anti-TNF Antibody Research Articles

Characterization of Myeloid Neoplasms in Inflammatory Bowel Disease

Introduction Patients with inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease (CD) are exposed to chronic systemic inflammation and are at risk for secondary malignancies. While risk is highest for solid tumors or lymphoproliferative disorders, secondary myeloid neoplasms (MN) including acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) are also observed. Clonal hematopoiesis (CH) is associated with systemic inflammatory disease, increased risk of MN, and is enriched in UC (Zhang et al, Exp Hematol 2019). It is not known how the inflammatory milieu of IBD may affect progression to, or severity of, MN. We sought to characterize the disease profiles of patients with IBD diagnosed with MN at our institution in order to identify associations between severity or duration of their inflammatory disease and subtype, molecular, or cytogenetic features of MN. Methods We performed a database query for patients with a diagnosis of AML or MDS or chronic myelogenous leukemia (CML) and a diagnosis of UC or CD seen within the Mount Sinai Health System between 2007 and 2022. Treatment history for IBD, age at diagnosis of IBD and MN, molecular, cytogenetic, and morphologic diagnosis of MN, and pertinent laboratory values were collected for analysis. Results Forty-three patients were found to have a dual diagnosis of IBD and MN. CD was the preceding diagnosis in 70% (30/43) of cases. MDS was diagnosed in 51% (22/43) of all cases. Most patients (71%) were male. Twenty-eight patients had extractable data for further analysis including 12 cases of AML (7 de novo, 3 secondary AML, and 2 acute promyelocytic leukemia), 12 cases of MDS, and 4 cases of CML. The average ages at diagnosis of IBD and MN were 49.0 years (yr) and 61.1 yr, respectively, with an average latency between diagnoses of 14.2 yr. Two patients were diagnosed with IBD after MN. The mean age at diagnosis of AML was younger than at diagnosis of MDS (55.9 vs 68.6 yr, p = 0.05), a difference that was driven by CD patients (52.1 vs 71.1 yr, p = 0.03) but not seen with UC patients (67.4 vs 65.2 yr, p = 0.83). A plot of mutations and clinical characteristics for each analyzed record can be found in Figure 1. The most common therapies for IBD included steroids, 5-ASA, and anti-TNF monoclonal antibodies. There was no association (analysis not shown) of any type or number of IBD therapies with type of MN, disease severity at diagnosis, adverse cytogenetics, or enrichment for specific mutations. Longitudinal complete blood counts were available for 9 patients. At least one cytopenia was found in 89% (8/9) of patients within 3 years of diagnosis of MN. Six patients had anemia (4 CD and 2 UC) with mean hemoglobin 10.5 g/dL (95% CI 9.6 - 11.5), 4 had leukopenia (3 CD and 1 UC), and 4 had thrombocytopenia (2 CD and 2 UC). In 100% (6/6) of patients with anemia there was either normocytosis or macrocytosis, with an average mean corpuscular volume of 93.0 fL (95% CI 89.9 - 96.1) preceding the diagnosis of MN. Conclusion In our cohort, patients with IBD were diagnosed with AML at a younger age than is seen in the general population (55.9 vs 68 yr). Type of IBD, IBD directed therapy, or severity of disease was not associated with type of MN, adverse cytogenetics, or MN risk category. The mutational profile of our cohort mirrors that of the general population of patients diagnosed with MN. A larger longitudinal study is underway to further elucidate the relationship between chronic inflammation in IBD, CH, and risk for progression to MN. Of interest are the types of cytopenias observed in IBD patients as early as three years prior to diagnosis of MN. A normocytic or macrocytic anemia, or thrombocytopenia, was evident prior to diagnosis of MN and is inconsistent with typical IBD-associated thrombocytosis and microcytic anemia. Given established risks of progression to MN, we therefore highlight that patients with IBD who develop atypical cytopenias should be referred for comprehensive hematologic assessment. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Life-threatening bowel complications following anti-tumor necrosis factor antibody therapy for patients with inflammatory bowel disease

Incidence of inflammatory bowel disease (IBD) in Taiwan and the experience of the management of serious complications is much lower than in Western countries. The authors just want to alert readers that anti-tumor necrosis factor (TNF) is not without life-threatening complications. Three serious complications were described, including surgical management and outcomes following the application of anti-TNF monoclonal antibody therapy for IBD. Case 1 involved a 25-year-old man treated with five doses of adalimumab for moderate control of Crohn's disease (CD). Six months later, he experienced severe intestinal obstruction, which necessitated right hemicolectomy with ileocolic anastomosis. He recovered postsurgery. Case 2 involved a 54-year-old man treated with adalimumab for intractable ulcerative colitis. Ten days after the second dose of adalimumab, an emergent subtotal colectomy with ileostomy was performed for a transverse colon perforation with peritonitis. The patient underwent an ileal pouch operation a year later. Case 3 was based on a 16-year-old male patient treated with six doses of an infliximab biosimilar for CD; thereafter, an emergent subtotal colectomy with ileostomy and Hartmann's pouch was performed for peritonitis with the presence of a perforated gastrocolic fistula tract. He had no serious complications following an uneventful recovery period. Creating awareness of serious complications associated with biologic treatments and offering appropriate patient management, including surgical treatment, is beneficial to patients.

Potent anti-inflammatory activity of the lectin-like domain of TNF in joints.

In view of the crucial role of tumor necrosis factor (TNF) in joint destruction, TNF inhibitors, including neutralizing anti-TNF antibodies and soluble TNF receptor constructs, are commonly used therapeutics for the treatment of arthropathies like rheumatoid arthritis (RA). However, not all patients achieve remission; moreover, there is a risk of increased susceptibility to infection with these agents. Spatially distinct from its receptor binding sites, TNF harbors a lectin-like domain, which exerts unique functions that can be mimicked by the 17 residue solnatide peptide. This domain binds to specific oligosaccharides such as N'N'-diacetylchitobiose and directly target the α subunit of the epithelial sodium channel. Solnatide was shown to have anti-inflammatory actions in acute lung injury and glomerulonephritis models. In this study, we evaluated whether the lectin-like domain of TNF can mitigate the development of immune-mediated arthritis in mice. In an antigen-induced arthritis model, solnatide reduced cell influx and release of pro-inflammatory mediators into the joints, associated with reduction in edema and tissue damage, as compared to controls indicating that TNF has anti-inflammatory effects in an acute model of joint inflammation via its lectin-like domain.

Systemic and T cell-associated responses to SARS-CoV-2 immunisation in gut inflammation (STAR SIGN study): effects of biologics on vaccination efficacy of the third dose of mRNA vaccines against SARS-CoV-2.

Immunosuppressed patients with inflammatory bowel disease (IBD) experience increased risk of vaccine-preventable diseases such as COVID-19. To assess humoral and cellular immune responses following SARS-CoV-2 booster vaccination in immunosuppressed IBD patients and healthy controls. In this prospective, multicentre, case-control study, 139 IBD patients treated with biologics and 110 healthy controls were recruited. Serum anti-SARS-CoV-2 spike IgG concentrations were measured 2-16 weeks after receiving a third mRNA vaccine dose. The primary outcome was to determine if humoral immune responses towards booster vaccines differ in IBD patients under anti-TNF versus non-anti-TNF therapy and healthy controls. Secondary outcomes were antibody decline, impact of previous infection and SARS-CoV-2-targeted T cell responses. Anti-TNF-treated IBD patients showed reduced anti-spike IgG concentrations (geometric mean 2357.4 BAU/ml [geometric SD 3.3]) when compared to non-anti-TNF-treated patients (5935.7 BAU/ml [3.9]; p< 0.0001) and healthy controls (5481.7 BAU/ml [2.4]; p< 0.0001), respectively. In multivariable modelling, prior infection (geometric mean ratio 2.00 [95% CI 1.34-2.90]) and vaccination with mRNA-1273 (1.53 [1.01-2.27]) increased antibody concentrations, while anti-TNF treatment (0.39 [0.28-0.54]) and prolonged time between vaccination and antibody measurement (0.72 [0.58-0.90]) decreased anti-SARS-CoV-2 spike antibodies. Antibody decline was comparable in IBD patients independent of anti-TNF treatment and antibody concentrations could not predict breakthrough infections. Cellular and humoral immune responses were uncoupled, and more anti-TNF-treated patients than healthy controls developed inadequate T cell responses (15/73 [20.5%] vs 2/100 [2.0%]; p= 0.00031). Anti-TNF-treated IBD patients have impaired humoral and cellular immunogenicity following SARS-CoV-2 booster vaccination. Fourth dose administration may be beneficial for these patients.

Gastrointestinal toxicity of systemic oncology immunotherapy

Systemic anti-cancer immunotherapy provides a substantial progress in options of current oncology treatment. Yet, this therapeutic approach is potentially associated with a significant gastrointestinal toxicity. The purpose of this paper is to provide a comprehensive review on pathogenesis, clinical features, dia gnostics and therapy of these toxicities. Review of current knowledge: Check-point inhibitors brought a major progress in anti-cancer immunotherapy and improved significantly prognosis of several malignancies (e. g. metastatic malignant melanoma, non-small-cell lung cancer, gastric and colorectal cancers in high-risk population associated with presence of pathogenic mutations, renal cell carcinoma, squamous cell carcinoma of the head and neck and urothelial carcinoma). They include monoclonal antibodies targeting cytotoxic T lymphocyte-associated antigen 4 (CTLA4; e. g. ipilimumab, tremelimumab), programmed death-1 receptor (PD-1; e. g. pembrolizumab, nivolumab) and its ligand PD-L1 (e. gatezolizumab, avelumab). Chimeric antigen receptor (CAR) T-cell therapy is another new option for haematological malignancies and metastatic colorectal cancer. Major symptoms of gastrointestinal toxicity caused by systemic immunotherapy include diarrhoea (20-50%), entero-colitis (1-10%) and laboratory or clinical signs of hepatopathy (~10%). Anti-cancer immunotherapy can be also complicated by infections (Clostridium difficile, Mycoplasma and/ or cytomegalovirus). There is no data on other possible complications so far. However, it can be assumed that these will also include bile acid malabsorption as well as small intestinal bacterial overgrowth syndrome. Treatment of gastrointestinal complications of immunotherapy should be graded according to their severity. It includes symptomatic medications (e. g. loperamide), systemic glucocorticoids and anti-TNF monoclonal antibodies (alone or together with mycofenolate mofetil or tacrolimus in the most severe cases). Awareness of possible complications of systemic anti-cancer immunotherapy is crucial for patients safety. It is mandatory to consider immune-related adverse events, complicating infections, bile acids malabsorption and small intestinal bacterial overgrowth syndrome. Prompt proper dia gnostics and immediate vigorous therapy infl uence the outcome of patients signifi cantly. A strictly individualized approach is indispensable.

S863 Impact of Race on ANTI-TNF Immunogenicity in Patients with Inflammatory Bowel Disease

Introduction: Immunogenicity is a major contributor to anti-tumor necrosis factor (anti-TNF) treatment failure in inflammatory bowel disease (IBD). Immunomodulator and anti-TNF combination therapy is associated with a decreased risk of immunogenicity. Anti-TNF immunogenicity was recently linked to HLA-DQA1*05 genotype. This study aims to determine the impact of race on rates of immunogenicity and treatment outcomes of IBD pts on anti-TNF combination therapy. Methods: This was a single-center, retrospective study of IBD pts who have been treated with immunomodulators and anti-TNF combination therapy between 2012 and 2020. Our primary outcomes were the rates of anti-TNF antibody formation and mean anti-TNF drug levels between Caucasian group(CG) and non-Caucasian group(NCG) on combination therapy. Secondary outcomes included steroid-free clinical remission (SFCR), endoscopic remission (ER) (absence of ulcers/erosions in CD and Mayo endoscopic score ≤ 1 for UC), & normal serum C-reactive protein (CRP) (defined as ≤ 5. mg/L). Continuous variables were analyzed using unpaired student’s t-test. Categorical variables were analyzed using a chi-square test. Results: A total of 124 pts were included (CD; 68.5%, UC; 27.4%, indeterminate colitis; 3.2%, pouchitis; 0.9%). The median age was 32 years (range 13-69), and 54.8% were male. A total of 87 pts were on infliximab & 37 pts were on adalimumab. Combination therapy with thiopurine was employed in 87.1% while 12.9% were on methotrexate. A total of 85 pts were self-identified as Caucasian. There were no significant differences between CG vs NCG in terms of baseline and disease characteristics (Table). Anti-TNF antibody formation was observed in 32.9% of pts in the CG, and 20.5% of pts in the NCG (p=0.16). Mean anti-TNF drug levels were lower in the CG at 15.4 ± 14.5 µg/mL vs. the NCG at 22.7 ± 15.1 µg/mL (p=0.01). SFCR was observed in 61.2% vs. 58.9% (p=0.82) in CG vs. NCG, respectively. In CG 42.4% vs. 33.3% in NCG (p=0.34) discontinued anti-TNF treatment during follow-up. ER was observed in 45.9 % vs 51.3% (p=0.44), and normal CRP was observed in 51.8% vs. 58.9% (p=0.28) in CG vs. NCG, respectively. Conclusion: In our cohort, Caucasian pts on anti-TNF combination therapy for IBD had significantly lower anti-TNF drug levels as compared to the non-Caucasian group. However, there was no significant difference between rates of anti-TNF antibody formation and clinical outcomes between the groups. Larger studies are needed to clarify impact of race on anti-TNF therapy. Table 1. - Comparison of Characteristics Between Caucasian vs. Non-Caucasian Race Caucasian (n=85), n (%), mean (SD) Non-Caucasian (n=39), n (%), mean (SD) P-value 34.7 (13.7) 34.7 (11.7) 0.98 47 (55.3) 21 (53.9) 0.88 7 (8.2) 6 (15.4) 0.23 36.4 (27.1) 41.9 (31.8) 0.35 0.07 53 (62.4) 32 (82.1) 29 (34.1) 5 (12.8) 2 (2.3) 2 (5.1) 1 (1.2) 0 21 (24.7) 15 (38.5) 0.12 24 (28.2) 14 (35.9) 0.81 10 (11.8) 6 (15.4) 0.77 61 (71.8) 26 (66.7) 0.56 69 (81.2) 29 (74.4) 0.39 12.5 (21.4) 6.8 (7.6) 0.15 3.9 (0.6) 4.1 (0.4) 0.16 *Data missing for 68 patients.**Data missing for 42 patients.Abbreviations: Anti TNF, anti-tumor necrosis factor; CRP, C reactive protein.

S970 Older Adults Are at Higher Risk for Developing Anti-TNF Antibodies

S970 Older Adults Are at Higher Risk for Developing Anti-TNF Antibodies

33431 Efficacy of 80 mg/week dose of adalimumab (Humira) in treatment of hidradenitis suppurativa

Hidradenitis suppurativa (HS) is an inflammatory skin condition involving the apocrine gland-bearing regions, such as the axillae, groin, anogenital, and inframammary areas. Though its etiology is unknown, disease manifests when hair follicles become obstructed, leading to the release of inflammatory mediators, such as TNF-α and IL-17. Levels of TNF-α have been found to be elevated in lesions of HS, providing a basis for the use of biologics in treatment of severe HS. Adalimumab is a recombinant human IgG1 anti-TNF monoclonal antibody which acts via antagonism of TNF-α.

33431 Efficacy of 80 mg/week dose of adalimumab (Humira) in treatment of hidradenitis suppurativa

Hidradenitis suppurativa (HS) is an inflammatory skin condition involving the apocrine gland-bearing regions, such as the axillae, groin, anogenital, and inframammary areas. Though its etiology is unknown, disease manifests when hair follicles become obstructed, leading to the release of inflammatory mediators, such as TNF-α and IL-17. Levels of TNF-α have been found to be elevated in lesions of HS, providing a basis for the use of biologics in treatment of severe HS. Adalimumab is a recombinant human IgG1 anti-TNF monoclonal antibody which acts via antagonism of TNF-α. Although adalimumab has been studied in phase 2 and 3 trials and has been approved as a treatment modality for HS at a 40 mg weekly dose, numerous patients still suffer on a day-to-day basis due to the severity of their disease and do not respond to this conventional dosing. We present a series of 6 such patients whose response to an 80 mg/week dose of adalimumab has been positive, with a decrease in severity and number of HS lesions, leading to improvement in the Hidradenitis Suppurativa Clinical Response (HiSCR) in the absence of serious adverse effects. Although this dosing regimen has not been well-documented in literature, it is important to consider the lasting benefits of treatment with a higher dose in individuals afflicted with treatment-resistant HS, including a decreased likelihood of comorbidities and complications and an improved quality of life.

Implications for sequencing of biologic therapy and choice of second anti-TNF in patients with inflammatory bowel disease: results from the IMmunogenicity to Second Anti-TNF therapy (IMSAT) therapeutic drug monitoring study.

Anti-drug antibodies are associated with treatment failure to anti-TNF agents in patients with inflammatory bowel disease (IBD). To assess whether immunogenicity to a patient's first anti-TNF agent would be associated with immunogenicity to the second, irrespective of drug sequence METHODS: We conducted a UK-wide, multicentre, retrospective cohort study to report rates of immunogenicity and treatment failure of second anti-TNF therapies in 1058 patients with IBD who underwent therapeutic drug monitoring for both infliximab and adalimumab. The primary outcome was immunogenicity to the second anti-TNF agent, defined at any timepoint as an anti-TNF antibody concentration ≥9AU/ml for infliximab and ≥6AU/ml for adalimumab. In patients treated with infliximab and then adalimumab, those who developed antibodies to infliximab were more likely to develop antibodies to adalimumab, than patients who did not develop antibodies to infliximab (OR 1.99, 95%CI 1.27-3.20, p=0.002). Similarly, in patients treated with adalimumab and then infliximab, immunogenicity to adalimumab was associated with subsequent immunogenicity to infliximab (OR 2.63, 95%CI 1.46-4.80, p < 0.001). For each 10-fold increase in anti-infliximab and anti-adalimumab antibody concentration, the odds of subsequently developing antibodies to adalimumab and infliximab increased by 1.73 (95% CI 1.38-2.17, p < 0.001) and 1.99 (95%CI 1.34-2.99, p < 0.001), respectively. Patients who developed immunogenicity with undetectable drug levels to infliximab were more likely to develop immunogenicity with undetectable drug levels to adalimumab (OR 2.37, 95% CI 1.39-4.19, p < 0.001). Commencing an immunomodulator at the time of switching to the second anti-TNF was associated with improved drug persistence in patients with immunogenic, but not pharmacodynamic failure. Irrespective of drug sequence, immunogenicity to the first anti-TNF agent was associated with immunogenicity to the second, which was mitigated by the introduction of an immunomodulator in patients with immunogenic, but not pharmacodynamic treatment failure.

Behçet's Disease-related Budd-Chiari Syndrome Successfully Managed with anti-TNF Antibody: A Case Report and Review of the Literature.

Behçet’s disease (BD) is multisystemic vasculitis with heterogeneous clinical manifestations. We describe the case of a 26-year-old man who presented with Budd–Chiari syndrome (BCS) related to BD. The patient received infliximab (IFX) due to the severity of vascular involvement. Subsequently, after IFX therapy, hospital-acquired pneumonia, trapped lung, and fungal infection of the lung and central nervous system developed as complications. The patient benefited from a second course of IFX and clinical remission was achieved following early identification and treatment of complications. Data on the presentation and prognosis of BCS related to BD are extremely limited. Our case report supports the growing evidence that anti-TNF antibody is a promising treatment for BD-related BCS.LEARNING POINTSBehçet’s disease-related Budd–Chiari syndrome is a rare form of vascular involvement that severely affects mortality.Behçet’s disease-related Budd–Chiari syndrome is frequently confused with idiopathic thrombosis and may be underdiagnosed.Infliximab could be a therapeutic option for refractory Behçet’s disease with major vascular involvement, but the increased risk of opportunistic infections should be kept in mind.

An Overview of Tumor Necrosis Factor-α on Pathophysiological Mechanisms, Relevant Therapeutic Status in Breast Cancer

TNFα is an essential pro-inflammatory cytokine that is prevalent in the tumor microenvironment and is involved in mediating or activating many significant signaling pathways which result in inflammation, apoptosis, and tumor cell proliferation, survival, and invasiveness. In breast cancer, TNFα is involved throughout all stages from occurrence, development, procession, and metastasis to recurrence. Researchers have pointed out that TNFα plays a major role in the estrogen biosynthesis pathway, especially in the process of adipose tissue switching to estrogen. In the breast tumor microenvironment, TNFα may participate in the mediation of estrone sulfatase expression and activity. In terms of therapeutics, methods to suppress TNFα signaling in breast cancer have been proposed. To neutralize the pro-tumor and inflammatory effects of TNFα, most research opts to use anti-TNFα antibodies. According to the research, the administration of TNFα antagonists can suppress the development of breast cancer cells and strengthen the chemotherapeutic response when used as adjuvant therapy with chemotherapy. Consequently, tumor drug resistance can be well controlled. However, some side effects like systemic toxicity, the typical skin lesion, and the increasing risk of developing new cancers are still major issues. More extensive clinical trials have to be carried out for deeper investigation. This paper gives an overview of the intrinsic features of TNFα as a cytokine and gets insight into the pathophysiological mechanisms mediated by TNFα in breast cancer. Furthermore, the current state of knowledge in terms of TNF-related therapeutic strategies was adequately summarized and discussed.

Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display.

Antibody stability and affinity are two important features of its applications in therapy and diagnosis. Antibody display technologies such as yeast and bacterial displays have been successfully used for improving both affinity and stability. Although mammalian cell display has also been utilized for maturing antibody affinity, it has not been applied for improving antibody stability. Previously, we developed a Chinese hamster ovary (CHO) cell display platform in which activation-induced cytidine deaminase (AID) was used to induce antibody mutation, and antibody affinity was successfully matured using the platform. In the current study, we developed thermo-resistant (TR) CHO cells for the purpose of maturing both antibody stability and affinity. We cultured TR CHO cells displaying an antibody mutant library and labeled them at temperatures above 41 °C, enriching cells that displayed antibody mutants with both the highest affinities and the highest display levels. To evaluate our system, we chose three antibodies to improve their affinities and stabilities. We succeeded in simultaneously improving both affinities and stabilities of all three antibodies. Of note, we obtained an anti-TNFα antibody mutant with a Tm (dissolution temperature) value 12 °C higher and affinity 160-fold greater than the parent antibody after two rounds of cell proliferation and flow cytometric sorting. By using CHO cells with its advantages in protein folding, post-translational modifications, and code usage, this procedure is likely to be widely used in maturing antibodies and other proteins in the future.

Effects of Anti-Tumor Necrosis Factor Therapy on Osteoblastic Activity at Sites of Inflammatory and Structural Lesions in Radiographic Axial Spondyloarthritis: A Prospective Proof-of-Concept Study Using Positron Emission Tomography/Magnetic Resonance Imaging of the Sacroiliac Joints and Spine.

Proof-of-concept trial to determine the effects of tumor necrosis factor inhibitor (TNFi) therapy on osteoblastic activity at sites of inflammatory and structural lesions in patients with radiographic axial spondyloarthritis (SpA), using fluorine 18-labeled NaF (18 F-NaF) positron emission tomography/magnetic resonance imaging (PET/MRI). Sixteen patients with clinically active radiographic axial SpA were prospectively enrolled to receive TNFi treatment and undergo 18 F-NaF PET/MRI of the sacroiliac (SI) joints and spine at baseline and at a follow-up visit 3-6 months after treatment initiation. Three readers (1 for PET/MRI and 2 for conventional MRI) evaluated all images, blinded to time point. Bone marrow edema, structural lesions (i.e., fat lesions, sclerosis, erosions, and ankylosis), and 18 F-NaF uptake at SI joint quadrants and vertebral corners (VCs) were recorded. Overall, 11 male and 5 female patients (mean age ± SD 38.6 ± 12.0 years) were followed up for a mean duration of 4.6 months (range 3-6). 18 F-NaF PET/MRI was conducted on SI joints for 16 patients and the spine for 10; 128 SI joint quadrants and 920 VCs were analyzed at each time point. At baseline, 18 F-NaF uptake was demonstrated in 96.0% of SI joint quadrants with bone marrow edema, 94.2% with sclerosis, and 88.3% with fat lesions. At follow-up, 65.3% of SI joint quadrants with bone marrow edema (P < 0.001), 33.8% with sclerosis (P = 0.23), and 24.5% with fat lesions (P = 0.01) had less 18 F-NaF uptake, compared with baseline. For VCs, 18 F-NaF uptake at baseline was found in 81.5% of edges with sclerosis, 41.9% with fat lesions, and 33.7% with bone marrow edema. At follow-up, 73.5% of VCs with bone marrow edema (P = 0.01), 53.3% with fat lesions (P = 0.03), and 55.6% with sclerosis (P = 0.16) showed less 18 F-NaF uptake, compared with baseline. Anti-TNF antibody treatment led to a significant decrease in osteoblastic activity within 3-6 months, especially, but not solely, at sites of inflammation. Larger data sets are needed for confirmation of the antiosteoblastic effects of TNFi for the prevention of radiographic progression in axial SpA.

Persistent popliteal lymphatic muscle cell coverage defects despite amelioration of arthritis and recovery of popliteal lymphatic vessel function in TNF-Tg mice following anti-TNF therapy

While rheumatoid arthritis patients and tumor necrosis factor transgenic (TNF-Tg) mice with inflammatory-erosive arthritis display lymphatic drainage deficits, the mechanisms responsible remain unknown. As ultrastructural studies of joint-draining popliteal lymphatic vessels (PLVs) in TNF-Tg mice revealed evidence of lymphatic muscle cell (LMC) damage, we aimed to evaluate PLV-LMC coverage in TNF-Tg mice. We tested the hypothesis that alpha smooth muscle actin (αSMA)+ PLV-LMC coverage decreases with severe inflammatory-erosive arthritis, and is recovered by anti-TNF therapy facilitated by increased PLV-LMC turnover during amelioration of joint disease. TNF-Tg mice with established disease received anti-TNF monoclonal antibody (mAb) or placebo IgG isotype control mAb therapy (n = 5) for 6-weeks, while wild-type (WT) littermates (n = 8) received vehicle (PBS). Bromodeoxyuridine (BrdU) was also administered daily during the treatment period to monitor PLV-LMC turnover. Effective anti-TNF therapy was confirmed by longitudinal assessment of popliteal lymph node (PLN) volume via ultrasound, PLV contraction frequency via near-infrared imaging of indocyanine green, and ankle bone volumes via micro-computed tomography (micro-CT). Terminal knee micro-CT, and ankle and knee histology were also performed. PLVs were immunostained for αSMA and BrdU to evaluate PLV-LMC coverage and turnover, respectively, via whole-mount fluorescent microscopy. Anti-TNF therapy reduced PLN volume, increased talus and patella bone volumes, and reduced tarsal and knee synovial areas compared to placebo treated TNF-Tg mice (p < 0.05), as expected. Anti-TNF therapy also increased PLV contraction frequency at 3-weeks (from 0.81 ± 1.0 to 3.2 ± 2.0 contractions per minute, p < 0.05). However, both anti-TNF and placebo treated TNF-Tg mice exhibited significantly reduced αSMA+ PLV-LMC coverage compared to WT (p < 0.05). There was no correlation of αSMA+ PLV-LMC coverage restoration with amelioration of inflammatory-erosive arthritis. Similarly, there was no difference in PLV-LMC turnover measured by BrdU labeling between WT, TNF-Tg placebo, and TNF-Tg anti-TNF groups with an average of < 1% BrdU+ PLV-LMCs incorporated per week. Taken together these results demonstrate that PLV-LMC turnover in adult mice is limited, and that recovery of PLV function during amelioration of inflammatory-erosive arthritis occurs without restoration of αSMA+ LMC coverage. Future studies are warranted to investigate the direct and indirect effects of chronic TNF exposure, and the role of proximal inflammatory cells on PLV contractility.

618 Macrophage depletion preserves dermal collagen in UVB exposed mice

We previously reported that anti-TNFα antibodies (etanercept) block UVB-induced recruitment of neutrophils and macrophages (MΦ) into the dermis in mice. Paradoxically, etanercept accelerates loss of dermal collagen. To examine the role of MΦ in dermal collagen alteration during UVB exposure, we depleted MΦ in mice. C57BL/6J mice were treated with (0.15 ml) of Clophosome-A-Clodronate Liposomes (CCL) i.p. twice, 5 and 3 days before UVB exposure. We had 4 groups- sham, sham+CCL, UVB- and UVB+CCL-treated mice.

Production of an anti-TNFα antibody in murine myeloma cells by perfusion culture.

Infliximab is a mouse/human chimeric IgG1 monoclonal antibody which recognizes the proinflammatory cytokine, tumor necrosis factor α (TNFα), and inhibits receptor interactions, thereby decreasing inflammation and autoimmune response in patients. This monoclonal antibody has been successfully used to treat rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis. However, the high treatment cost limits patient access to this biotherapy. One alternative to this problem is the use of biosimilars. In this work, we describe the stable expression and physicochemical characterization of an anti-TNFα antibody. While infliximab is produced in recombinant murine SP2/0 cells, our anti-TNFα IgG antibody was expressed in recombinant murine NS0 myeloma cells. The best anti-TNFα antibody-expressing clone was selected from three clone candidates based on the stability of IgG expression levels, specific productivity as well as TNFα-binding activity compared to commercial infliximab. Our results indicate that the selected cell clone, culture medium, and fermentation mode allowed for the production of an anti-TNFα antibody with similar characteristics to the reference commercially available product. An optimization of the selected culture medium by metabolomics may increase the volumetric productivity of the process to satisfy the demand for this product. Further experiments should be performed to evaluate the biological properties of this anti-TNFα antibody. KEY POINTS: • An anti-TNFα antibody was produced in NS0 cells using perfusion culture. • A proprietary chemically defined culture medium was used to replace commercially available protein-free medium. • The purified anti-TNFα antibody was comparable to the reference marketed product.

Disease-specific expansion of CD29+IL-17RA+ T effector cells possessing multiple signalling pathways in spondyloarthritis.

T cells adhere to enthesis fibrocartilage via integrins and intrinsically require IL-17RA-mediated signals to maintain their effector function. We analysed CD29+IL-17RA+ T cells in inflamed lesions and peripheral blood in patients with SpA and investigated their association with disease activity and therapeutic response. Transcriptome analysis of synovial fluid T cells from PsA was performed using publicly available bulk cell RNA sequencing data. Blood samples were obtained from healthy controls (n = 37), RA (n = 12), IgG4-related disease (IgG4-RD; n = 12), large vessel vasculitis (LVV; n = 12) and SpA (n = 28) and were analysed by flow cytometry. T cells in the inflamed joints of PsA showed CD29 and IL-17RA expression. CD29+IL-17RA+ T cells showed enriched CXCR3+CD45RA+ effector cells and activation of spleen tyrosine kinase (Syk), nuclear factor κB (NF-κB) and Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathways. The proportion of peripheral blood CD29+IL-17RA+ T cells was significantly increased in patients with SpA compared with patients with RA, IgG4-RD or LVV and in healthy controls. Based on the ASDAS-CRP scores, the proportion of CD29+IL-17RA+ T cells was positively correlated with disease activity in treatment-naïve patients with active SpA. Anti-IL-17 but not anti-TNF monoclonal antibodies reduced CD29+IL-17RA+ T cells. CD29+IL-17RA+ T effector cells with enhanced Syk, NF-κB and JAK-STAT pathways were specifically increased in SpA and were correlated with disease activity, implicating a role of this newly identified T cell population in the pathogenesis. Anti-IL-17 monoclonal antibodies may be effective for patients by reducing this pathogenic T cell population.

A Randomized Pharmacokinetic Study in Healthy Male Subjects Comparing a High-concentration, Citrate-free SB5 Formulation (40mg/0.4ml) and Prior SB5 (Adalimumab Biosimilar).

IntroductionSB5 is an approved biosimilar of adalimumab, a monoclonal anti-tumor necrosis factor (anti-TNF) antibody. This study compared pharmacokinetics (PK), safety, tolerability, and immunogenicity between a new high-concentration, low-volume, and citrate-free formulation (40 mg/0.4 ml, SB5-HC) and the current low-concentration formulation with higher volume (40 mg/0.8 ml, SB5-LC) to evaluate the bioequivalence of the two formulations.MethodsThis study was a randomized, single-blind, two-arm, parallel-group, single-dose study in healthy male subjects. Subjects were randomized to receive either SB5-HC or SB5-LC via subcutaneous injection using a pre-filled syringe. Primary endpoints were the area under the curve of the concentration–time curve from zero to infinity (AUCinf) and maximum serum concentration (Cmax). Bioequivalence was achieved if the 90% confidence intervals (CIs) for the ratios of the geometric least squares mean (LSMean) of primary endpoints were within the pre-defined bioequivalence margins of 0.80–1.25. Secondary endpoints included safety, tolerability, and immunogenicity.ResultsSubjects (n = 188) were randomized to SB5-HC (n = 94) or SB5-LC (n = 94). Baseline characteristics were comparable between the two treatment groups. The mean values for AUCinf and Cmax were similar between the SB5-HC and SB5-LC groups. For the primary endpoints, the geometric LSMean ratios (90% CI) for AUCinf and Cmax were 0.920 (0.8262–1.0239) and 0.984 (0.9126–1.0604), respectively, placing the corresponding 90% CIs well within the pre-defined bioequivalence margin of 0.80–1.25. All treatment-emergent adverse events (TEAEs) were considered mild to moderate and were reported for 44.7% and 51.1% of subjects in the SB5-HC and SB5-LC groups, respectively. Immunogenicity assessed by frequency of occurrence of anti-drug antibodies (ADAs) and neutralizing antibodies (NAbs) was comparable between groups.ConclusionsThis bridging study demonstrated PK equivalence and comparable safety and tolerability of subcutaneous injection of SB5 via SB5-HC or SB5-LC.Clinicaltrials.gov identifierhttps://clinicaltrials.gov/ct2/show/NCT04514796.Supplementary InformationThe online version contains supplementary material available at 10.1007/s40744-022-00471-8.

Development of a Machine Learning Model to Predict Non-Durable Response to Anti-TNF Therapy in Crohn's Disease Using Transcriptome Imputed from Genotypes.

Almost half of patients show no primary or secondary response to monoclonal anti-tumor necrosis factor α (anti-TNF) antibody treatment for inflammatory bowel disease (IBD). Thus, the exact mechanisms of a non-durable response (NDR) remain inadequately defined. We used our genome-wide genotype data to impute expression values as features in training machine learning models to predict a NDR. Blood samples from various IBD cohorts were used for genotyping with the Korea Biobank Array. A total of 234 patients with Crohn’s disease (CD) who received their first anti-TNF therapy were enrolled. The expression profiles of 6294 genes in whole-blood tissue imputed from the genotype data were combined with clinical parameters to train a logistic model to predict the NDR. The top two and three most significant features were genetic features (DPY19L3, GSTT1, and NUCB1), not clinical features. The logistic regression of the NDR vs. DR status in our cohort by the imputed expression levels showed that the β coefficients were positive for DPY19L3 and GSTT1, and negative for NUCB1, concordant with the known eQTL information. Machine learning models using imputed gene expression features effectively predicted NDR to anti-TNF agents in patients with CD.