Showing papers by "Kenji Matsumoto published in 2020"

Innate Lymphoid Cells in the Airways: Their Functions and Regulators.

Abstract: Since the airways are constantly exposed to various pathogens and foreign antigens, various kinds of cells in the airways-including structural cells and immune cells-interact to form a precise defense system against pathogens and antigens that involve both innate immunity and acquired immunity. Accumulating evidence suggests that innate lymphoid cells (ILCs) play critical roles in the maintenance of tissue homeostasis, defense against pathogens and the pathogenesis of inflammatory diseases, especially at body surface mucosal sites such as the airways. ILCs are activated mainly by cytokines, lipid mediators and neuropeptides that are produced by surrounding cells, and they produce large amounts of cytokines that result in inflammation. In addition, ILCs can change their phenotype in response to stimuli from surrounding cells, which enables them to respond promptly to microenvironmental changes. ILCs exhibit substantial heterogeneity, with different phenotypes and functions depending on the organ and type of inflammation, presumably because of differences in microenvironments. Thus, ILCs may be a sensitive detector of microenvironmental changes, and analysis of their phenotype and function at local sites may enable us to better understand the microenvironment in airway diseases. In this review, we aimed to identify molecules that either positively or negatively influence the function and/or plasticity of ILCs and the sources of the molecules in the airways in order to examine the pathophysiology of airway inflammatory diseases and facilitate the issues to be solved.

Cumulative inactivated vaccine exposure and allergy development among children: A birth cohort from Japan

Abstract: Adjuvants used in inactivated vaccines often upregulate type 2 immunity, which is dominant in allergic diseases. We hypothesised that cumulative adjuvant exposure in infancy may influence the development of allergies later in life by changing the balance of type 1/type 2 immunity. We examined the relationship between immunisation with different vaccine types and later allergic disease development. We obtained information regarding vaccinations and allergic diseases through questionnaires that were used in The Japan Environment and Children’s Study (JECS), which is a nationwide, multicentre, prospective birth cohort study that included 103,099 pregnant women and their children. We examined potential associations between the initial vaccination before 6 months of age and symptoms related to allergies at 12 months of age. Our statistical analyses included 56,277 children. Physician-diagnosed asthma was associated with receiving three (aOR 1.395, 95% CI 1.028–1.893) or four to five different inactivated vaccines (aOR 1.544, 95% CI 1.149–2.075), compared with children who received only one inactivated vaccine. Similar results were found for two questionnaire-based symptoms, i.e. wheeze (aOR 1.238, 95% CI 1.094–1.401; three vaccines vs. a single vaccine) and eczema (aOR 1.144, 95% CI 1.007–1.299; four or five vaccines vs. a single vaccine). Our results, which should be cautiously interpreted, suggest that the prevalence of asthma, wheeze and eczema among children at 12 months of age might be related to the amount of inactivated vaccine exposure before 6 months of age. Future work should assess if this association is due to cumulative adjuvant exposure. Despite this possible association, we strongly support the global vaccination strategy and recommend that immunisations continue. UMIN000030786 .

Robust production of IL-33 and TSLP by lung endothelial cells in response to low-dose dsRNA stimulation

Abstract: Compared to other airway tissue cells (e.g., bronchial epithelial cells), lung vascular endothelial cells robustly produced both IL-33 and TSLP proteins, which are believed to be epithelium-derived cytokines, in response to dsRNA stimulation. (33 words).

Functional benefits of corticosteroid and IVIG combination therapy in a coronary artery endothelial cell model of Kawasaki disease.

Abstract: Kawasaki disease (KD) is the most common pediatric systemic vasculitides of unknown etiology. Recent clinical studies led to reappraisal of the usefulness of initial combination therapy of intravenous immunoglobulin (IVIG) plus a corticosteroid for patients with severe KD. However, the molecular mechanisms underlying the clinical benefits of that combination therapy remain unclear. Here, we used cultured human coronary artery endothelial cells (HCAECs), as a mimic of KD, to study the possible mechanisms responsible for the clinical benefits of adding a corticosteroid to standard IVIG therapy for patients with severe KD. HCAECs were stimulated with TNF-α, IL-1α or IL-1β in the presence and absence of high-dose IgG and/or dexamethasone (DEX). The mRNA and protein concentrations for high-mobility group box-1 (HMGB1), IL-1α, IL-6 and granulocyte-colony stimulating factor (G-CSF) in the culture supernatants were measured by quantitative PCR (qPCR) and ELISA, respectively. Apoptosis was evaluated by the caspase 3/7 activities. DEX, but not IgG, significantly inhibited apoptosis caused by inflammatory stimuli, resulting in effective reduction of HMGB1 and IL-1α protein release by HCAECs. As previously reported, DEX or IgG alone significantly suppressed TNF-α-induced production of IL-6 and G-CSF and mRNA expression, but induction of those cytokines by IL-1 s (IL-1α and IL-1β) was resistant to high-dose IgG. A corticosteroid can effectively inhibit the release of HMGB1 and IL-1α, which may be involved in IVIG resistance in KD. Since high-dose IgG does not have such beneficial anti-cytotoxic effects, adding a corticosteroid to standard IVIG therapy may help prevent the progression of IVIG resistance in KD.

IL-33 Is Essential for Adjuvant Effect of Hydroxypropyl-β-Cyclodexrin on the Protective Intranasal Influenza Vaccination.

Abstract: Vaccine adjuvants are traditionally used to augment and modulate the immunogenicity of vaccines, although in many cases it is unclear which specific molecules contribute to their stimulatory activity. We previously reported that both subcutaneous and intranasal administration of hydroxypropyl-β-cyclodextrin (HP-β-CD), a pharmaceutical excipient widely used to improve solubility, can act as an effective adjuvant for an influenza vaccine. However, the mechanisms by which mucosal immune pathway is critical for the intranasal adjuvant activity of HP-β-CD have not been fully delineated. Here, we show that intranasally administered HP-β-CD elicits a temporary release of IL-33 from alveolar epithelial type 2 cells in the lung; notably, IL-33 expression in these cells is not stimulated following the use of other vaccine adjuvants. The experiments using gene deficient mice suggested that IL-33/ST2 signaling is solely responsible for the adjuvant effect of HP-β-CD when it is administered intranasally. In contrast, the subcutaneous injection of HP-β-CD and the intranasal administration of alum, as a damage-associated molecular patterns (DAMPs)-inducing adjuvant, or cholera toxin, as a mucosal adjuvant, enhanced humoral immunity in an IL-33-independent manner, suggesting that the IL-33/ST2 pathway is unique to the adjuvanticity of intranasally administered HP-β-CD. Furthermore, the release of IL-33 was involved in the protective immunity against influenza virus infection which is induced by the intranasal administration of HP-β-CD-adjuvanted influenza split vaccine. In conclusion, our results suggest that an understanding of administration route- and tissue-specific immune responses is crucial for the design of unique vaccine adjuvants.

IL-1β and IL-17A are involved in IVIG resistance through activation of C/EBPβ and δ in a coronary artery model of Kawasaki disease.

Abstract: To the Editor, Kawasaki disease (KD) is a pediatric systemic vasculitis of unknown etiology. Intravenous immunoglobulin (IVIG) has been the standard treatment for KD and is highly effective in almost 80% of patients. However, the remaining patients are resistant to initial IVIG treatment and are at high risk for coronary artery lesions (CALs) due to severe coronary inflammation. That resistance is the most important issue needing to be clinically resolved for KD. Therefore, there is an urgent need for the development of novel therapies for IVIGresistant KD based on an understanding of the mechanism of that resistance. We show here that activation of two transcription factors, CCAAT/enhancer-binding protein beta and delta (C/EBPβ and C/EBPδ), whose target genes include IL-6 and G-CSF, plays a crucial role in IVIG-resistant inflammation of human coronary artery endothelial cells (HCAECs). Accumulating evidence suggests that both IL-1β and IL-17A are crucially involved in IVIG resistance in KD patients.1-6 We previously demonstrated that IVIG could specifically and completely suppress TNF-α-induced, but not IL-1β-induced, protein production of such KD-related cytokines as IL-6 and G-CSF by HCAECs.7 Furthermore, the dramatic and specific anti-inflammatory effects of IVIG were associated with complete inhibition of C/EBP activity. In this study, we aimed to elucidate the effects of IL-1β and IL-17A on the anti-inflammatory effects of IVIG and the involvement of C/EBPβ and C/EBPδ in those effects. The detailed methods are provided in Appendix S1. Consistent with our previous study, IVIG completely inhibited TNF-α-induced production of IL-6 and G-CSF by HCAECs. In contrast, we newly found that the striking anti-inflammatory effects of IVIG were almost completely abolished by the addition of IL-1β to TNF-α even at a ratio of only 1:100 (Figure 1A, upper panels). Similar to IL-1β, adding IL-17A to TNF-α attenuated IVIG's anti-inflammatory effects, but the effect was weaker than that of IL-1β (Figure 1A, lower panels). Indeed, IL-17A must be present in at least a 1:1 ratio to TNF-α. The mRNA expression patterns for both IL-6 and G-CSF (Figure 1B) were in line with their protein production patterns (Figure 1A). Since the levels of IL-6 and G-CSF production induced by combined TNF-α and IL-1β stimulation (Figure 1A, upper panels) and those induced by IL-1β stimulation alone (Figure S1A) were almost the same, it was clear that TNF-α had no additive effect on IL-1β stimulation of HCAECs. Therefore, the Western blot (WB) and electrophoretic mobility shift assay (EMSA) experiments shown in Figure 2 were conducted using only IL-1β stimulation. IVIG treatment completely suppressed TNF-α–induced nuclear expression of C/EBPβ and C/EBPδ (Figure 2A; WB) and their DNA-binding activity (Figure 2B; EMSA). Conversely, those effects were abolished when the cells were stimulated with IL-1β alone or a combination of TNF-α and IL-17A (Figure 2). These results strongly suggest that C/EBPβ and C/EBPδ activities elicited by IL-1β or IL-17A could be crucially involved in attenuation of IVIG's anti-inflammatory effect on HCAECs. To confirm the involvement of C/EBPs in cytokine-induced expression of IL-6 and G-CSF by HCAECs, HCAECs were transfected with siRNAs targeting C/EBPβ and C/EBPδ. Transfection of siRNAs for C/EBPβ and C/EBPδ into HCAECs successfully suppressed their respective mRNAs (Figure S2A). siRNAs for each of C/EBPβ and C/EBPδ independently suppressed IL-6 and G-CSF production by HCAECs. Moreover, co-transfection with siRNAs for both C/EBPβ and C/EBPδ showed a tendency for synergistic inhibition, especially upon IL-1β stimulation (Figure S2B). These results suggest that both C/EBPβ and C/EBPδ are required for cytokine-induced IL-6 and G-CSF production by HCAECs. 4. Dewald G. A missense mutation in the plasminogen gene, within the plasminogen kringle 3 domain, in hereditary angioedema with normal C1 inhibitor. Biochem Biophys Res Commun. 2018;498:193-198. 5. Ghannam A, Sellier P, Defendi F, et al. C1 inhibitor function using contact-phase proteases as target: evaluation of an innovative assay. Allergy. 2015;70:1103-1111. 6. Defendi F, Charignon D, Ghannam A, et al. Enzymatic assays for the diagnosis of bradykinin-dependent angioedema. PLoS ONE. 2013;8:e70140. 7. Baroso R, Sellier P, Defendi F, et al. Kininogen cleavage assay: diagnostic assistance for kinin-mediated angioedema conditions. PLoS ONE. 2016;11:e0163958. 8. Takada A, Takada Y. The activation of two isozymes of glu-plasminogen (I and II) by urokinase and streptokinase. Thromb Res. 1983;30:633-642. 9. Gonzalez-Gronow M, Gawdi G, Pizzo SV. Tissue factor is the receptor for plasminogen type 1 on 1-LN human prostate cancer cells. Blood. 2002;99:4562-4567.

Conjunctival Injection Reduction in Patients with Atopic Keratoconjunctivitis Due to Synergic Effect of Bovine Enteric-Coated Lactoferrin in 0.1% Tacrolimus Ophthalmic Suspension.

Abstract: Lactoferrin (LF), a multifunctional glycoprotein found in mammalian milk, is reported to have immunoregulatory effects. The present study aimed to evaluate whether enteric-coated LF (eLF) could improve symptoms in patients with atopic keratoconjunctivitis (AKC). This randomized double-blind placebo-controlled single-center trial comprised Japanese patients (n = 20; aged 22–60 years) with AKC. Patients treated with 0.1% tacrolimus ophthalmic suspension (TALYMUS®) were administered eLF (400 mg/d of bovine LF) or placebo tablets for 12 weeks. Conjunctival injection was examined, papillae formation in the palpebral conjunctiva was evaluated, and corneal fluorescein score, itchy sensation in end-point itching scale, and serum allergic parameters were assessed. Conjunctival injection was significantly reduced in the LF group than in the placebo group (p = 0.0017, Mann–Whitney U-test). Papillae formation in the palpebral conjunctiva showed a statistical decrease in the LF group than in the placebo group (p = 0.010, unpaired T-test). LF combined with TALYMUS® could be a promising treatment strategy to mitigate AKC.

Podocyte-specific deletion of tubular sclerosis complex 2 promotes focal segmental glomerulosclerosis and progressive renal failure

Abstract: Obesity can initiate and accelerate the progression of kidney diseases. However, it remains unclear how obesity affects renal dysfunction. Here, we show that a newly generated podocyte-specific tubular sclerosis complex 2 (Tsc2) knockout mouse model (Tsc2Δpodocyte) develops proteinuria and dies due to end-stage renal dysfunction by 10 weeks of age. Tsc2Δpodocyte mice exhibit an increased glomerular size and focal segmental glomerulosclerosis, including podocyte foot process effacement, mesangial sclerosis and proteinaceous casts. Podocytes isolated from Tsc2Δpodocyte mice show nuclear factor, erythroid derived 2, like 2-mediated increased oxidative stress response on microarray analysis and their autophagic activity is lowered through the mammalian target of rapamycin (mTOR)—unc-51-like kinase 1 pathway. Rapamycin attenuated podocyte dysfunction and extends survival in Tsc2Δpodocyte mice. Additionally, mTOR complex 1 (mTORC1) activity is increased in podocytes of renal biopsy specimens obtained from obese patients with chronic kidney disease. Our work shows that mTORC1 hyperactivation in podocytes leads to severe renal dysfunction and that inhibition of mTORC1 activity in podocytes could be a key therapeutic target for obesity-related kidney diseases.

Gene expression and DNA methylation changes in BeWo cells dependent on tumor necrosis factor-α and insulin-like growth factor-I.

Abstract: Pregnant women with increased insulin resistance, characterized by elevated levels of tumor necrosis factor-alpha (TNF-α) and insulin-like growth factor-I (IGF-I), are at high risk of preeclampsia. We hypothesized that TNF-α and IGF-I affect the placentas and cause pathological changes leading to preeclampsia. To understand the genetic and epigenetic effects of TNF-α and IGF-I on trophoblast cells, gene expression microarray and DNA methylation array of BeWo cells stimulated by TNF-α (100 pg/ml, 100 ng/ml) and IGF-I (100 ng/ml) were conducted. Microarray analysis revealed the differential gene expression patterns in BeWo cells co-stimulated by TNF-α and IGF-I. Enrichment analysis identified the terms associated with NF-kappa B signaling pathways and arachidonic acid cascades such as PTGS2 and PTGER2. DNA methylation array revealed the distinct CpG methylation pattern in BeWo cells stimulated by high-TNF-α and IGF-I, while neither of them showed independent effects. Enrichment analysis identified the terms associated with major histocompatibility complex proteins. Integration of transcriptome and DNA methylome analyses identified three differentially expressed genes with significant DNA methylation change: C3, GP1BA, and NFKBIE, which are all possibly associated with pathogenesis of preeclampsia. In conclusion, co-stimulation of TNF-α and IGF-I induced the genetic and epigenetic changes associated with preeclampsia in BeWo cells. The results suggested that BeWo cells stimulated by TNF-α and IGF-I is a good in vitro model of preeclamptic placenta in pregnancy with increased insulin resistance.

The Japanese experience and pharmacokinetics of antenatal maternal high-dose immunoglobulin treatment as a prophylaxis for neonatal hemochromatosis in siblings

Abstract: Background: Neonatal hemochromatosis (NH) is a rare but serious disease causing fulminant hepatic failure. The recurrence rate of NH in a subsequent infant of a mother with an affected infant is 70...

Effect of Transcatheter Aortic Valve Implantation on the Immune Response Associated With Surgical Aortic Valve Replacement

Abstract: The immune response after transcatheter aortic valve implantation (TAVI) in comparison to that after surgical aortic valve replacement (SAVR) remains to be fully elucidated. In a 2-part study, we assessed laboratory data obtained before, immediately after, and 24 and 48 hours after SAVR (128 patients; age ≥80 [mean 82] years) or transfemoral TAVI (102 patients; age ≥80 [mean 86] years) performed for aortic stenosis. In-hospital mortalities were similar (3% vs 0%), but leukocyte counts and aspartate aminotransferase and creatine kinas concentrations were decreased immediately and 24 hours after surgery (all, p 1.7 and p