scholarly journals Efek vitamin D terhadap kadar IL-10, IFN-γ, dan histamin pada kultur PBMC pasien rinitis alergi

2018 ◽  
Vol 47 (2) ◽  
pp. 132
Author(s):  
Budi Utama ◽  
Heri Wibowo ◽  
Niken Lestari Poerbonegoro
Keyword(s):  
Vitamin D ◽  
Allergic Rhinitis ◽  
Peripheral Blood Mononuclear Cells ◽  
House Dust ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Ifn Γ

Latar belakang: Rinitis alergi (RA) adalah penyakit inflamasi pada hidung, yang disebabkan oleh reaksi alergi pada pasien atopi. Tungau debu rumah merupakan aeroalergen yang tersering memicu reaksi alergi. Pada tahun 1988, reseptor vitamin D berhasil dilakukan klon. Reseptor vitamin D berlokasi di beberapa jaringan dan sel tubuh manusia, termasuk di sel-sel darah tepi berinti tunggal (peripheral blood mononuclear cells/PMBC). Tujuan: Mengidentifikasi pengaruh pemberian vitamin D pada sel-sel darah tepi berinti tunggal penderita rinitis alergi terhadap sel Th1, Th2, dan T Regulator, dengan cara melihat sekresi IFN-γ, IL-10, dan histamin. Metode: Sampel berupa darah segar (whole blood) penderita rinitis alergi yang telah dilakukan prick test, diolah dengan metode Ficoll untuk mengisolasi sel berinti tunggal. Kultur sel limfosit sebelum perlakuan dibagi menjadi kelompok yang diberi pendedahan dengan 1,25(OH)2D3 100 nM dan tanpa pendedahan, waktu inkubasi 7 hari, dengan penambahan phytohaemaglutinin dan alergen tungau pada hari ke-4. Kultur sel-sel darah tepi berinti tunggal dari pasien RA setelah perlakuan, selanjutnya pada hari ke-7 supernatannya diambil dan dibagi untuk diukur kadar sitokin IFN-γ, IL-10, dan histamin secara ELISA. Dilakukan uji secara statistik untuk melihat pola dari tiap parameter. Hasil: Pemberian alergen tungau tanpa vitamin D menyebabkan meningkatnya kadar histamin serta menurunkan kadar IL-10 dan IFN-γ. Pemberian vitamin D pada kultur sel darah tepi berinti tunggal yang telah diberi alergen tungau, dapat meningkatkan kadar IL-10 dan menurunkan kadar IFN-γ, serta histamin. Kesimpulan: Menurunnya kadar histamin dan IFN-γ terhadap stimulasi alergen tungau pada pasien rinitis alergi yang diberi vitamin D cenderung berhubungan dengan meningkatnya kadar IL-10. Kata kunci: Sel mast, rinitis alergi, tungau debu rumah, vitamin D ABSTRACT Background: Allergic rhinitis is an inflammatory disease of the nose, caused by an allergic reaction in atopic patients. House dust mites are the most common aeroalergen. In 1988, vitamin D receptor had been cloned successfully. Vitamin D receptors are located in various tissues and human body cells, including peripheral blood mononuclear cells (PBMCs). Purpose: To identify the effect of vitamin D on peripheral blood mononuclear cells culture of allergic rhinitis patients towards Th1, Th2, and T Regulator cell, by identifying IL-10, IFN-γ, and histamine secretion levels. Method: The sample were obtained from fresh blood (whole blood) of allergic rhinitis patients who had been prick tested, and isolated by Ficoll method. Pre-treated lymphocyte culture divided into groups treated with and without 1,25(OH)2D3 100 nM, and incubated for 7 days, with addition of phytohaemaglutinin and allergen mites on day 4. Cultures of PBMC cells after treatment were harvested on day 7, then the supernatant was dialyzed to measure the levels of IFN-γ IL-10 and histamine cytokines. Statistical test was performed to identify patterns of each parameter. Results: Treatment of allergen mites without vitamin D could increase levels of histamine and lower levels of IL-10 and IFN-γ. Provision of vitamin D in PBMC cell culture that had been given allergen mites could increase levels of IL-10 and decreased levels of IFN-γ and histamine. Conclusion: Lower levels of histamine and IFN-γ against allergen mite stimulation of allergic rhinitis patients who were given vitamin D tend to be associated with increased IL-10 levels. Keywords: Mast cell, allergic rhinitis, house dust mite, vitamin D

scholarly journals Effect of Steroids on Lipopolysaccharide/Interleukin 2-Induced Interleukin 18 Production in Peripheral Blood Mononuclear Cells

2002 ◽  
Vol 30 (2) ◽  
pp. 144-160 ◽  
Author(s):  
M Kodama ◽  
HK Takahashi ◽  
H Iwagaki ◽  
H Itoh ◽  
T Morichika ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Steroid Treatment ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Enhanced Production ◽  
Blood Mononuclear Cells ◽  
Ifn Γ ◽  
Cytokine Cascade

Interleukin (IL) 18, a powerful inducer of the immunoregulatory cytokine interferon-γ (IFN-γ), presents upstream of the cytokine activation cascade in the inflammatory response. The anti-inflammatory properties of steroids permit their use in various conditions, although effects are transient and pathological states are not fully relieved by short-term steroidal use. We examined the effect of lipopolysaccharide (LPS)/IL-2 on the cytokine cascade in human peripheral blood mononuclear cells (PBMCs). We also examined the effect of steroids on LPS/IL-2-induced cytokine production in human PBMCs taken from healthy volunteers. Cell-free supernatant fractions were assayed for IL-18, IL-12, IL-2, IFN-γ and IL-10 protein, using enzyme-linked immunosorbent assays, and synergy between LPS and IL-2 in enhanced production of IL-18 was observed. Steroids suppressed the production of IL-18 and other secondary cytokines in LPS/IL-2-stimulated PBMCs, in a concentration- and time-dependent manner, although inhibition was incomplete even at high concentrations. Effects of steroid treatment on expression of membrane-bound LPS receptor antigen (mCD14) and intercellular adhesion molecule-1 (ICAM-1) in PBMCs were studied by flow cytometric analysis. Steroid treatment up-regulated mCD14 expression in a concentration-dependent manner, with no effect on ICAM-1 expression. These results suggest that the incomplete counteraction of steroids in the LPS/IL-2-initiating cytokine cascade is due, at least partly, to the up-regulation of mCD14 by steroid preparations, which increases susceptibility to bacterial endotoxins.

scholarly journals Monitoring Tacrolimus Concentrations in Whole Blood and Peripheral Blood Mononuclear Cells: Inter- and Intra-Patient Variability in a Cohort of Pediatric Patients

2021 ◽  
Vol 12 ◽  
Author(s):  
Amedeo De Nicolò ◽  
Michele Pinon ◽  
Alice Palermiti ◽  
Antonello Nonnato ◽  
Alessandra Manca ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Pediatric Patients ◽  
Mononuclear Cells ◽  
Individual Variability ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Intracellular Concentrations ◽  
The One

Tacrolimus (TAC) is a first-choice immunosuppressant for solid organ transplantation, characterized by high potential for drug-drug interactions, significant inter- and intra-patient variability, and narrow therapeutic index. Therapeutic drug monitoring (TDM) of TAC concentrations in whole blood (WB) is capable of reducing the incidence of adverse events. Since TAC acts within lymphocytes, its monitoring in peripheral blood mononuclear cells (PBMC) may represent a valid future alternative for TDM. Nevertheless, TAC intracellular concentrations and their variability are poorly described, particularly in the pediatric context. Therefore, our aim was describing TAC concentrations in WB and PBMC and their variability in a cohort of pediatric patients undergoing constant immunosuppressive maintenance therapy, after liver transplantation. TAC intra-PBMCs quantification was performed through a validated UHPLC–MS/MS assay over a period of 2–3 months. There were 27 patients included in this study. No significant TAC changes in intracellular concentrations were observed (p = 0.710), with a median percent change of −0.1% (IQR −22.4%–+46.9%) between timings: this intra-individual variability was similar to the one in WB, −2.9% (IQR −29.4–+42.1; p = 0.902). Among different patients, TAC weight-adjusted dose and age appeared to be significant predictors of TAC concentrations in WB and PBMC. Intra-individual seasonal variation of TAC concentrations in WB, but not in PBMC, have been observed. These data show that the intra-individual variability in TAC intracellular exposure is comparable to the one observed in WB. This opens the way for further studies aiming at the identification of therapeutic ranges for TAC intra-PBMC concentrations.

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