Anti-tumor Necrosis Factor-alpha Antibody Research Articles

Evaluation of a TGN1412 analogue using in vitro assays and two immune humanized mouse models

Cytokine release syndrome (CRS) is a serious and potentially life-threatening complication typically associated with biological drug products. Pre-clinical testing in vitro and in vivo studies using non-human primates had failed to reliably predict CRS. To determine if bone marrow-thymus-liver (BLT) humanized mice with a fully engrafted human immune system or a CD34-humanized mouse model could predict CRS, we tested an anti-CD28 monoclonal antibody (mAb) similar to TGN1412. This TGN1412 analogue (TGN1412A) was initially tested in vitro and found to produce significant dose-dependent increases in cytokine production. For in vivo studies, adalimumab, an anti-tumor necrosis factor-alpha antibody known not to cause CRS, served as a negative control. We evaluated immune cell activation and cytokine expression in three independent experiments. In BLT humanized mice, significant increases in levels of human cytokines were identified in animals treated with anti-CD28 mAb. As expected, CD28+ cell detection was strongly reduced in the anti-CD28 treated group. Increased T cell activation was also observed. The control group did not show reductions in CD28+ T-cells and did not experience increased cytokine levels. Responses by CD34-humanized mice showed no significant differences between adalimumab and anti-CD28 treatment at doses used to test BLT-humanized mice. These results suggest that the TGN1412A produces similar results in vitro to the original TGN1412 monoclonal antibody. The BLT immune humanized mice but not the CD34 humanized mice produce both robust and specific cytokine responses and may represent a pre-clinical model to identify CRS.

Safety and efficacy of infliximab in the treatment of refractory uveoretinitis in Beh\xe7et\u2019s disease: a large-scale, long-term postmarketing surveillance in Japan

BackgroundInfliximab, an anti-tumor necrosis factor-alpha antibody, has been reported to have excellent efficacy for refractory uveoretinitis in Behçet’s disease (RUBD), and was approved for this indication in Japan. However, the long-term safety profile and efficacy in real-world clinical settings in patients with RUBD have not been fully clarified. The BRIGHT study, a prospective, large-scale, long-term postmarketing surveillance (PMS) study, was conducted to investigate the long-term safety and efficacy of infliximab in Japanese patients with RUBD.MethodsAll patients with RUBD who started infliximab treatment between January 2007 and January 2010 were enrolled. Safety was evaluated every 6 months for up to 24 months after initiation of therapy in 656 patients, and efficacy was evaluated in 650 patients. Patient characteristics were compared using the chi-square or Fisher’s exact test. The frequency of ocular attacks before and after infliximab treatment was compared using the Wilcoxon signed-rank test. Independent associated factors for safety or efficacy were identified using multiple logistic regression analysis. A two-sided p value <0.05 was considered significant.ResultsAmong the 656 patients evaluated for safety, 555 (84.6%) completed the 24-month study period. The incidence of adverse drug reactions (ADRs) and serious ADRs were 32.32% and 6.10%, respectively, and the safety profile was comparable to that of Japanese PMS of infliximab for other diseases. The most common ADRs and serious ADRs were infections (11.89% and 3.66%). Tuberculosis was reported in two patients, and Pneumocystis jirovecii in one. Identified independent associated factors for infections were comorbid respiratory disease, history of allergic disease, and concomitant use of glucocorticoids. Although infusion reactions were observed in 11.13% of patients, most were non-serious. The response rate at 24 months by physician global assessment was 80.7%. Median frequency of ocular attacks per 6 months significantly decreased compared with that before infliximab treatment (2.0 to 0.0), and corrected visual acuity was maintained during the study.ConclusionsInfliximab treatment had good tolerability and efficacy in Japanese patients with RUBD in this large-scale, long-term PMS. Infliximab treatment seemed to be a good treatment option for RUBD in real-world clinical settings.Trial registrationUMIN Clinical Trials Registry, UMIN000027733. Retrospectively registered on 6 June 2017.

Production and Characterization of a New Variant of an Anti-Tnfα Antibody with Improved Affinity and Potency

Rapid growth in the therapeutic antibody market leads to a drastic development of antibody engineering to optimize biophysical properties. One of the main focuses of biopharmaceutical researches was the expansion of different approaches for affinity maturation of antibodies to enhance biological activity. Adalimumab (D2E7) is an anti-tumor necrosis factor alpha (TNFα) antibody used in the treatment of some autoimmune disorders like rheumatoid arthritis, psoriasis, etc. In this study, by engineering complementary determining regions (CDRs) of D2E7 antibody, we produced a new variant of the antibody with improved affinity and potency to TNFα. We designed four D2E7 mutants that harbored a single point mutation in their CDRs. The native antibody, lc-A94K, lc-A50Y, lc-A33R, and lc-A92R mutant models were transiently produced and characterized. Data showed mutation of ALA to LYS in CDR3 of D2E7 antibody generated new hydrogen bonds with TNFα, thus the lc-A94K model revealed significantly higher binding activity (EC50) and kinetic affinity (KD) to its antigen, in comparison to the wild antibody. Moreover, this model was found to have significantly stronger biological activity (IC50) to activate antibody-dependent cell-mediated cytotoxicity in the mouse fibroblast L929 cells. Secondary structure analysis demonstrated the mutation has no inappropriate conformational impact on the beta and alpha structures of lc-A94K antibody. In conclusion, our study revealed that it is possible to manipulate antibody’s CDRs to increase affinity and potency by single point mutation and also preserve basic structure of the original antibody during CDRs engineering to avoid adverse effects of CDRs mutations on specificity and stability.

Perianal pediatric Crohn disease is associated with a distinct phenotype and greater inflammatory burden

Growth deceleration and impaired pubertal growth spurt are common concerns in patients with early onset inflammatory bowel disease (IBD) (1-2). Previous studies confirmed the efficacy and safety of infliximab, an anti-tumor necrosis factor alpha (TNF-α) antibody, in achieving clinical remission in luminal Crohn’s disease (CD) and in facilitating linear growth (3). The present study describes in depth the specific characteristics of children with perianal CD (pCD), a sub-category of CD patients that appears to exhibit a higher level of inflammation. Previous studies reported a higher rate of intestinal resection in patients with pCD. As the authors state, the lack of this association in the present cohort could be linked to the currently recommended prompt start of anti-TNFa agents in the presence of perianal disease, that alters the natural history of the disease. This study reports reduced height Z-scores in children with pCD. It should be noted that height data were not correlated to the target height. Similarly, no data about growth velocity, pubertal stage and/or bone age of patients were available. For these reasons, it appears interesting to investigate in more detail the growth aspects in this sub-category of patients with IBD.

Allergic and Immunologic Perspectives of Inflammatory Bowel Disease.

Inflammatory bowel disease (IBD) is a chronic immune-mediated inflammatory condition primarily involving the gastrointestinal tract. It includes Crohn's disease (CD), ulcerative colitis (UC), and a less common phenotype-indeterminate colitis. It is thought to result from a complex interplay of environmental, microbial, and host factors including genetic factors, although the exact mechanism is not known. Dietary factors have been shown to play a role in the pathogenesis of IBD and can potentially alter the intestinal microbiota as well as disrupt the immune function in the gut. CD is characterized by transmural inflammation, sometimes associated with granulomatous lesions, and involves the entire gastrointestinal tract but often spares the rectum. UC is characterized by mucosal inflammation typically confined to the colon and rectum. Although IBD is mostly seen in western world, recent data suggests that the incidence and prevalence are increasing worldwide. Enteral nutrition has been shown to be effective in inducing remission in pediatric population with CD; however, there is mixed data in adult population. Nutritional deficiencies such as vitamin D and zinc deficiency are often noted in IBD patients. Several extraintestinal manifestations are noted in patients with IBD. Some of them parallel with the disease activity and others are independent of the disease course. Assessment of IBD disease activity clinically, radiologically, if indicated, biochemically and endoscopically is important to guide therapy in IBD. To ensure comprehensive care, it is important to assess associated conditions such as nutritional and psychological well-being, as well as age appropriate health maintenance status prior to starting treatment for IBD. Several biologic agents including anti-tumor necrosis factor alpha (anti-TNF-α) drugs, anti-integrins, and antibodies to the p40 subunit of IL12/23 are approved for induction and maintenance of remission of IBD. Steroids are also often used for induction. Anti-metabolites and thiopurines are also useful either as monotherapy or in combination regimens. Potential side effects of anti-TNF-α drugs such as serious infections, malignancy, worsening of heart failure, and infusion-related reactions should be considered prior to starting these drugs. Anti-TNF-α drugs with or without immunomodulators (azathioprine, 6-mercaptopurine, methotrexate) are often used for the induction and maintenance of remission. Treating to target of endoscopic and clinical remission provides the best long-term outcomes. Our knowledge and understanding of IBD has grown significantly. However, there are several unanswered questions on pathogenesis, disease behavior, and drivers of inflammation in various patient subgroups which require further research.

Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk.

BackgroundSubarachnoid hemorrhage (SAH) can induce apoptosis in many regions of the brain including the cortex and hippocampus. However, few studies have focused on apoptosis in the hypothalamus after SAH. Although some antiapoptotic strategies have been developed for SAH, such as anti-tumor necrosis factor-alpha (TNF-α) antibody, the molecular mechanisms underlying this condition have yet to be elucidated. Therefore, the purpose of this study was to evaluate whether SAH could induce apoptosis in the hypothalamus and identify the potential molecular mechanisms underlying the actions of anti-TNF-α antibody, as a therapeutic regimen, upon apoptosis.Materials and methodsSAH was induced in a rat model. Thirty minutes prior to SAH, anti-TNF-α antibody or U0126, an extracellular signal-regulated kinase (Erk) inhibitor, was microinjected into the left lateral cerebral ventricle. In addition, phorbol-12-myristate-13-acetate was injected intraperitoneally immediately after the anti-TNF-α antibody microinjection. Then, real-time polymerase chain reaction, Western blotting and immunohistochemistry were used to detect the expression of caspase-3, bax, bcl-2, phosphorylated Erk (p-Erk) and Erk. Finally, anxiety-like behavior was identified by using open field.ResultsLevels of caspase-3, bax and bcl-2, all showed a temporary rise after SAH in the hypothalamus, indicating the induction of apoptosis in this brain region. Interestingly, we found that the microinjection of anti-TNF-α antibody could selectively block the elevated levels of bax, suggesting the potential role of anti-TNF-α antibody in the inhibition of SAH-induced apoptosis in the hypothalamus. Moreover, we found that Erk activation was necessary for apoptosis after SAH and that the microinfusion of anti-TNF-α antibody could inhibit apoptosis by suppressing the increase of p-Erk in the hypothalamus. Finally, our data indicated that the infusion of anti-TNF-α antibody could improve anxiety-like behavior.ConclusionCollectively, our data demonstrate that anti-TNF-α antibody attenuates apoptosis in the hypothalamus by inhibiting the activation of Erk, which plays an important role in the treatment of SAH.

Comparison of Treatment Guidelines for Ulcerative Colitis: Role of Biologics

Ulcerative colitis (UC) is a chronic inflammatory condition of the colon, characterized by diffuse mucosal inflammation with bloody diarrhea. The therapeutic goals in UC have evolved from symptomatic relief to deep remission, including sustained steroid-free clinical remission, mucosal healing, and ideally histologic remission. The management of UC has significantly been improved by advance of biologic agents including anti-tumor necrosis factor alpha (TNF-α) antibodies. Anti-TNF-α agent have progressively been introduced earlier in treatment algorithms for UC in order to minimize steroid exposure and dependence and to maximize disease control and quality of life. We reviewed the therapeutic recommendations of biologic agents for UC in the Korean and European Crohn's and Colitis Organization guidelines.

Assessment of Antibody-based Drugs Effects on Murine Bone Marrow and Peritoneal Macrophage Activation.

Macrophages are phagocytic innate immune cells, which initiate immune responses to pathogens and contribute to healing and tissue restitution. Macrophages are equally important in turning off inflammatory responses. We have shown that macrophages stimulated with intravenous immunoglobulin (IVIg) can produce high amounts of the anti-inflammatory cytokine, interleukin 10 (IL-10), and low levels of pro-inflammatory cytokines in response to bacterial lipopolysaccharides (LPS). IVIg is a polyvalent antibody, primarily immunoglobulin Gs (IgGs), pooled from the plasma of more than 1,000 blood donors. It is used to supplement antibodies in patients with immune deficiencies or to suppress immune responses in patients with autoimmune or inflammatory conditions. Infliximab, a therapeutic anti-tumor necrosis factor alpha (TNFα) antibody, has also been shown to activate macrophages to produce IL-10 in response to inflammatory stimuli. IVIg and other antibody-based biologics can be tested to determine their effects on macrophage activation. This paper describes methods for derivation, stimulation, and assessment of murine bone marrow macrophages activated by antibodies in vitro and murine peritoneal macrophages activated with antibodies in vivo. Finally, we demonstrate the use of western blotting to determine the contribution of specific cell signaling pathways to anti-inflammatory macrophage activity. These protocols can be used with genetically modified mice, to determine the effect of a specific protein(s) on anti-inflammatory macrophage activation. These techniques can also be used to assess whether specific biologics may act by changing macrophages to an IL-10-producing anti-inflammatory activation state that reduces inflammatory responses in vivo. This can provide information on the role of macrophage activation in the efficacy of biologics during disease models in mice, and provide insight into a potential new mechanism of action in people. Conversely, this may caution against the use of specific antibody-based biologics to treat infectious disease, particularly if macrophages play an important role in host defense against that infection.

Stat3 as a potential therapeutic target for rheumatoid arthritis

Rheumatoid arthritis (RA) is a multi-factorial disease characterized by chronic inflammation and destruction of multiple joints. To date, various biologic treatments for RA such as anti-tumor necrosis factor alpha antibodies have been developed; however, mechanisms underlying RA development remain unclear and targeted therapy for this condition has not been established. Here, we provide evidence that signal transducer and activator of transcription 3 (Stat3) promotes inflammation and joint erosion in a mouse model of arthritis. Stat3 global KO mice show early embryonic lethality; thus, we generated viable Stat3 conditional knockout adult mice and found that they were significantly resistant to collagen-induced arthritis (CIA), the most common RA model, compared with controls. We then used an in vitro culture system to screen ninety-six existing drugs to select Stat3 inhibitors and selected five candidate inhibitors. Among them, three significantly inhibited development of arthritis and joint erosion in CIA wild-type mice. These findings suggest that Stat3 inhibitors may serve as promising drugs for RA therapy.

Inflammatory Bowel Disease: Joint Management in Gastroenterology and Dermatology

Inflammatory bowel disease (IBD) is a complex entity that includes Crohn disease and ulcerative colitis. It is characterized by a chronic proinflammatory state of varying intensity that often leads to considerable morbidity. In the last decade, several therapeutic targets have been identified that are susceptible to the use of biological agents, including anti-tumor necrosis factor alpha antibodies, which are associated with paradoxical psoriasiform reactions in 5% of patients. Decision-making in the management of these cases requires close collaboration between the dermatologist and gastroenterologist. Inflammatory bowel disease is also associated with various other dermatologic and rheumatologic manifestations, and presents a genetic and pathogenic association with psoriasis that justifies both the interdisciplinary approach to these patients and the present review.

An Update on Treatment of Pediatric Chronic Non-Infectious Uveitis.

There are no standardized treatment protocols for pediatric non-infectious uveitis. Topical corticosteroids are the typical first-line agent, although systemic corticosteroids are used in intermediate, posterior and panuveitic uveitis. Corticosteroids are not considered to be long-term therapy due to potential ocular and systemic side effects. In children with severe and/or refractory uveitis, timely management with higher dose disease-modifying antirheumatic drugs (DMARDs) and biologic agents is important. Increased doses earlier in the disease course may lead to improved disease control and better visual outcomes. In general, methotrexate is the usual first-line steroid-sparing agent and given as a subcutaneous weekly injection at >0.5 mg/kg/dose or 10-15 mg/m2 due to better bioavailability. Other DMARDs, for instance mycophenolate, azathioprine, and cyclosporine are less common treatments for pediatric uveitis. Anti-tumor necrosis factor-alpha agents, primarily infliximab and adalimumab are used as second line agents in children refractory to methotrexate, or as first-line treatment in those with severe complicated disease at presentation. Infliximab may be given at a minimum of 7.5 mg/kg/dose every 4 weeks after loading doses, up to 20 mg/kg/dose. Adalimumab may be given up to 20 or 40 mg weekly. In children who fail anti-tumor necrosis factor-alpha agents, develop anti-tumor necrosis factor-alpha antibodies, experience adverse effects, or have difficulty with tolerance, there is less data available regarding subsequent treatment. Promising results have been noted with tocilizumab infusions every 2-4 weeks, abatacept monthly infusions and rituximab.

P215 Clinical features of chronic enteropathy associated with SLCO2A1 gene (CEAS)

Background: Chronic enteropathy associated with SLCO2A1 gene (CEAS) is a rare hereditary disease caused by mutations in SLCO2A1 gene, encoding a prostaglandin transporter. CEAS resembles Crohn's disease (CD) in presenting multiple small intestinal ulcers. However, because immunosuppressive therapies such as corticosteroid and anti-tumor necrosis factor-alpha antibody therapies are ineffective for CEAS in contrast to CD, recognition and precise diagnosis of CEAS are critical to avoid unnecessary therapies. Methods: Forty-one Japanese patients (10 male and 31 female) were diagnosed with CEAS by clinical findings and genetic analysis during a period of 2012 to 2016 and they were enrolled in this study. To identify the clinical features of CEAS, we reviewed the clinical information for the patients. In addition, because SLCO2A1 gene has also been reported as a causative gene of primary hypertrophic osteoarthropathy (PHO; OMIM 614441), we investigated whether CEAS patients had clinical manifestations of PHO. Results: We found recessive SLCO2A1 mutations located at 11 sites. Among the identified SLCO2A1 mutations, a splice-site mutation of intron 7 (c.940+1G>A) was the most frequent, and 17 of the 41 patients were homozygous for this mutation (41%). Median age at onset was 17 (1 to 69) and parental consanguinity was present in 12 patients (29%). Anemia and abdominal pain were present in 40 (98%) and 16 (39%) patients, respectively. On the other hand, there were little inflammatory findings in blood tests (median CRP 0.16mg/dl). During the course of the disease, 27 patients (66%) had undergone one or more surgeries such as partial resection of the small intestine. The most frequently involved gastrointestinal site was ileum (98%), but there was no patient with ulcerations in the terminal ileum. The stomach and duodenum were affected in 11 (27%) and 18 (44%) patients, respectively. Although no patients had clinical manifestations of PHO that required treatment, mild digital clubbing or periostosis was present in 12 patients (29%). Moreover, 4 male patients fulfilled the major clinical criteria for PHO, having digital clubbing, periostosis, and pachydermia. Conclusions: Although CEAS mimics CD, some clinical features of CEAS are different from those of CD, such as low inflammatory findings, terminal ileum-sparing ulcerations and extraintestinal manifestations. Genetic analysis should be considered if CEAS is clinically suspected.

Inhibiting tumor necrosis factor-alpha diminishes desmoplasia and inflammation to overcome chemoresistance in pancreatic ductal adenocarcinoma.

BackgroundPancreatic ductal adenocarcinoma (PDAC) is one of the most common cancer death reasons. Anti-tumor necrosis factor-alpha (TNF-α) antibodies have shown promising effects in PDAC pre-clinical models. However, the prognostic values of TNF-α, underlying mechanisms by which anti-TNF-α treatments inhibit PDAC, and potential synergistic effects of anti-TNF-α treatments with chemotherapy are still unclear.Results and MethodsTo identify the targeting values of TNF-α in PDAC, we measured TNF-α expression in different stages of PDAC initiation and evaluated its prognostic significance in a pancreatic cancer cohort. We found that TNF-α expression elevated in PDAC initiation process, and high expression of TNF-α was an independent prognostic marker of poor survival. We further evaluated anti-tumor effects of anti-TNF-α treatments in PDAC. Anti-TNF-α treatments resulted in decreased cell viability in both PDAC tumor cells and pancreatic satellite cells in similar dose in vitro. In vivo, anti-TNF-α treatments showed effects in reducing desmoplasia and the tumor promoting inflammatory microenvironment in PDAC. Combination of anti-TNF-α treatments with chemotherapy partly overcame chemoresistance of PDAC tumor cells and prolonged the survival of PDAC mouse model.ConclusionsIn conclusion, our findings indicated that TNF-α in PDAC can be a prognostic and therapeutic target. Inhibition of TNF-α synergized with chemotherapy in PDAC resulted in better pre-clinical responses via killing tumor cells as well as diminishing desmoplasia and inflammation in PDAC tumor stroma.

Golimumab Use in Bio-experienced Ulcerative Colitis Patients

Introduction: Golimumab is a human anti-tumor necrosis factor alpha (TNF- α) antibody approved for the treatment of moderate to severe ulcerative colitis (UC) in adults. It was found to be more efficacious compared to placebo in bio-naïve patients with ulcerative colitis. Due to the exclusion of patients with prior biologic experience in the PURSUIT trials, limited data exists in this population. The aim of our study is to evaluate the efficacy of golimumab in bio-experienced patients (as a second or third line anti-TNF agent) in comparison to bio-naive UC patients.Figure 1Figure 2Figure 3Methods: We performed a retrospective chart review of moderate to severe UC patients treated with golimumab at Fondren IBD clinic in Houston Methodist Hospital. Data for patient demographics, disease duration, location, severity, concomitant immunomodulator therapy and prior biologic use in these patients was collected. Clinical and biochemical parameters used to predict response (UC disease activity index(UCDAI), ESR, CRP, albumin levels) were obtained at first visit prior to initiation of golimumab and at subsequent visits post induction. Categorical and continuous variables were compared using chi square and t-test respectively. All p values calculated were two-tailed with 0.05 alpha level of significance. Results: 20 patients with an average age of 39.4 +/-13.4 years (mean +/- SD) were included in the study. See Table 1. The mean follow-up duration between the 1st and 2nd visit was 127.8 +/-83 days (mean +/- SD). Golimumab was used in 10 bio-experienced patients. Result details are shown in table 2. There was a definite drop in UCDAI scores in our patients (7.75 +/-5.2 to 3.9 +/- 4.6), regardless of them being bio-naïve or experienced. Interestingly, a paradoxical trend of ESR and CRP was noted. Patients receiving golimumab as first line biologic had decreased ESR and increased CRP whereas bio-experienced patients had reduced CRP and higher ESR. Mucosal healing was demonstrated in 3 patients who underwent endoscopy after initiation of golimumab (Mayo endoscopy score of 0). Conclusion: This is the first study which reports the endoscopic efficacy of golimumab in bio-experienced patients. Golimumab seems to be a promising therapy for patients with moderate to severe UC. Our results show that regardless of its use in bio-experienced patients as second or third line anti-TNF therapy, there is a significant improvement in UCDAI scores after treatment. Larger study designs will help better delineate this finding.

Enfermedad inflamatoria intestinal: abordaje conjunto digestivo-dermatológico

Inflammatory bowel disease (IBD) is a complex entity that includes Crohn disease and ulcerative colitis. It is characterized by a chronic proinflammatory state of varying intensity that often leads to considerable morbidity. In the last decade, several therapeutic targets have been identified that are susceptible to the use of biological agents, including anti-tumor necrosis factor alpha antibodies, which are associated with paradoxical psoriasiform reactions in 5% of patients. Decision-making in the management of these cases requires close collaboration between the dermatologist and gastroenterologist. Inflammatory bowel disease is also associated with various other dermatologic and rheumatologic manifestations, and presents a genetic and pathogenic association with psoriasis that justifies both the interdisciplinary approach to these patients and the present review.

Premedication prevents infusion reactions and improves retention rate during infliximab treatment.

Infliximab (IFX) is an anti-tumor necrosis factor-alpha antibody used to treat inflammatory joint diseases. Infusion reactions (IR) can occur during and after intravenous administration and often require discontinuation of IFX therapy. This retrospective study aimed at evaluating the incidence of IR in patients with joint inflammatory diseases receiving IFX with and without premedication. Clinical charts of rheumatoid arthritis, psoriatic arthritis, and ankylosing spondylitis patients receiving IFX from January 2002 to December 2014 were reviewed. Patients receiving only one premedication protocol over time were enrolled and clustered based on the type of premedication as follows: group 1 received no premedication; group 2 received paracetamol, esomeprazole, hydrocortisone, and chlorpheniramine maleate; group 3 received paracetamol, hydoxyzine, ranitidine, and 6-methylprednisolone. Adverse events were recorded during the infusion, in the following hours and at control visits. The charts of 105 patients treated with IFX were selected. IR were observed in 23/51 patients of group 1, in 7/35 patients of group 2, and none of 19 patients in group 3. IR incidence was significantly lower in the second (p=0.021) and third (p<0.001) compared to the first group. The incidence of IR was significantly lower in group 3 than group 2 (p<0.043). Moreover, patients in group 1 had a relative risk of developing an IR 2.5 times higher than group 2. In our experience, the use of premedication significantly reduced the number of IR to IFX. In particular, the combination of paracetamol, hydroxyzine, 6-methylprednisolone and ranitidine was more efficacious than paracetamol, esomeprazole, hydrocortisone, and chlorpheniramine maleate combination protocol.

Early Monitoring of Response (MORE) to Golimumab Therapy Based on Fecal Calprotectin and Trough Serum Levels in Patients With Ulcerative Colitis: A Multicenter Prospective Study.

BackgroundThe treatment of ulcerative colitis (UC) patients with moderate to severe inflammatory activity with anti-tumor necrosis factor alpha (TNFα) antibodies leads to a clinical remission rate of 10% after 8 weeks of therapy. However, it must be taken into account that patient selection in clinical trials clearly influences both response and remission rates. An unsatisfactory response to anti-TNFα medication after week 12 often leads to a discontinuation of treatment. The early prediction of clinical response could therefore help optimize therapy and potentially avoid ineffective treatments.ObjectiveThe aim of this study is to develop an algorithm for optimizing golimumab administration in patients with moderate to severe UC by calculating the probability of clinical response in Week 26 based on data from Week 6.MethodsThe study is designed as a prospective, single-arm, multicenter, non-interventional observational study with no interim analyses and a sample size of 58 evaluable patients. The primary outcome is the prediction of clinical response in Week 26 based on a 50% reduction in fecal calprotectin and a positive golimumab trough level in Week 6.ResultsEnrollment started in October 2014 and was still open at the date of submission. The study is expected to finish in December 2016.ConclusionsThe early identification of patients who are responding to an anti-TNFα antibody is therapeutically beneficial. At the same time, patients who are not responding can be identified earlier. The development of a therapeutic algorithm for identifying patients as responders or non-responders can thus help prescribing physicians to both avoid ineffective treatments and adjust dosages when necessary. This in turn promotes a higher degree of treatment tolerance and patient safety in the case of anti-TNFα antibody administration.ClinicalTrialGerman Clinical Trials Register, Deutsches Register Klinischer Studien DRKS00005940; https://drks-neu.uniklinik-freiburg.de/drks_web/navigate.do?navigationId=trial.HTML&TRIAL_ID=DRKS00005940 (Archived by WebCite at http://www.webcitation.org/6i4Xoo1sH)

Golimumab Therapy in Ulcerative Colitis

Ulcerative colitis is a chronic inflammatory condition of the colon, characterized by diffuse mucosal inflammation and blood-mixed diarrhea. The main treatment has been 5-aminosalicylic acid, steroid, thiopurine, and anti-tumor necrosis factor alpha (TNF-α) antibodies including infliximab, adalimumab, and golimumab. Golimumab, a new anti-TNF-α agent has been recently approved for patients with moderate to severe ulcerative colitis. Its efficacy and safety has been demonstrated in line with infliximab and adalimumab in preclinical and clinical studies. This review will focus on golimumab therapy in ulcerative colitis.

Direct Write Protein Patterns for Multiplexed Cytokine Detection from Live Cells Using Electron Beam Lithography.

Simultaneous detection of multiple biomarkers, such as extracellular signaling molecules, is a critical aspect in disease profiling and diagnostics. Precise positioning of antibodies on surfaces, especially at the micro- and nanoscale, is important for the improvement of assays, biosensors, and diagnostics on the molecular level, and therefore, the pursuit of device miniaturization for parallel, fast, low-volume assays is a continuing challenge. Here, we describe a multiplexed cytokine immunoassay utilizing electron beam lithography and a trehalose glycopolymer as a resist for the direct writing of antibodies on silicon substrates, allowing for micro- and nanoscale precision of protein immobilization. Specifically, anti-interleukin 6 (IL-6) and antitumor necrosis factor alpha (TNFα) antibodies were directly patterned. Retention of the specific binding properties of the patterned antibodies was shown by the capture of secreted cytokines from stimulated RAW 264.7 macrophages. A sandwich immunoassay was employed using gold nanoparticles and enhancement with silver for the detection and visualization of bound cytokines to the patterns by localized surface plasmon resonance detected with dark-field microscopy. Multiplexing with both IL-6 and TNFα on a single chip was also successfully demonstrated with high specificity and in relevant cell culture conditions and at different times after cell stimulation. The direct fabrication of capture antibody patterns for cytokine detection described here could be useful for biosensing applications.

Positioning Therapy for Ulcerative Colitis.

Ulcerative colitis (UC) is a chronic inflammatory condition of the colon, characterized by diffuse mucosal inflammation, bloody diarrhea, and urgency. The mainstay of treatment has been mesalamine agents, steroids, thiopurines, and anti-tumor necrosis factor alpha (TNF-α) antibodies. Over the past several years, new therapies have emerged which have provided clinicians new treatment options as well as new challenges in deciding which treatment is best for their patient at given points in their disease course. These agents include budesonide-Multi-Matrix System (MMX), adalimumab, golimumab, and vedolizumab. In addition, randomized controlled trials have investigated a combination therapy of infliximab and azathioprine and a controlled trial of infliximab versus cyclosporine for intravenous steroid refractory UC. This review will focus on where these agents may be optimally positioned in treatment algorithms for UC.