 protein was not significantly changed (P>0.05). Compared with TNF-α stimulated group, the cell apoptosis rate in TNF-α+PDTC stimulated group was significantly reduced (t=3.076, P=0.031). Conclusion HO-1 can inhibit the apoptosis of degerated NP cells induced by TNF-α, and its mechanism effect is by inhibiting the nuclear factor кB signaling pathway.
Objective To explore effects of macrophage migration inhibitory factor (MIF) inhibitor ISO-1 on intestinal injury in acute necrotic pancreatitis in pregnancy (ANPIP) rat. Methods Twenty-four pregnant SD rats were randomly averagely divided into three groups: a sham operation (SO) group, an ANP group, and an ANP model plus ISO-1 treatment group (ISO-1 group). A rat model of ANP was induced by the retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. The rats were killed and the inferior vena cava blood and the tissues of pancreas and jejunum were harvested at 12 h after the operation. The serum amylase (AMY), lipase (LIP), diamine oxidase (DAO), interleukin (IL)-1β, and IL-6 levels were measured. The pancreatic and jejunal tissues were taken for the pathological examination scoring. The immunohistochemical method was used to detect the expression of the MIF, nuclear factor (NK)-κB, or tumor necrosis factor (TNF)-α protein. Results ① Compared with the SO group, the serum AMY, LIP, DAO, IL-1β, and IL-6 levels were increased in the ANP group (P<0.050), which in the ISO-1 group were decreased as compared with the ANP group, the DAO, IL-1β, and IL-6 levels had significant differences (P<0.050), but the AMY and LIP levels had no significant differences (P>0.050). ② The pathological points of the pancreas and jejunum tissues were increased in the ANP group as compared with the SO group, which were significantly decreased in the ISO-1 group as compared with the ANP group (P<0.050). ③ The average integrated optical density divide by area of the NF-κB,TNF-α, and MIF were significantly increased in the ANP group as compared with the SO group, which were significantly decreased in the ISO-1 group as compared with the ANPgroup (P<0.050). Conclusions MIF inhibitor ISO-1 could protect intestinal injury in ANPIP rat. It is suggested that MIF is one of mechanisms in ANPIP with intestinal injury and might be correlated with activities of TNF-α and NF-κB.
Objective To investigate the effects of one-lung ventilation time on the concentration of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in the bronchoalveolar lavage fluid (BALF), serum inflammatory markers and early pulmonary infection after radical resection of esophageal cancer. Methods Ninety patients with thoracoscope and laparoscopic radical resection of esophageal carcinoma were chosen. According to the thoracoscope operation time, the patients were divided into 3 groups including a T1 (0.5–1.5 hours) group, a T2 (1.5–2.5 hours) group and a T3 (>2.5 hours) group. Immediately after the operation, the ventilated and collapsed BALF were taken. Enzyme-linked immunosorbent assay (ELISA) method was used to determine the concentration of IL-6 and tumour necrosis TNF-α. The concentrations of procalcitonin (PCT), C-reactive protein (CRP), and white blood cell (WBC) were measured on the first, third, fifth day after operation. The incidence of pulmonary infection was observed within 3 days after operation. Result The IL-6 values of the right collapsed lung in all groups were higher than those in the left ventilated lung. The TNF-α value of the right collapsed lung in the T2 group and T3 group was higher than that in the left ventilated lung (P<0.05). Compared with in the right collapsed lung, the TNF-α and IL-6 values gradually increased with the the duration of one-lung ventilation (P<0.05). Compared with the left ventilated lung groups, the IL-6 value increased gradually with the duration of one-lung ventilation time (P<0.05). The TNF-α value of the T3 group was higher than that of the T1 and T2 groups (P<0.05). The PCT value of the T3 group was higher than that of the T1 group and T2 group on the third, fifth day after operation (P<0.05). But there was no significant difference in CRP and WBC among the three groups at different time points. The incidence of pulmonary infection in the T3 group was significantly higher than that in the T1 group within 3 days after operation (P<0.05). Conclusion With the extension of one-lung ventilation time, the release of local and systemic inflammatory mediators is increased, and the probability of pulmonary infection is higher.
Objective To study the variety and the action of inflammatory cytokines and the relevant anti-inflammatory factors in acute pancreatitis (AP). Methods The authors observed the change of peripheral blood IL-6 and sTNFR in 41 patients with mild and severe AP in two groups on 1, 5, 14d after acute attack by ELISA. Results All cases recovered gradually in mild group (n＝22) after five days. Twelve patients improved gradually in severe group (n＝19) after 5-7 days. The level of sTNFR increased markedly in 2 groups at 1, 5, 14d(Ｐ＜0.001), and that of the severe group was markedly higher decreased gradually (Ｐ＜0.01). The level of IL-6 increased apparently only in severe group on 1d, 40.38 pg/ml∶12.4 pg/ml, (Ｐ＜0.001). The levels of IL-6 and sTNFR correlated respectively with severity of AP. Conclusion These results show that peripheral blood IL-6 and TNFα are useful index to supervise the severity and conversion and final results of AP.
Objective To explore the effectiveness of the transforming growth factor-β1(TGF-β1) and tumor necrosis factor-α(TNF-α) inducing human bronchial epithelial(HBE) cells to optimize epithelia-mesenchymal transformation(EMT) model. Methods Blank control, TGF-β1 10 ng/ml, TNF-α 10 ng/ml, TGF-β1 10 ng/ml+TNF-α 10 ng/ml induced human epithelial cells for 24 hours. Then the change of morphological alteration were observed by applying CCK8, cells migration assay and Western blot technique. Results When TGF-β1 plus TNF-α induced human epithelial cells for 24 hours, most of HBE cells traits changed including morphological alteration from cobblestone to fusiform, connection between cells vanishing, intercellular space broadening. In the experiments of checking cell migration capacity by the vitro scratch test, the group spacing was 420.06±10.38 μm in the blank control group, 499.86±34.00 μm in the TGF-β1 10 ng/ml group, 514.93±10.56 μm in the TNF-α 10 ng/ml group, 569.68±33.58 μm in the TGF-β1 10 ng/ml+TNF-α10 ng/ml group. TGF-β1 cooperated with TNF-α led to scratch spacing narrowing significantly. Western blot analysis showed that expression of E-cadherin and Vimentin varied significantly in the TGF-β1+TNF-α group. Conclusion Inducing human bronchial epithelial cell by TGF-β1 cooperated with TNF-α optimizes EMT model.
Anti-tumor necrosis factor-α monoclonal antibody agents have been widely applied in the management of autoimmune diseases. Among them, Adalimumab and Infliximab have been used for years in clinical practice in treating non-infectious uveitis and achieved satisfactory effects and safety. However, no guideline or expert consensus for their usage is available in China currently. It hopefully promotes standardized clinical application of anti-tumor necrosis factor -α monoclonal antibody in treating non-infectious uveitis, together with other senior experts in uveitis, the Ocular Immunology Group of Immunology and Rheumatology Academy in Cross-Straits Medicine Exchange Association form this evidence-based recommendations for clinicians’ reference.
The incidence of depression in patients with rheumatoid arthritis is higher. The concomitant depression will increase medical expense, reduce drug efficacy, lower its compliance, increase the incidence of complication, and affect the cure of rheumatoid arthritis. The influence of depression to rheumatoid arthritis is usually ignored in clinical work. In recent years, the pertinence between depression and immune disease in pathogenesis is found in research: depression will increase the risk of immune diseases in activate inflammation as well as extend and promote the release of inflammatory factors. This article reviews research progress of correlation between depression and rheumatoid arthritis.