TNF-α Monoclonal Antibody Research Articles

Development of a serum free medium for HUMIRA® biosimilar by design of experiment approaches

Abstract Background Serum have been traditionally used to support growth of animal cell cultures. However, the increasing growth of therapeutic biopharmaceuticals market, accelerated the high demand for the serum-free medium (SFM). Objective The main objective is to design a SFM for a stable rCHO cell line that produces a fully anti-human TNF-α monoclonal antibody (mAb) corresponding to HUMIRA® biosimilar. Materials and methods Design of Experiment (DoE) approaches were used to determine the key factors due to their effect on specific growth rate and mAb production. The production was carried out in T-flasks at different initial cell concentrations and then in Erlenmeyers with the developed SFM. mAb production was compared with commercial SFMs in terms of yield and productivity. Results Regarding to our findings, when the developed SFM-adapted cells were compared with the cells produced in commercial SFMs, the mAb productivity in developed SFM were higher (1.3–1.6 times) depending on higher mAb concentration and less (3–5 times) cell concentration. Additionally, the produced mAb in the developed SFM provided high conformational similarity with its originator HUMIRA®. Conclusion DoE approaches could be used to reduce cost and time in designing SFM for any commercially important cell line to produce high value biologics.

Diagnosis and Treatment Recommendation and Exploration for Critical and Refractory Adverse Effects Related to Immunocheckpoint Inhibitors

免疫检查点抑制剂（immune checkpoint inhibitors, ICIs）的应用改写了很多恶性肿瘤的治疗策略，成为肿瘤治疗的又一个里程碑。ICIs作用原理可以理解为“刹车理论”，在“松开刹车”后会带来一系列的全身性毒副反应，其中部分可能为危重和难治性，甚至具有潜在致死性。本文对免疫相关不良事件（immune-related adverse effects, irAEs）相关的最新国内外指南及共识中关于3度-4度irAEs的诊治建议进行了汇总，包括欧洲肿瘤内科学会年会（European Society for Medical Oncology, ESMO）、美国国立综合癌症网络（National Comprehensive Cancer Network, NCCN）/美国临床肿瘤学会（American Society of Clinical Oncology, ASCO）、肿瘤免疫治疗学会（Society for Immunotherapy of Cancer, SITC）和中国临床肿瘤学会（Chinese Society of Clinical Oncology, CSCO）指南，并复习了2019年5月20日前公开发表的关于irAEs的个案报告和相关的综述文献后，对以上指南和共识中尚未纳入的3级-4级irAEs的诊治建议进行补充，重点介绍了特异性免疫抑制药物在不同的irAEs中成功应用的情况，包括抗白介素6（interleukin 6, IL-6）阻断、抗CD20单抗、抗肿瘤坏死因子α（tumor necrosis factor-α, TNFα）单抗、抗整合素4单抗、Janus激酶抑制剂、血小板生成素受体激动剂和抗胸腺细胞球蛋白（antithymocyte globulin, ATG）。本文对于类固醇激素在irAEs中超大剂量使用和升级使用及反复使用提出质疑，并强调应同时关注激素继发的感染、肿瘤进展和无法满足ICIs再挑战的等问题。本文提出对于危重和难治性irAEs的“降阶梯治疗”原则，建议应尽早使用细胞因子靶向药物这些特异性免疫抑制药物。免疫治疗时代诸多的irAEs是传统化疗及小分子靶向治疗时代不曾有过的，不断地挑战肿瘤科大夫的知识储备和临床基本技能。因此，建立肿瘤多学科讨论体系对于肿瘤患者的治疗管理极为重要。

Adalimumab improves cognitive impairment, exerts neuroprotective effects and attenuates neuroinflammation in an Aβ1-40-injected mouse model of Alzheimer's disease

The pathogenesis of Alzheimer's disease (AD) is associated with an increased inflammatory response via activated microglia and astrocytes. In the present study, we investigated whether treatment with the anti–tumor necrosis factor alpha (TNF-α) monoclonal antibody adalimumab can improve cognitive function and reduce AD pathology in Aβ1-40-injected animal models of AD, as well as the mechanisms underlying the effects of treatment. Aβ1-40-injected mice treated with adalimumab exhibited significant improvements in memory relative to mice injected with Aβ1-40 alone, as well as decreases in beta secretase-1 (BACE1) protein expression and Aβ1-40 plaques. In addition, adalimumab treatment significantly attenuated neuronal damage and neuroinflammation in Aβ1-40-injected mice. Aβ1-40-induced decreases in brain-derived neurotrophic factor (BDNF) expression were also attenuated by treatment with adalimumab. Our experiments further verified that the effects of adalimumab are mediated by nuclear factor kappa B (NF-κB) p65 signalling. Serine 536 residues of NF-κB p65, which is phosphorylated by TNF-α, increased along with the degradation of inhibitor of κB (IκB) in the hippocampus of Aβ-injected mice, although these effects were again attenuated by adalimumab. Furthermore, Aβ1-40-induced increases in TNF-α and interleukin (IL)-6 expression were decreased by treatment with adalimumab. Our results indicate that adalimumab may be clinically useful in human patients with AD.

Incidence and Management of Infusion Reactions to Infliximab in an Alternate Care Setting

BackgroundInfliximab is a chimeric anti-tumor necrosis factor alpha (TNF-α) monoclonal antibody that ameliorates inflammation when it binds to and neutralizes TNF-α. It is often used in patients with Crohn’s disease and ulcerative colitis to reduce the severity of disease symptoms and induce disease remission. Infusions are generally administered in the hospital setting due to concerns over patient safety, and limited data exist regarding the incidence and management of infusion reactions (IRs) in an alternate care setting without direct physician oversight.AimsThe aim of this study was to evaluate the incidence of IRs following administration of infliximab and associated management approaches in an alternate care setting.MethodsA retrospective chart review of 796 patients with Crohn’s disease or ulcerative colitis that received a combined 5581 infusions with one home infusion provider between January 2014 and November 2016 was conducted. Timing, severity, management approach, and outcomes of IRs were abstracted and analyzed.ResultsA total of 109 infusion reactions (2.0% of all infusions) were recorded in 62 patients (7.8% of all patients). The majority of these reactions were acute and mild or moderate in severity and resolved with rate adjustments and/or medication. Emergency room visits were required in 0.1% of all infusions, and 0.3% of all infusions were not completed due to a reaction.ConclusionsIRs to infliximab were uncommon and mostly mild or moderate in severity. Resolution of the IR and continuation of therapy was achieved in most patients through a management approach that included prompt recognition and initial treatment via rate adjustments and medications according to physician’s orders.

Adolescent Female with Jaundice and Edema

An 18-year-old female presented to the emergency department at Texas Children's Hospital with intermittent symptoms of fever, nausea, poor food intake, fatigue, and shortness of breath in the past 3 months. The patient reported excessive weight loss of 10 lbs in the last month. On physical examination, her weight was 40.6 kg (89 lbs, 8.1 oz) and height was 153.4 cm, with a body mass index of 17.30 kg/m2. Other significant physical examination findings included scleral icterus, edema (4+ in bilateral lower extremities), and tachycardia. Abdominal examination was remarkable for the presence of distension and ascites, with tenderness in the left lower quadrant. Patient had a history of Crohn disease with several failed therapies and at presentation was on Cimzia®, an antitumor necrosis factor α (TNF-α) monoclonal antibody therapy, and prednisone. Initial laboratory results at the emergency department showed concentrations of sodium of 130 mmol/L, potassium of 3.4 mmol/L, chloride of 101 mmol/L, and albumin of 1.7 g/dL. She was then admitted to the intensive care unit (ICU)3 for intensive monitoring and correction of her electrolyte abnormalities. Further laboratory tests demonstrated hyperbilirubinemia (mainly conjugated bilirubin) and increased liver enzymes, including aspartate aminotransferase, alanine aminotransferase, and γ-glutamyltransferase, and increased prothrombin time and partial thromboplastin time (Table 1). Her …

Infliximab-Induced Aseptic Meningitis in a Patient with Crohnʼs Disease

Infliximab is an anti-tumor necrosis factor α (TNFα) monoclonal antibody that treats moderate-to-severe Crohn’s disease. In rare cases, infliximab has been associated with drug-induced aseptic meningitis. We present a 46-year-old woman with migraines and inflammatory Crohn’s colitis treated with intravenous infliximab and methotrexate. She developed nuchal rigidity, photophobia, and headache 2 days after each of her infliximab infusions, with symptom resolution 1 week post-infusion. Her exam, imaging, and cerebrospinal fluid analysis were consistent with drug-induced aseptic meningitis. She discontinued infliximab and started vedolizumab with continued remission of her Crohn’s disease.

Effects of tumor necrosis factor-α monoclonal antibody on nuclear factor-κB activation and inducible nitric oxide synthase expression in rats with silicotic fibrosis

Objective: To investigate the effects of tumor necrosis factor-α (TNF-α) monoclonal anti-body on nuclear factor-κB (NF-κB) activation and inducible nitric oxide synthase (iNOS) expression in rats with pulmonary fibrosis induced by silica dust. Methods: A total of 48 male Wistar rats were randomly divided into intervention group, silica dust exposure group, and control group, with 16 rats in each group. The rats in the intervention group were given intratracheal injection of 50 mg silicon dioxide dust once to establish a rat model and then treated with subcutaneously injected TNF-α monoclonal antibody 15 mg/kg for 5 consecutive days at 2-6 days after the establishment of the model. The rats in the silica dust exposure group were treated with the same method to establish the model and then given subcutaneous injection of the same volume of normal saline. The rats in the control group were given intratracheal and subcutaneous injection of normal saline. In both groups, 8 rats each were sacrificed at 7 and 14 days after the establishment of the model. Hematoxylin-eosin staining or Masson staining was used to observe morphological changes in lung tissue, ELISA was used to measure the serum level of TNF-α, IHC was used to measure the expression of NF-κBp65 in lung tissue, Western blot was used to measure the protein expression of I-κB in lung tissue, and RT-qPCR was used to measure the transcriptional level of iNOS mRNA in lung tissue. Results: Compared with the control group, the silica dust exposure group had significant increases in the lung inflammation score (3.375±1.061 and 2.500±0.535) , serum TNF-α level (86.405±20.494 and 77.064±11.829) , absorbance of cells with positive NF-κBp65 in lung tissue (0.297±0.05 and 0.287±0.039) , and mRNA expression of iNOS (12.906±0.590 and 12.600±0.517) at 7 and 14 days after dust exposure, a significant increase in pulmonary fibrosis score at 14 days (3.250±0.707) , and significant reductions in the protein expression of I-κB at 7 and 14 days (0.579±0.141 and 0.748±0.081) (P<0.05) . Compared with the silica dust exposure group, the intervention group had significant reductions in the lung inflammation score at 7 days (2.375±1.061) , pulmonary fibrosis score at 14 days (2.375±1.061) , serum level of TNF-α at 7 and 14 days (66.565±19.850 and 58.734±16.335) , absorbance of cells with positive NF-κBp65 in lung tissue at 7 and 14 days (0.248±0.028 and 0.238±0.027) , and mRNA expression of iNOS at 7 and 14 days (11.656±0.405 and 12.025±0.618) , as well as significant increases in the protein expression of I-κB at 7 and 14 days (0.802±0.165 and 0.888±0.144) (P<0.05) . Conclusion: TNF-α monoclonal antibody can inhibit the activation of the NF-κB signaling pathway and down-regulate the expression of iNOS, and thus exerts a certain protective effect on lung tissue in rats with pulmonary fibrosis induced by silica dust.

Infliximab Selectively Modulates the Circulating Blood Monocyte Repertoire in Crohn's Disease.

Infliximab (IFX), an anti-tumour necrosis factor alpha (TNFα) monoclonal antibody, provides clinical benefits in treating Crohn's disease (CD) but its mechanisms of action are not fully elucidated. This study investigated blood monocyte repertoires and the acute effects of IFX infusion on monocyte subset phenotype and function in IFX-treated patients with CD. Monocytes and monocyte subsets were enumerated and phenotypically characterized by multicolor flow cytometry in freshly isolated blood from healthy controls (n = 21) and patients with CD treated with (IFX, n = 24) and without (non-IFX, n = 20) IFX. For the IFX-CD group, blood was sampled immediately before (tough-IFX) and after (peak-IFX) infusion. Monocyte responses to lipopolysaccharide were analyzed by whole-blood intracellular cytokine staining. Non-IFX and IFX-CD patients had increased numbers of intermediate (CD14CD16) monocytes compared with healthy controls, whereas classical (CD14CD16) and nonclassical (CD14CD16) monocytes were numerically reduced in the IFX-CD group alone. In all groups, monocyte subsets expressed high surface levels of transmembrane (tm)TNFα. After IFX infusion, a significant reduction in monocyte numbers occurred. Post-IFX monocytopenia was proportionately greatest for classical and intermediate subsets, correlated with postinfusion IFX levels and was not associated with monocyte apoptosis. In contrast, lipopolysaccharide-induced production of TNFα and IL-12 by monocytes was significantly reduced in peak-IFX compared with trough-IFX blood samples. Actively managed CD is associated with monocyte repertoire skewing suggestive of chronic inflammatory stimulation. Infused IFX acutely targets monocytes, likely by binding to tmTNFα, resulting in a non-apoptosis-related decline in circulating monocyte numbers and blunting of the inflammatory response of monocytes remaining in the blood.

A comparative Study of Immunohistochemical Expression of Tumor Necrosis Factor-Alpha, Interleukin-6and Vascular Endothelial Growth Factor in Giant Cell Granuloma of the Jaws and Giant Cell Tumor of Long Bones

Background: Central giant cell granuloma (CGCG) and peripheral giant cell granuloma(PGCG) are pathological conditions of the jaws that share the same microscopic features, but differ clinically in terms of their behavior. While the giant cell tumor (GCT) of long bones is a rare benign neoplasm, tend to affect femur and tubular bone, characterized by local aggressiveness, high recur- rence rates and metastasis to the lung. Objectives: To evaluate, compare and correlate the expression of TNF-α, IL-6 and VEGF in peripheral and central giant cell granu - loma of the jaw and giant cell tumor of long bones. Methods: A total of 60 retrospective formalin- fixed, paraffine-embeded specimens of giant cell lesions of the jaws and long bones, where included in this study. An immunohistochemical staining with TNF-α , IL-6 and VEGF monoclonal antibodies were performed. Results: TNF-α, IL-6 and VEGF were expressedin all lesions. The PGCG comparedto the CGCG and GCT showed significantly in - creased expression of TNF-α and decreased expression of VEGF by the stromal cells..GCT showed increased expression of VEGF by M NGCs and stromal cells .There is a non significant difference between CGCG and GCT regarding the expression of all three cytokines. Conclusions: The present study confirmed the usefulness TNF-α,IL-6 and VEGF in evaluating osteoclastogenesis. The results of this study proved that the biological activity of TNF-α, IL-6 and VEGF was comparable between the central giant cell granuloma and giant cell tumors, supporting the observations that these two lesions are the same entity and have the same biological behavior.

RhTNFR: Fc Suppresses the Development of Endometriosis in a Mouse Model by Downregulating Cell Proliferation and Invasiveness.

Tumor necrosis factor α (TNF-α), a proinflammatory cytokine, may play an important role in the pathogenesis of endometriosis; therefore, TNF-α inhibitors potentially have an effect on endometriosis. To investigate the effect of anti-TNF-α treatment on endometriosis, 2 TNF-α inhibitors: recombinant human TNF receptor: Fc fusion protein (rhTNFR: Fc) and TNF-α monoclonal antibody (TNF-α mAb) were used to treat human eutopic endometrial stromal cells (hESCs), and the effects on cell survival, cell cycle, and invasiveness were compared. It was found that rhTNFR: Fc suppressed the TNF-α-induced hESC survival and invasiveness but not TNF-α mAb. Recombinant human TNF receptor: Fc fusion protein decreased the S phase of hESC compared with the TNF-α-treated group. Then, we used a surgically induced mouse model of endometriosis to study the effect of rhTNFR: Fc treatment in vivo. The fluorescence intensity and the size of implanted endometriotic lesions in the mouse model were decreased by rhTNFR: Fc. In conclusion, rhTNFR: Fc suppresses hESC survival and invasiveness and decreases the fluorescence intensity and implant size in the mouse model of endometriosis.

Reduced numbers of mucosal DRint macrophages and increased numbers of CD103+ dendritic cells during anti-TNF-α treatment in patients with Crohn’s disease

Objective Anti-TNF-α treatment constitutes a mainstay in the treatment of Crohn’s disease (CD), but its mechanisms of action are not fully understood. We aimed to investigate the effects of adalimumab, a human monoclonal TNF-α antibody, on macrophage (MQ) and dendritic cell (DC) subsets in mucosal biopsies and peripheral blood. Material and methods Intestinal biopsies and blood samples were obtained from 12 different CD patients both before and 4 weeks after the initiation of the induction of adalimumab treatment. Endoscopic disease activity was estimated by the Simple Endoscopic Score for Crohn’s Disease. Biopsies were obtained from inflamed and non-inflamed areas. The numbers of lamina propria CD14 + DRint and CD14 + DRhi MQs, CD141+, CD141− and CD103+ DCs subsets, and circulating monocytes and DCs were analyzed using flow cytometry. Results At baseline, we observed higher numbers of DRint MQs and lower numbers of CD103+ DCs in inflamed versus non-inflamed mucosa [843 vs. 391/105 lamina propria mononuclear cells (LPMCs) (p < 0.05) and 9 vs. 19 × 105 LPMCs (p = 0.01), respectively]. After four weeks of adalimumab treatment, the numbers of DRint MQs decreased [843 to 379/105 LPMCs (p = 0.03)], whereas the numbers of CD103+ DCs increased [9–20 × 105 LPMCs (p = 0.003)] compared with baseline. In peripheral blood, no alterations were observed in monocyte or DC numbers between baseline and week 4. Conclusions In CD, mucosal inflammation is associated with high numbers of DRint MQs and low numbers of CD103+ DCs. This composition of intestinal myeloid subsets is reversed by anti-TNF-α treatment. These results suggest that DRint MQs play a pivotal role in CD inflammation.

Systemic administration of an anti-tumor necrosis factor-alpha monoclonal antibody protects against endotoxin-induced uveitis in rats.

Objective:This study was to evaluate the effect of systemic injection of an anti-tumor necrosis factor alpha (TNF-α) monoclonal antibody (mAb) on endotoxin-induced uveitis (EIU).Materials and Methods:Fifty-six male Wistar rats (6–8 weeks old) were randomly divided into three groups: EIU, anti-TNF-α mAb + EIU, and control. EIU was induced by injecting Escherichia coli O55:B5 lipopolysaccharide (LPS) into the hind footpad of the rats (150 μg/rat). The anti-TNF-α mAb (1 μg/kg) was administrated 30 min before LPS injection through one-time intravenous injection. The onset time and peak time of EIU were recorded. The serum and aqueous humor (AH) TNF-α, interleukin (IL)-6, and IL-10 levels were measured by ELISA at 4, 24, and 72 h post-LPS injection. Clinical manifestations of EIU and eye histopathology were scored.Results:Compared with the EIU rats, anti-TNF-α mAb + EIU rats showed significantly delayed onset of uveitis (t = 7.41, P < 0.001), lower clinical scores and histopathological grades (t = 3.18/2.22, P < 0.001), reduced levels of TNF-α (F = 15.06/59.43, P < 0.001) and IL-6 (F = 99.63/14.92, P < 0.001), and increased levels of IL-10 (F = 24.94/8.99, P < 0.001) in the serum and AH. AH TNF-α, serum IL-6, and AH IL-6 levels are positively correlated, whereas serum IL-10 levels were negatively correlated with EIU activity.Conclusion:Antagonizing TNF-α by system injection of the anti-TNF-α mAb protects against EIU in rats. Blocking TNF-α signaling could be a useful strategy for managing uveitis.

IL-1β, in contrast to TNFα, is pivotal in blood-induced cartilage damage and is a potential target for therapy

AbstractJoint bleeding after (sports) trauma, after major joint surgery, or as seen in hemophilia in general leads to arthropathy. Joint degeneration is considered to result from the direct effects of blood components on cartilage and indirectly from synovial inflammation. Blood-provided proinflammatory cytokines trigger chondrocytes and induce the production of cartilage-degrading proteases. In the presence of erythrocyte-derived iron, cytokines stimulate radical formation in the vicinity of chondrocytes inducing apoptosis. To unravel the role of interleukin (IL) 1β and tumor necrosis factor (TNF) α in the pathogenesis of this blood-induced cartilage damage, the effect of antagonizing these cytokines was examined in human in vitro cultures. Addition of recombinant human IL-1β monoclonal antibody or IL-1 receptor antagonist resulted in a dose- and time-dependent protection of cartilage from blood-induced damage. In higher concentrations, almost complete normalization of cartilage matrix proteoglycan turnover was achieved. This was accompanied by a reduction in IL-1β and IL-6 production in whole blood cultures, whereas TNFα production remained unaffected. Interestingly, addition of a TNFα monoclonal antibody, although demonstrated to inhibit the direct (transient) effects of TNFα on cartilage, exhibited no effect on blood-induced (prolonged) cartilage damage. It is demonstrated that IL-1β is crucial in the development of blood-induced joint damage, whereas TNFα is not. This hierarchical position of IL-1β in blood-induced joint damage warrants studies on targeting IL-1β to potentially prevent joint degeneration after a joint bleed.

TNF-α produced by SEC2 mutant (SAM-3)-activated human T cells induces apoptosis of HepG2 cells.

Staphylococcal enterotoxins C2 (SEC2) is a classical model of superantigens (SAg), which has the powerful ability to activate T cells as well as induce massive cytokine production. This property makes SEC2 and its mutants well concerned as a potential new immune-regulatory agent for cancer therapy. We previously constructed a SEC2 mutant named SAM-3, which had prominently antitumor activity in BALB/c mice model. But, the underlying molecular mechanism for stimulation of human peripheral blood mononuclear cells (PBMCs) and antitumor effect on human tumor cells induced by SAM-3 is not clear. Here, we showed that SAM-3 could activate human TCR Vβ 12, 13A, 14, 15, 17, and 20 CD8(+) subgroup T cells, which secreted the cytokines IL-2, IFN-γ, and TNF-α, and exhibit stimulation activity in a dose-dependent manner. TNF-α secreted from activated T cells could induce apoptosis and G1-phase arrest and lead to the antitumor effect in HepG2 cells. Meanwhile, SAM-3 upregulated the expression of tumor necrosis factor receptor 1 (TNFR1) mRNA and activity of caspase-3 and caspase-8. We also found that the antitumor activity and activity of caspase-3 and caspase-8 were decreased when the neutralizing TNF-α monoclonal antibody presented. These data suggest that TNF-α secreted by SAM-3-activated T cells is an important factor in inducing apoptosis in HepG2 cells.

Golimumab in the treatment of psoriatic arthritis: efficacy and safety

Tumor necrosis factor-α (TNF-α) holds a central position in the pathogenesis of autoimmune inflammatory diseases of the locomotor apparatus. A separate class of drugs, namely, TNF-α inhibitors, that are effective against multicomponent diseases, such as psoriatic arthritis (PsA), is now available to physicians. The paper reviews the results of clinical trials of the TNF-α inhibitor golimumab, a human TNF-α monoclonal antibody. Golimumab exerts a positive effect on all manifestations of PsA: arthritis, psoriatic skin and nail lesions, dactylitis, enthesitis, and quality of life. The drug is noted for its convenient route of administration – its standard dose is 50 mg injected subcutaneously once a month and for its low molecular immunogenicity. Recent data suggest that golimumab is an effective drug with a safety profile similar to that of the entire class of TNF-α inhibitors.

Nemesis of neglected neurosarcoidosis.

Nemesis of neglected neurosarcoidosis.

Role of the glucocorticoid receptor in the recurrence of primary nephrotic syndrome.

The present study aimed to investigate the changes in the expression levels of the glucocorticoid receptor (GR) and its subtypes in patients with recurrent renal syndrome. In addition, the effects of tumour necrosis factor α (TNF-α) and a TNF-α monoclonal antibody on these receptors in peripheral blood mononuclear cells (PBMCs) isolated from the patients was analysed. Furthermore, a new treatment method for recurrent renal syndrome was explored. The serum levels of TNF-α in the normal (A), stable renal syndrome (B) and renal syndrome recurrence (C) groups of patients were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression levels of GR, GRα and GRβ were determined by ELISA, western blot analysis and quantitative polymerase chain reaction in PBMC cultures from the three groups in the absence of intervention (blank control) and following stimulation with methylprednisolone, TNF-α and/or TNF-α monoclonal antibody. Group C exhibited higher expression levels of TNF-α and GRβ but a lower level of GRα expression (P<0.05) compared with the other groups. Regardless of methylprednisolone intervention, the expression levels of GR and GRβ in the three groups following stimulation by TNF-α were significantly higher compared with those in the respective blank control, whereas in group C, the GRα expression levels following TNF-α treatment were lower compared with those in the control group (P<0.05). The treatment of group C with TNF-α monoclonal antibodies resulted in higher GRα expression but lower GRβ expression compared with those in the blank control (P<0.05). The change in the ratios of the GR subtypes may be associated with renal syndrome recurrence. TNF-α may be involved in renal syndrome relapse by changing the levels of GR as well as the proportion of the GR subtypes. TNF-α monoclonal antibodies may mitigate the changes in the ratios of these subtypes.

Effects of monocyte-endothelium interactions on the expression of type IV collagenases in monocytes.

The adhesion of monocytes to endothelial cells is one of the early stages in the development of atherosclerosis. The expression of type IV collagenases, which include matrix metalloproteinase (MMP)-2 and MMP-9, in monocytes is hypothesized to play an important role in monocyte infiltration and transformation into foam cells. The aim of the present study was to examine the effects of monocyte-endothelium interactions on the expression levels of type IV collagenases and their specific inhibitors in monocytes, and to investigate the roles of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in this process. Monocytes were single-cultured or co-cultured with endothelial cells. The expression of the type IV collagenases, MMP-2 and MMP-9, and their specific inhibitors, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2, in monocytes was determined by immunohistochemistry followed by image analysis. The expression levels of MMP-2 and MMP-9 were found to be low in the single-culture monocytes, but increased significantly when the monocytes and endothelial cells were co-cultured. However, treatment with monoclonal TNF-α or IL-1β antibodies partially inhibited the upregulated expression of MMP-2 and MMP-9 in the co-cultured monocytes. Expression of TIMP-1 and TIMP-2 was observed in the single monocyte culture, and a small increase in the expression levels was observed when the monocytes were co-cultured with endothelial cells. Therefore, monocyte-endothlium interactions were shown to increase the expression of type IV collagenases in monocytes, resulting in the loss of balance between MMP-2 and -9 with TIMP-1 and -2. In addition, TNF-α and IL-1β were demonstrated to play important roles in this process.

Comparison of the efficacies of seton drainage combined with different drugs in the treatment of Crohn's disease complicated with anal fistula

Objective To compare the efficacies of seton drainage combined with anti TNF-α monoclonal antibody or immunosuppressants in the treatment of Crohn's disease complicated with anal fistula.Methods The clinical data of 37 patients with Crohn's disease complicated with anal fistula who were admitted to the Sixth Affiliated Hospital of Sun Yat-Sen University from June 2007 to February 2014 were retrospectively analyzed.Seventeen patients were treated by seton drainage combined with anti TNF-α monoclonal antibody (anti TNF-α monoclonal antibody group) and 20 were treated by seton drainage combined with immunosuppressants (immunosuppressant group).The ratios of patients with healed and unhealed fistula,time for fistula heal,ratio of anal fistula recurrence,time for anal fistula recurrence and ratio of reoperation for anal fistula between the 2 groups were compared.Patients were followed up via out-patient examination and phone call till May 2014.The measurement data and the count data were analyzed using the t test and chi-square test,respectively.Results One patient in the anti TNF-α monoclonal antibody group was treated by adalimumab,and the other 16 patients were treated by infliximab.Twenty patients in the immunosuppressant group were treated by azathioprine.All the patients were followed up for a mean time of 19 months (range,3-50 months).The ratios of patients with healed and unhealed fistula,recurrence of anal fistula,reoperation for anal fistula recurrence,time for fistula heal and time for anal fistula recurrence were 8/17,7/17,2/17,2/2,(2.5 ± 2.1)months and (4.0 ± 2.8)months in the anti TNF-α monoclonal antibody group,and 12/20,3/20,5/20,3/5,(3.6 ± 3.5) months and (9.0 ± 8.2) months in the immunosuppressant group,with no significant difference between the 2 groups (x2=0.620,2.137,0.364,0.018,t =-1.035,-0.619,P ＞ 0.05).Conclusions Seton drainage is effective for the treatment of Crohn's disease complicated with anal fistula.The efficacies of seton drainage combined with anti TNF-α monoclonal antibody or immunosuppressant are comparative. Key words: Crohn's disease ; Anal fistula; Seton drainage ; Anti TNF-α monoclonal antibody; Immunosuppressant

Dual effects of tumor necrosis factor alpha on myocardial injury following prolonged hypoperfusion of the heart

Objective: To examine the dose response of TNFα in an ex vivo rat model of myocardial ischemia reperfusion. Methods and Results: Seventy-two rat hearts were mounted on Langendorff apparatus and perfused with oxygenated Krebs-Henseleit solutions. Ischemia was induced by reducing the perfusate flow rate. During reperfusion, incremental doses of recombinant TNFα were infused as a part of perfusate. TNFα was blocked with monoclonal TNFα antibody. Myocardial function was measured by dP/dT and relaxation time (IVRT). Cellular injury was assessed by released myoglobin and tissue concentration of malondialdehyde activity of the heart homogenates. Baseline +dP/dT was 1645 ± 125 mmHg/sec, –dP/dT was 945 ± 73 mmHg/sec and IVRT was 65 ± 5 msec. At the conclusion of reperfusion period, lower doses of TNFα increased +dP/dT and lowered IVRT. In contrast, the higher doses of TNFα decreased +dP/dT and prolonged IVRT. Pretreating the hearts with monoclonal TNFα antibody completely abolished the effects of TNFα on myocardial contractility and relaxation comparable to ischemia controls. Conclusion: Low dose TNFα improved myocardial function and decreased resultant cellular injury while high dose TNFα decreased myocardial function and increased myocardial injury following ischemia and reperfusion.