Interleukins IL-4 and IL-10 are considered to be central regulators for the limitation and eventual termination of inflammatory responses in vivo, based on their potent anti-inflammatory effects toward LPS-stimulated monocytes/macrophages and neutrophils. However, their role in T cell-dependent inflammatory responses has not been fully elucidated. In this study, we investigated the effects of both cytokines on the production of PGE(2), a key molecule of various inflammatory conditions, in CD40-stimulated human peripheral blood monocytes. CD40 ligation of monocytes induced the synthesis of a significant amount of PGE(2) via inducible expression of the cyclooxygenase (COX)-2 gene. Both IL-10 and IL-4 significantly inhibited PGE(2) production and COX-2 expression in CD40-stimulated monocytes. Using specific inhibitors for extracellular signal-related kinase (ERK) and p38 mitogen-activated protein kinase (MAPK), we found that both kinase pathways are involved in CD40-induced COX-2 expression. CD40 ligation also resulted in the activation of NF-kappaB. Additional experiments exhibited that CD40 clearly induced the activation of the upstream kinases MAPK/ERK kinase 1/2, MAPK kinase 3/6, and I-kappaB in monocytes. IL-10 significantly inhibited CD40-induced activation of the ERK, p38 MAPK, and NF-kappaB pathways; however, inhibition by IL-4 was limited to the ERK pathway in monocytes. Neither IL-10 nor IL-4 affected the recruitment of TNFR-associated factors 2 and 3 to CD40 in monocytes. Collectively, IL-10 and IL-4 use novel regulatory mechanisms for CD40-induced prostanoid synthesis in monocytes, thus suggesting a potential role for these cytokines in regulating T cell-induced inflammatory responses, including autoimmune diseases.
Abstract Objective To evaluate trends in results of care and management for antineutrophil cytoplasmic antibodies (ANCA)‐associated vasculitis (AAV). Methods We employed multicenter cohort data collected during 2011–2021, recruiting 43 patients with granulomatosis with polyangiitis (GPA) and 91 with microscopic polyangiitis (MPA). According to the median registration date of September 2015, patients have split into two groups: an early group and a late group (both of them, n = 67). To prevent bias, a propensity score according to numerous baseline characteristics variables was calculated; 50 matching members of each group were statistically extracted. Their treatments and clinical outcomes were examined at 6, 12, and 24 months after initial remission therapy. Results Statistics demonstrated that the baseline characteristics were similar. The late group used rituximab (RTX) more often for both remission induction and maintenance therapy, compared with the early group. The mean daily PSL doses of the late group were significantly lower than those of early group at each time point. The late group discontinued PSL 14.0% at 12 months and 23.3% at 24 months. Despite their intensive glucocorticoids (GC) tapering, the remission rates and the relapse rates were significantly fairer in the late group. The Vasculitis Damage Index (VDI) and VDI due to GC at each time point were lower in the late group, and those differences had become wider over time. Conclusion Recent developments in AAV treatment have allowed efficient remission and prevention of relapses, which in turn enabled extensive GC tapering causing fewer sequelae.
Abstract A synthesis of acyl halides from the corresponding carboxylic acid thioesters was achieved using commercially available Selectfluor or NCS, and the acyl fluoride or chloride intermediates were transformed to the corresponding esters, amides, and several carbon‐carbon bond formation products. This approach can be applied to the peptide synthesis from functionalized amino acid thioesters.
Abstract Background The pepsin agglutinator, discovered over 50 years ago, has recently referred to be an anti-hinge antibody (AHA) because of main reacting with the IgG hinge epitope. AHA shows different reactivity for each hinge epitope generated by each protease that cleaves the hinge region at different sites. Moreover, AHA shows different reactivity against different hinge epitopes derived from each IgG subclass even when the same protease is used. Since production of matrix metalloproteinase-3 (MMP-3) is enhanced in rheumatoid arthritis (RA), the specific AHA might be increasingly produced. The purpose of this study is to determine whether AHA against IgG hinge epitopes produced by MMP-3 is specifically elevated in RA. Methods Serum IgG or IgA class of AHA against the IgG1- or IgG4 F(ab’) 2 generated by either matrix metalloproteinase 3 (MMP-3) or pepsin was measured by ELISA in 81 healthy controls (HC) and 111 patients with RA. Receiver operating characteristic (ROC) analysis was used for obtaining optimal cutoff values and cutoff values indicating high specificity (>95%) of the AHA. Targeted epitope of the AHA was investigated by inhibition ELISA. Results Seven AHAs were statistically higher in RA than HC, except IgG AHA against IgG1 F(ab’) 2 generated by proteolytic cleavage of MMP-3. The areas under the curve of ROC curve were 0.66-0.80, although the sensitivities at high specificity were low (5.4-24.3%). The cumulative number of positive AHA in each individual was statistically higher in RA than HC, suggesting the extreme extent of AHA repertoires in RA. Inhibition studies revealed that IgG AHA against IgG4 F(ab’) 2 generated by pepsin cross-reacted with IgG1 F(ab’) 2 generated by pepsin. Although development of IgA AHAs in RA seems to be remarkable and specific, multivariate logistic regression analysis identified IgG AHA against IgG4 F(ab’) 2 generated by pepsin as an independent variable for RA diagnosis, even in RA patients who were negative for both RF and ACPA [odds ratio 1.18 (95% confidence interval 1.06-1.32); P =0.003]. Conclusion In RA, we observed diversification and amplification of AHA repertoires and diagnostic utility of the specific AHA against IgG4 F(ab’) 2 generated by pepsin, but not MMP-3.
Abstract Background. Pepsin agglutinators, discovered over 50 years ago, have been recently referred to as anti-hinge antibodies (AHAs) because of their reaction with the IgG hinge epitope. AHAs have different reactivity for each hinge epitope generated by each protease that cleaves the hinge region at different sites. Moreover, AHAs have different reactivity against different hinge epitopes derived from each IgG subclass even when the same protease is used. Since the expression of matrix metalloproteinase-3 (MMP-3) is enhanced in rheumatoid arthritis (RA), AHA production could also be increased. The purpose of this study was to determine whether the levels of AHAs against IgG hinge epitopes produced by MMP-3 are elevated in RA. Methods. The serum levels of IgG or IgA AHAs against the IgG1/IgG4 F(ab’) 2 fragments, generated by either MMP-3 or pepsin, was measured using ELISA in 111 patients with RA and 81 healthy controls (HC). Receiver operating characteristic (ROC) analysis was used for obtaining optimal cutoff values and cutoff values indicating high specificity (>95%) of the AHA. The targeted epitope of a specific AHA was investigated through inhibition ELISA. Results. Seven AHAs were statistically higher in RA patients than HC, except IgG AHA against IgG1 F(ab’) 2 , which was generated by MMP-3 proteolytic cleavage. The areas under the ROC curve were of 0.66--0.80, although the sensitivities at high specificity were low (5.4--24.3%). The cumulative number of positive AHAs in each individual was statistically higher in RA patients than HC, suggesting the extreme extent of AHA repertoires in RA. Inhibition studies revealed that IgG AHAs against IgG4 F(ab’) 2 fragments generated by pepsin cross-reacted with IgG1 F(ab’) 2 fragments generated by pepsin. Multivariate logistic regression analysis identified the IgG AHA against IgG4 F(ab’) 2 fragments generated by pepsin as an independent variable for RA diagnosis, even in RA patients who were negative for both RF and ACPA (odds ratio 1.18, 95% CI: 1.06−1.32; P =0.003). Additional experiments using non-RA patients finally strengthened the diagnostic utility. Conclusion. In RA patients, we observed diversification and amplification of AHA repertoires and diagnostic utility of the specific AHA against IgG4 F(ab’) 2 fragments generated by pepsin but not MMP-3.
TNF inhibitors are recommended as the first line biologics to date because of a great deal of evidence, however, some patients show more obvious response to IL-6 inhibitor (tocilizumab; TCZ) treatment. For the purpose of achieving early remission and efficient medical economics, it would be desirable to be able to predict the efficacy of each anti-cytokine therapy before treatment.
Objectives
The purpose of this study is to establish a scoring method that predicts preferable treatment before starting either TCZ or TNF inhibitor.
Methods
First, mRNA levels of IL-6 and TNF-α in the peripheral blood of 45 newly diagnosed RA patients were analyzed by using Agilent whole human genome 60K cDNA arrays. The correlation analysis between mRNA levels of IL-6 and TNF-α and the correlation analysis between each laboratory datum and the difference between IL-6 and TNF-α mRNA levels were performed. Next, 183 patients given anti-cytokine therapy in our hospital were retrospectively analyzed. They were divided into 2 groups, Group A consisted of 37 patients treated with TCZ, and Group B consisted of 146 patients treated with TNF inhibitor. The correlation analysis between the labo-data at baseline and the ratio of the DAS28-ESR before and 6 months after treatment (DAS ratio) was performed for each group. Labo-data with a significant correlation to the DAS ratio were selected, and a scoring formula for predicting the efficacy of TCZ treatment was devised by using those labo-data. On the basis of this formula, a scoring table was devised. The propriety of this scoring system was investigated by the clinical data of 183 RA patients.
Results
The correlation analysis between mRNA levels of IL-6 and TNF-α showed a significant inverse correlation between them. In order to find the useful laboratory datum to represent the balance between IL-6 and TNF-α, the correlation analysis was performed between each laboratory datum and the difference between IL-6 and TNF-α mRNA levels. It showed that hemoglobin (Hb), platelet count (Plt), ALT, and ESR have significant correlation to the difference of IL-6 and TNF-α level. AST also have weak correlation to it. Next, the correlation analysis between DAS ratio and each labo-data in Group A showed that Hb, Plt, AST and ALT had significant correlation to the DAS ratio, whereas no similar correlation was shown in the Group B. On the basis of these results, the following scoring formula was devised: Score = {120 × Plt1.4 × Hb(–1.9) × AST(–0.65) × ALT(–0.65)}2, Then, on the basis of above formula, a scoring table was devised. Each item (Plt, Hb, AST, and ALT) has 0-10 points, and the full score is 40 points. When the scores calculated by this table were more than 20 points, response rates of Group A vs Group B were as follows; good resonse 60% vs 0%, more than moderate response 100% vs 66.7%, No response 0% vs 33.3%.
Conclusions
The finding of inverse correlation between mRNA expression of IL-6 and of TNF-α in RA supports a possibility to establish a scoring method to predict dominant cytokine that should be targeted for the treatment. As our scoring system was devised from retrospective analysis, it requires accumulation of data for the better accuracy. However this kind of approach would help to guide the selection of either the TNF inhibitor or the IL-6 inhibitor for each RA patient.
To compare therapeutic efficacy of tumour necrosis factor inhibitor (TNFi) cyclers and non-TNFi switchers in patients with rheumatoid arthritis (RA) having inadequate response to previous TNFis (TNF-IR patients) using composite measures including imaging assessment with power Doppler ultrasonography (PDUS). Patients with RA who had inadequate response to one or more previous TNFi agents with moderate or higher disease activity were enrolled. The outcomes of 56 TNF-IR patients were analysed. Patients were divided into 19 TNFi cyclers and 37 non-TNFi switchers (16 abatacept [ABT] and 21 tocilizumab [TCZ] switchers). Retention ratio at 6 months was significantly higher in non-TNFi switchers than in TNFi cyclers (p < .05). Although there was no significant difference, non-TNFi switchers tended to have a larger decrease than TNFi cyclers in efficacy indicators based on clinical disease activity index and PDUS. Multivariate logistic regression analysis identified a following independent factor associated with both EULAR good response and retention of a biologic agent: non-TNFi switch (p < .05 for both). Non-TNFi switchers were shown to have significantly higher percentage of EULAR good response and higher retention than TNFi cyclers. A non-TNFi biologic agent may hence be a preferential next-line treatment for TNF-IR patients.
Cap layers for Cu interconnects in ultra-large-scale integrated devices (ULSIs), with a low dielectric constant (k-value) and strong barrier properties against Cu and moisture diffusion, are required for the future further scaling of ULSIs. There is a trade-off, however, between reducing the k-value and maintaining strong barrier properties. Using quantum mechanical simulations and other theoretical computations, we have designed ideal dielectrics: SiCH films with Si-C2H4-Si networks. Such films were estimated to have low porosity and low k; thus they are the key to realizing a cap layer with a low k and strong barrier properties against diffusion. For fabricating these ideal SiCH films, we designed four novel precursors: isobutyl trimethylsilane, diisobutyl dimethylsilane, 1, 1-divinylsilacyclopentane and 5-silaspiro [4,4] noname, based on quantum chemical calculations, because such fabrication is difficult by controlling only the process conditions in plasma-enhanced chemical vapor deposition (PECVD) using conventional precursors. We demonstrated that SiCH films prepared using these newly designed precursors had large amounts of Si-C2H4-Si networks and strong barrier properties. The pore structure of these films was then analyzed by positron annihilation spectroscopy, revealing that these SiCH films actually had low porosity, as we designed. These results validate our material and precursor design concepts for developing a PECVD process capable of fabricating a low-k cap layer.
