Search

ObjectivesThis study aimed to clarify the usefulness of screening for malignancies using CT before the initiation of biologic and targeted synthetic DMARDs (b/tsDMARDs) in patients with active RA.MethodsWe examined 2192 patients with RA who underwent plain CT scans prior to the initiation of b/tsDMARDs. The sensitivity for detecting malignancy was measured and compared with that of regular screening (physical examination and X-ray). We then evaluated the clinical characteristics, prognosis and treatment of patients with RA with concomitant malignancies. Additionally, we determined the incidence rate of malignancy in patients with RA who were initiated on b/tsDMARDs after CT screening.ResultsOf the 2192 patients, 33 (1.5%) were diagnosed with malignancy after CT screening. Whereas regular screening detected only seven malignancies, CT screening further detected 26 (including 19 at the early stage). On the other hand, 86% of the malignancies detectable by regular screening were at an advanced stage. Patients diagnosed with early-stage malignancies received RA treatments that included b/tsDMARDs after curative resection; 80% of these patients achieved low disease activity after 1 year. This rate was comparable to the patients without malignancy detection after screening (70%). The 5 year incidence of malignancy after the initiation of b/tsDMARDs after CT screening was lower than that of the RA cohort without CT screening (standardized incidence ratio: 0.35).ConclusionScreening in patients with RA using CT before the initiation of b/tsDMARDs allows for the early detection and treatment of malignancy, resulting in safer and more stable b/tsDMARD treatments.

ObjectiveThis study aimed to understand the role of mammalian target of rapamycin (mTOR) in CD8+ cells in the pathogenicity of RA and the changes after treatment with biologic drugs.MethodsPeripheral blood mononuclear cells (PBMCs) were isolated from 17 healthy controls and 86 patients with RA. Phosphorylation of mTOR (p-mTOR) and its clinical relevance were evaluated. The role of mTOR in CD8+ cells was also examined in vitro.ResultsPatients with RA who had a moderate or high disease activity, were biologic-naïve, and were refractory to MTX were enrolled in this study. The p-mTOR levels in CD8+ cells were higher in patients with RA than in healthy controls, and they positively correlated with the disease activity in such patients. However, after one year of treatment with TNF inhibitors, the p-mTOR levels in CD8+ cells were suppressed and showed a positive correlation with the treatment response, which was not observed in the abatacept-treatment group. In vitro stimulation of CD8+ cells with anti-CD3 and anti-CD28 antibodies induced mTOR phosphorylation and increased the production of granzyme B, granulysin, TNF-α and IFN-γ but decreased the production of granzyme K. However, on treatment with TNF inhibitors, p-mTOR levels in CD8+ cells and granzyme B production decreased, while granzyme K production increased. The production of granulysin and IFN-γ was not affected by the TNF inhibitors.ConclusionThese results suggested that mTOR activation in CD8+ cells may be a novel evaluation marker for RA disease activity and a predictive marker of therapeutic response to TNF inhibitors.

Objective To evaluate the dysfunction of B-cell metabolism and its involvement in SLE pathology. Methods We assessed the expression of metabolic markers of B cells in the peripheral blood of healthy controls (HCs) and SLE patients by using flow cytometry. In vitro, peripheral B cells were isolated from HCs and SLE patients to investigate the metabolic regulation mechanisms involved in their differentiation. Results The expression level of DiOc6 (mitochondrial membrane hyperpolarization) was higher in B cells from SLE patients than in HCs, and correlated to the percentage of plasmablasts in CD19+ cells and with SLEDAI, a disease activity score. Stimulation of CD19+ cells with the Toll-like receptor 9 (TLR9) ligand CpG and IFN-α enhanced glycolysis, oxidative phosphorylation (OXPHOS), DiOc6 expression, and plasmablast differentiation in vitro. In the absence of glutamine, both glycolysis and OXPHOS were reduced, and plasmablast differentiation was suppressed, whereas there was no change in the absence of glucose. As glutamine is an important nutrient for protein synthesis, we further investigated the effect of the glutaminase inhibitor BPTES, which inhibits glutamine degradation, on metabolic regulation. BPTES reduced DiOc6 expression, OXPHOS, reactive oxygen species (ROS) production, adenosine triphosphate (ATP) production, plasmablast differentiation without affecting glycolysis. Metformin inhibited CpG- and IFN-α-induced glutamine uptake, mitochondrial functions and suppressed plasmablast differentiation. Conclusions Mitochondrial dysfunction in B cells is associated with plasmablast differentiation and disease activity in SLE. Enhanced mitochondrial functions mediated by glutamine metabolism are important for plasmablast differentiation, which may be a potential therapeutic target for SLE.

Objective Plasmablasts play important roles in autoimmune diseases, including systemic lupus erythematosus (SLE). Activation of mechanistic target of rapamycin complex 1 (mTORC1) is regulated by amino acid levels. In patients with SLE, mTORC1 is activated in B cells and modulates plasmablast differentiation. However, the detailed mechanisms of amino acid metabolism in plasmablast differentiation remain elusive. We undertook this study to evaluate the effects of methionine in human B cells. Methods Purified CD19+ cells from healthy donors (n = 21) or patients with SLE (n = 35) were cultured with Toll-like receptor 7/9 ligand, interferon-α (IFNα), and B cell receptor crosslinking, and we determined the types of amino acids that were important for plasmablast differentiation and amino acid metabolism. We also identified the transcriptional regulatory mechanisms induced by amino acid metabolism, and we assessed B cell metabolism and its relevance to SLE. Results The essential amino acid methionine strongly committed cells to plasmablast differentiation. In the presence of methionine, Syk and mTORC1 activation synergistically induced methyltransferase EZH2 expression. EZH2 induced H3K27me3 at BTB and CNC homolog 2 (Bach2) loci and suppressed Bach2 expression, leading to induction of B lymphocyte-induced maturation protein 1 and X-box binding protein 1 expression and plasmablast differentiation. CD19+ cells from patients with SLE overexpressed EZH2, which was correlated with disease activity and autoantibody production. Conclusion Our findings show that methionine activated signaling by controlling immunologic metabolism in B cells and played an important role in the differentiation of B cells into plasmablasts through epigenome modification of Bach2 by the methyltransferase EZH2.

Microscopic polyangiitis (MPA) is a type of necrotizing vasculitis associated with high levels of myeloperoxidase-specific antineutrophil cytoplasmic antibody (MPO-ANCA). While generally associated with renal dysfunction, MPA can also cause intraabdominal hemorrhage in rare cases. A 66-year-old man was admitted to our hospital for renal dysfunction, numbness, and weight loss for 3 months. He had no significant medical history. Renal biopsy revealed crescentic glomerulonephritis with necrotizing vasculitis, which was associated with a high serum titer of MPO-ANCA, leading to a diagnosis of MPA. Remission-induction treatment with glucocorticoids and rituximab was initiated, which improved the patient’s general condition and renal failure. His blood pressure was elevated and was controlled by amlodipine treatment. Two months after discharge, he visited the emergency department because of chest pain. A diagnosis of acute cardiovascular syndrome was suggested; however, his cardiac artery was not stenotic. The patient’s blood pressure was high despite antihypertensive therapy, and he developed posterior reversible encephalopathy syndrome (PRES). Despite intensive treatment, the patient died 3 days later. An autopsy revealed that the cause of death was hypovolemic shock due to massive intra-abdominal hemorrhage from the ruptured mesenteric artery involved in vasculitis. In cases of MPA with sudden-onset chest or abdominal pain, a ruptured intra-abdominal artery should be considered. Secondary hypertension associated with vasculitis should be carefully managed to prevent hemorrhagic complications and PRES.

ObjectivesB cells play an important pathological role in RA. In this study, we investigated the role of metabolic regulator mTOR in B cells and its relevance to the pathology of RA.MethodsPeripheral blood mononuclear cells were isolated from 31 normal subjects and 86 RA patients and the gated B cells were assessed for mTOR phosphorylation and chemokine receptor expression. In vitro studies on peripheral blood B cells isolated from the control and RA patients investigated the molecular mechanisms.ResultsHigher concentrations of CXCL10 (CXCR3 ligands) and lower percentages of CXCR3+ memory B cells were present in the peripheral blood of RA patients relative to the control. RA patients with high CXCL10 concentrations had smaller percentage of CXCR3+ memory B cells and high disease activity. One-year treatment with TNF inhibitors increased the percentage of CXCR3+ memory B cells and reduced serum CXCL10 concentrations. mTOR phosphorylation in B cells was further enhanced in RA patients, compared with the control, and was selectively enhanced in CXCR3+ memory B cells. mTOR phosphorylation in CXCR3+ memory B cells correlated with disease activity. In vitro, mTOR phosphorylation in B cells enhanced IL-6 production and increased RANKL expression.ConclusionmTOR activation in CXCR3+ memory B cells of RA patients is associated with disease activity, mediated through IL-6 production and RANKL expression. The obtained results also suggest that TNF inhibitors mediate an impact on the association between CXCL10 and mTOR activated CXCR3+ memory B cells.

We herein report a 65-year-old man with elevated serum IgG4 levels, enlarged thyroid, and renal dysfunction, mimicking IgG4-related disease (IgG4-RD). The definitive diagnosis of IgG4-RD was not established because a tissue biopsy revealed no IgG4-positive cell infiltration or fibrosis. The presence of an M peak in the β fraction, Bence Jones protein in urine, and progressive anemia suggested multiple myeloma (MM). The κ/λ ratio was >100, tumor plasma cells were present at >20% in bone marrow, and immunostaining revealed IgG4-positive plasma cells; therefore, he was diagnosed with IgG4-type MM. Patients with elevated IgG4 levels with no significant mass lesions should undergo systemic examinations to exclude malignancy.

M. abscessus is a rapidly growing mycobacteria (RGM) and is the most common cause of pulmonary RGM infection. M. abscessus pleurisy is extremely rare. We herein report the case of a young patient with M. abscessus pleurisy without any lung lesions. A laboratory analysis of the pleural effusion revealed lymphocyte predominance and increased adenosine deaminase, similar to the findings observed in tuberculous pleurisy. The patient was initially treated for tuberculous pleurisy, which resulted in the partial improvement of the patient's symptoms and pleural effusion. M. abscessus pleurisy should be considered, especially in immunocompromised individuals, even in the absence of pulmonary involvement.

Background: B cells play an important role in systemic lupus erythematosus (SLE). Several research protocols have focused in recent years on the topic of immunometabolism. Activation of immunocompetent cells depends on rapid synthesis of cell structure components and biomolecules, which requires enormous amounts of energy, nucleic acids and lipids. Amino acids are important ingredients of many metabolic processes. However, the role of amino acids in plasmablast differentiation and their relevance to the pathogenesis of SLE remain elusive. Objectives: To determine the role of essential amino acids in human B cell differentiation and relevance to the pathogenesis of SLE. Methods: In the in vitro arm of the study, purified CD19+ B cells from healthy donors were cultured with TLR7/9 ligand (LOX or CpG), IFN-α and B cell receptor (BCR) cross-linking, in the presence or absence of amino acids. We determined 1) the types of amino acids that are important for PB differentiation, 2) the amino acid transporters that are important for PB differentiation, 3) the main signaling pathway(s) involved in the presence of amino acids, 4) the transcriptional factors used in the presence of amino acids. In the clinical arm of the study, peripheral blood mononuclear cells (PBMCs) were obtained from 24 patients with RA, 35 patients with SLE, and 28 age-matched healthy controls, and subjected to flow cytometric analysis to determine the expression of amino acids-related markers. Results: 1) Stimulation with the combination of BCR, IFN-α and TLR7/9 ligand induced PB differentiation accompanied by uptake of amino acids. PB differentiation was abrogated in the absence of essential amino acid methionine, and to a lesser extent leucine, but not in non-essential amino acid cystine. 2) LAT1 and CD98 are known amino acid transporters. Stimulation with BCR, IFN-α and TLR7/9 ligand induced CD98 expression but suppressed LAT1 expression. CD98 expression was higher in CD27hiCD38hi PB than in CD27-CD38- non-PB. 3) Previous studies reported that amino acids were perceived by the sensor, leading to mTORC1 phosphorylation. However, the mechanism by which amino acids activate other intracellular signaling pathways in B cells remains elusive. We found that methionine facilitated both the BCR and mTORC1 signals. In addition, the two signals synergistically induced EZH2 expression, which is well known as a transcriptional factor for histone modification via induction of H3K27me3, in the presence of methionine. 4) Methionine induced EZH2 expression, leading to suppression of BACH2, induction of BLIMP1, XBP1 and PB differentiation. These results indicate that EZH2 is a critical factor for PB differentiation in the presence of methionine. Assessment of the expression of amino acid transporters CD98, LAT1 and EZH2 in B cells in RA and SLE patients showed overexpression of CD98 and EZH2, but not LAT1, in SLE, compared with RA and control. In SLE patients, EZH2 expression level correlated with that of CD98 in B cells. EZH2 expression also correlated with ESR and disease activity scores, such as of SLEDAI and BILAG, in SLE patients. Conclusion: The results indicate that essential amino acid methionine plays an important role in PB differentiation through the CD98-EZH2-axis. The pathological process of SLE seems to involve essential amino acids and their metabolic/activation pathways throughout the process of PB differentiation. Disclosure of Interests: Mingzeng Zhang: None declared, Shigeru Iwata: None declared, Maiko Hajime: None declared, Naoaki Ohkubo: None declared, Yasuyuki Todoroki: None declared, Hiroko Miyata: None declared, Shingo Nakayamada Grant/research support from: Mitsubishi-Tanabe, Takeda, Novartis and MSD, Speakers bureau: Bristol-Myers, Sanofi, Abbvie, Eisai, Eli Lilly, Chugai, Asahi-kasei and Pfizer, Yamagata Kaoru: None declared, Yoshiya Tanaka Grant/research support from: Abbvie, Astellas, Bristol-Myers Squibb, Chugai, Daiichi-Sankyo, Eisai, Mitsubishi-Tanabe, MSD, Ono, Taisho-Toyama, Takeda, Speakers bureau: Abbvie, Asahi-kasei, Astellas, Bristol-Myers Squibb, Chugai, Daiichi-Sankyo, Eli Lilly, Eisai, Glaxo-Smithkline, Janssen, Mitsubishi-Tanabe, Novartis, Pfizer Japan Inc, Sanofi, Takeda, UCB, YL Biologics

Background B cells play a crucial role in Systemic Lupus Erythematosus (SLE). Recently, ”Immunometabolism” attract much attention. Glucose and glutamine are important nutrition for energy production such as ATP in various cells. It has been reported that aerobic glycolysis, glutaminolysis and mitchondrial functions enhanced in cancer cells. However, the involvement of metabolic reprograming in plasmablast differentiation and its relevance to the pathogenesis of SLE remained elusive. Objectives We first investigated the abnormality of mitochondria in B cells from patients with SLE by flow cytometry. Next, B cells were isolated from healthy donors (HDs) and metabolic reprograming were assessed in vitro. Methods First, peripheral blood mononuclear cells (PBMCs) were obtained from age-matched 31 HDs and 29 patients with SLE. The mitochondrial membrane potential was measured with DiOc6 by flow cytometry. In addition, CD19+ cells were isolated from HDs and stimulated with CpG (TLR9 ligand) and IFN-α. Change of aerobic glycolysis, glutaminolysis and mitochondrial functions were assessed in the absence of glucose or glutamine and in the addition of metformine, which is known as AMPK activator, in vitro. Results We first examined the abnormality of mitochondria in B cells from patients with SLE using DiOc6 as a marker of depolarization-activated mitochondrial membrane. Baseline characteristics of SLE were males: females=1:28, age 40.2 years, disease duration 132.2 months, SLEDAI 6.4 and BILAG 6.2 at the timing of admission due to exacerbation of SLE. The percentage of IgD-CD27- memory B cells were higher than those of HDs, while the percentage of IgM memory B cells were lower than that of HDs. The percentage of CD24-DiOc6+ cells in IgD-CD27- memory B cells from patients with SLE were higher than that of HDs. These results indicate that B cells from patients with SLE have the abnormality of differentiation and mitochondrial functions. Next, we assessed the change of aerobic glycolysis, mitochondrial functions and glutaminolysis in the process of plasmablast differentiation in vitro. Stimulation with CpG and IFN-α, 1) enhanced aerobic glycolysis, which was assessed by lactate production. 2) increased the area of cytoplasm including many expanded mitochondrial cristae with slightly wider and loosely organized intermembrane space in electric microscopy, accompanied with ROS production and DiOc6 up-regulation, 3) induced CD27hiCD38hi plasmablasts differentiation and immunoglobulin production. Lactate production was decreased in the absence of both glucose or glutamine. ROS production and DiOc6 expression were decreased in the absence of glutamine, leading to inhibition of plasmablasts differentiation and immunoglobulin production. On the other hand, this tendency was not shown in the absence of glucose. Next, we evaluated oxygen consumption rate (OCR). OCR was also suppressed in the absence of glutamine. Metformin, abrogated glutamine uptake, resulting in suppression of ROS production, DiOc6 expression, plasmablast differentiation and immunoglobulin production. Conclusion These results suggest that mitochondrial activation via glutaminolysis may play an important role in the differentiation from IgD-CD27- double negative B cells to plasmablasts and production of immunoglobulins in patients with SLE. Disclosure of Interests Maiko Hajime: None declared, Shigeru Iwata: None declared, Mingzeng Zhang: None declared, Hiroko Miyata: None declared, SEUNGHYUN LEE: None declared, Shingo Nakayamada Grant/research support from: Mitsubishi-Tanabe, Takeda, Novartis and MSD, Speakers bureau: Bristol-Myers, Sanofi, Abbvie, Eisai, Eli Lilly, Chugai, Asahi-kasei and Pfizer, Kazuo Yamamoto: None declared, Yosuke Okada: None declared, Yoshiya Tanaka Grant/research support from: Abbvie, Astellas, Bristol-Myers Squibb, Chugai, Daiichi-Sankyo, Eisai, Mitsubishi-Tanabe, MSD, Ono, Taisho-Toyama, Takeda, Speakers bureau: Abbvie, Asahi-kasei, Astellas, Bristol-Myers Squibb, Chugai, Daiichi-Sankyo, Eli Lilly, Eisai, Glaxo-Smithkline, Janssen, Mitsubishi-Tanabe, Novartis, Pfizer Japan Inc, Sanofi, Takeda, UCB, YL Biologics

The aqueous solution equilibria of the systems containing copper(II) ion and glycinamide, β-alaninamide, glycine-N,N-diethylamide, or β-alanine-N,N-diethylamide have been investigated by potentiometric titration at 25°C (μ=0.1 (KNO3)). The stability constants K1 and K2 and the constants of the deprotonation reactions K1H, Kc1, and Kc2, for the glycinamide-copper(II) systems, and K1 and K2 for the other ligand-copper(II) systerrs were calculated by the method of non-linear least-squares with the aid of a computer. The K1 values were found to be in the order, glycine-N,N-diethylamide (6.17)>β-alanine-N,N-diethylamide (5.5)>glycinamide (5.30)>β-alaninamide (5.1). The sequence reveals that the five-membered chelates are more stable than the six-membered ones. The K1 and Kc1 values for glycylglycine and β-alanylglycine have been determined anew, and the structures of the chelates formed in acid solution have been discussed from comparative studies of the equilibrium constants, which suggest a tridentate nature...

Cardiovascular response to 2 min of isometric handgrip exercise at 50% of maximum voluntary contraction was studied echocardiographically in 10 essential hypertensives, before and during treatment with atenolol for a mean of 2 months. The patients responded with increases in heart rate, systolic and diastolic blood pressures, cardiac output and calculated triple product, no changes in stroke volume and total peripheral resistance, and decreases in ejection fraction, mean velocity of circumferential shortening and mean diastolic posterior wall velocity of the left ventricle before treatment. Chronic atenolol therapy attenuated the increases in heart rate, blood pressure and triple product, and the decreases in ejection fraction, mean velocity of circumferential shortening and mean diastolic posterior wall velocity of the left ventricle but resulted in a marked increase in total peripheral resistance. The pressure response and triple product rise in response to isometric handgrip exercise were also decreased. This suggests an obvious advantage to hypertensive patients who may, therefore, be protected from the risk of cardiovascular complications following isometric exercise.

Treatment of 2,4(5)-dialkyl-5(4)-phenylthioimidazoles with Raney nickel produced 2,4(5)-dialkylimidazoles. Methylation of 2,4(5)-dialkyl-5(4)-phenylthioimidazoles yielded 1-methyl-2,4-dialkylimidazoles, whereas methylation of 2,4(5)-dialkylimidazoles yielded in each case two isomers, the 1-methyl-2,4-dialkylimidazole and the 1-methyl-2,5-dialkylimidazole, which were not separated.