Fractalkine expression and the recruitment of CX3CR1+ cells in the prolonged mesangial proliferative glomerulonephritis

Fractalkine expression and the recruitment of CX3CR1+ cells in the prolonged mesangial proliferative glomerulonephritis.BackgroundWe established the reversible and the prolonged models of mesangial proliferative glomerulonephritis (GN) with anti-Thy 1 antibody 1-22-3. However, the essential factors leading to the prolonged glomerular alterations have not been identified.MethodsThe expressions of several chemokines and cytokines were compared in the reversible and the prolonged models. Expression of fractalkine and the number of the fractalkine receptor CX3CR1-positive cells in the glomeruli in the prolonged model were significantly higher than those in the reversible model. Then, the localization of fractalkine and the characteristics of CX3CR1+ cells were analyzed in glomeruli. To elucidate the significance of the fractalkine expression, we analyzed the expression in the model treated with angiotensin II receptor antagonist, candesartan.ResultsImmunostaining of fractalkine was detected on endothelial cells on the fifth day, and fractalkine staining also was detected in the mesangial area on day 14. Major parts of the CX3CR1+ cells in the glomeruli were macrophages, especially ED3+ cells. Candesartan treatment ameliorated the glomerular morphological findings at six weeks after disease induction. Although the treatment did not ameliorate the morphological finding at two weeks, decreased expression of fractalkine and CX3CR1+ were already detected at two weeks in rats treated with candesartan.ConclusionsFractalkine expression and the recruitment of CX3CR1+ cells in glomeruli might play an important role in the development of the prolonged disease. These expressions could be predictors of the prolonged disease of the mesangial proliferative glomerulonephritis

Insight into innate immune response in “Yusho”: The impact of natural killer cell and regulatory T cell on inflammatory prone diathesis of Yusho patients

Background: In 1968 in western Japan, polychlorinated biphenyl-contaminated “Kanemi rice oil” was used in cooking, causing food poisoning in many people. More than 50 years have passed since the Yusho incident, and although inflammatory disorders such as suppuration have been observed in Yusho patients, the etiology of this inflammation susceptibility remains obscure. Objectives: To investigate the mechanisms of susceptibility to inflammation in Yusho patients, peripheral immune cell fractions and concentrations of inflammatory cytokines were evaluated in blood samples collected from both Yusho patients and age-matched healthy subjects undergoing medical examination in Nagasaki. Methods: To exclude diagnostic uncertainty, serum levels of polychlorinated biphenyl (PCB), polychlorinated quarterphenyl (PCQ), and polychlorinated dibenzofuran (PCDF) were measured. Immune cell (e.g. natural killer and regulatory T cell) populations were analyzed by flow cytometry. Serum cytokines involved in immune cell activation were measured by ELISA. Results: The relative proportion of natural killer cells was higher in Yusho patients than in healthy subjects, while the proportion of regulatory T cells did not differ between groups. Serum concentrations of IL-36 and IFN-γ were significantly lower in Yusho patients than in healthy subjects. Conversely, serum cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), which is a cytokine related to activated NK cells, was higher in Yusho patients than in healthy subjects and was positively correlated with PCDF blood levels. Conclusion: Increased numbers of NK cells in Yusho patients suggests that the innate immune response has been activated in Yusho patients. The seemingly paradoxical results for CTLA-4 and IFN-γ may reflect counterbalancing mechanisms preventing excessive NK cell activation. This dysregulation of innate immunity might contribute to the inflammation observed in Yusho patients

Minor contribution of CYP3A5 to the metabolism of hepatitis C protease inhibitor paritaprevir in vitro

Paritaprevir (PTV) is a non-structural protein 3/4A protease inhibitor developed for the treatment of hepatitis C disease as a fixed dose combination of ombitasvir (OBV) and ritonavir (RTV) with or without dasabuvir. The aim of this study was to evaluate the effects of cytochrome P450 (CYP) 3A5 on in vitro PTV metabolism using human recombinant CYP3A4, CYP3A5 (rCYP3A4, rCYP3A5) and human liver microsomes (HLMs) genotyped as either CYP3A5*1/*1, CYP3A5*1/*3 or CYP3A5*3/*3. The intrinsic clearance (CLint, Vmax/Km) for the production of a metabolite from PTV in rCYP3A4 was 1.5 times higher than that in rCYP3A5. The PTV metabolism in CYP3A5*1/*1 and CYP3A5*1/*3 HLMs expressing CYP3A5 was comparable to that in CYP3A5*3/*3 HLMs, which lack CYP3A5. CYP3A4 expression level was significantly correlated with PTV disappearance rate and metabolite formation. In contrast, there was no such correlation found for CYP3A5 expression level. This study represents that the major CYP isoform involved in PTV metabolism is CYP3A4, with CYP3A5 having a minor role in PTV metabolism. The findings of the present study may provide foundational information on PTV metabolism, and may further support dosing practices in HCV-infected patients prescribed PTV-based therapy

Chondroitin sulfate N-acetylgalactosaminyltransferase-1 is required for normal cartilage development

CS (chondroitin sulfate) is a glycosaminoglycan species that is widely distributed in the extracellular matrix. To understand the physiological roles of enzymes involved in CS synthesis, we produced CSGalNAcT1 (CS N-acetylgalactosaminyltransferase 1)-null mice. CS production was reduced by approximately half in CSGalNAcT1-null mice, and the amount of short-chain CS was also reduced. Moreover, the cartilage of the null mice was significantly smaller than that of wild-type mice. Additionally, type-II collagen fibres in developing cartilage were abnormally aggregated and disarranged in the homozygous mutant mice. These results suggest that CSGalNAcT1 is required for normal CS production in developing cartilage

Clinical Study of Pollen-food Allergy Syndrome Estimated by Double-blind, Placebo-controlled Food Challenges of Ten Apple Cultivars

Background : The number of patients with pollen-food allergy syndrome (PFAS) has increased globally. Apples are a major causative food for PFAS. Varied reports have demonstrated the usefulness of skin prick test (SPT) for the diagnosis of PFAS, besides differences in symptom expression among apple cultivars, thus the diagnosis of PFAS remains unclear. Purpose : To investigate the clinical features of apple-induced PFAS by performing a double-blind placebocontrolled food challenge (DBPCFC) using different apple cultivars on patients with apple-induced PFAS. Method : DBPCFC was performed for six patients with apple-induced PFAS using 10 apple cultivars. We measured the degree of symptoms using the Visual Analog Scale (VAS). Further, we assessed the correlations of Mal d 1 and Bet v 1-specific IgE levels and SPT findings to the VAS. Results : Three of six patients (50 ％) were VAS-positive for two apple cultivars, one patient each (17 ％) was positive for three, four, and five apple cultivars. The correlation between SPT findings and VAS was insignificant (p＝0.103). The VAS displayed a positive relationship with Mal d 1 and Bet v 1 (r＝0.5 and r＝0.84, respectively). Conclusion : It is necessary to perform DBPCFC with multiple apple cultivars to diagnose PFAS. The SPT was not useful in diagnosing PFAS ; however, Bet v 1-specific IgE levels may be advantageous. This novel clinical study of PFAS assessed multiple Japanese apple varieties, therefore, our findings can serve as a baseline for future studies.Article信州医学雑誌 71(2) : 99-107, (2023)journal articl

Clinical Study of Pollen-food Allergy Syndrome Estimated by Double-blind, Placebo-controlled Food Challenges of Ten Apple Cultivars

Background : The number of patients with pollen-food allergy syndrome (PFAS) has increased globally. Apples are a major causative food for PFAS. Varied reports have demonstrated the usefulness of skin prick test (SPT) for the diagnosis of PFAS, besides differences in symptom expression among apple cultivars, thus the diagnosis of PFAS remains unclear. Purpose : To investigate the clinical features of apple-induced PFAS by performing a double-blind placebocontrolled food challenge (DBPCFC) using different apple cultivars on patients with apple-induced PFAS. Method : DBPCFC was performed for six patients with apple-induced PFAS using 10 apple cultivars. We measured the degree of symptoms using the Visual Analog Scale (VAS). Further, we assessed the correlations of Mal d 1 and Bet v 1-specific IgE levels and SPT findings to the VAS. Results : Three of six patients (50 ％) were VAS-positive for two apple cultivars, one patient each (17 ％) was positive for three, four, and five apple cultivars. The correlation between SPT findings and VAS was insignificant (p＝0.103). The VAS displayed a positive relationship with Mal d 1 and Bet v 1 (r＝0.5 and r＝0.84, respectively). Conclusion : It is necessary to perform DBPCFC with multiple apple cultivars to diagnose PFAS. The SPT was not useful in diagnosing PFAS ; however, Bet v 1-specific IgE levels may be advantageous. This novel clinical study of PFAS assessed multiple Japanese apple varieties, therefore, our findings can serve as a baseline for future studies.Article信州医学雑誌 71(2) : 99-107, (2023