| 1. |
- Aljadi, Zenib, et al.
(författare)
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A novel tool for clinical diagnosis of allergy operating a microfluidic immunoaffinity basophil activation test technique
- 2019
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Ingår i: Clinical Immunology. - : ACADEMIC PRESS INC ELSEVIER SCIENCE. - 1521-6616 .- 1521-7035. ; 209
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Tidskriftsartikel (refereegranskat)abstract
- The Basophil Activation Test (BAT) is a valuable allergy diagnostic tool but is time-consuming and requires skilled personnel and cumbersome processing, which has limited its clinical use. We therefore investigated if a microfluidic immunoaffinity BAT (miBAT) technique can be a reliable diagnostic method. Blood was collected from allergic patients and healthy controls. Basophils were challenged with negative control, positive control (anti-FccRI), and two concentrations of a relevant and non-relevant allergen. CD203c and CD63 expression was detected by fluorescent microscopy and flow cytometry. In basophils from allergic patients the CD63% was significantly higher after allergen activation as compared to the negative control (p < .0001-p = .0004). Activation with non-relevant allergen showed equivalent CD63% expression as the negative control. Further, the miBAT data were comparable to flow cytometry. Our results demonstrate the capacity of the miBAT technology to measure different degrees of basophil allergen activation by quantifying the CD63% expression on captured basophils.
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| 2. |
- Aljadi, Zenib, et al.
(författare)
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Altered basophil function in patients with chronic kidney disease on hemodialysis
- 2017
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Ingår i: Clinical Nephrology. - : DUSTRI-VERLAG DR KARL FEISTLE. - 0301-0430. ; 88:2, s. 86-96
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Tidskriftsartikel (refereegranskat)abstract
- Aims: Chronic kidney disease (CKD) leads to impairment of immune cell function. Given the potential role of basophils in the pathogenesis of CKD, we aimed to study the basophil responsiveness towards microbial antigen exposure, judged as adhesion molecule expression and degranulation, in CKD patients on hemodialysis. Materials and methods: We selected markers linked to two crucial biological phases: the transmigration and degranulation processes, respectively. For the transmigration process, we selected the adhesion molecules CD11b, active CD11b epitope, and CD62L and for the degranulation process CD203c (piecemeal degranulation marker), CD63 (degranulation marker), and CD300a (inhibitory marker of degranulation). We measured basophil responsiveness after stimulation of different activation pathways in basophils using lipopolysaccharide (LPS), peptidoglycan (PGN), formyl-methyinoyl-leucyl-phenylalanine (fMLP), and anti-FceRI-ab. Results: The expression of CD63 in basophils following activation by fMLP was significantly higher in the patient group compared to matched healthy controls, but no differences were observed after activation by anti-Fc.RI. CD300a expression was significantly higher in patients following activation by fMLP and anti-Fc.RI, and the active epitope CD11b expression was significantly higher in patients after LPS activation. In addition, we found that CD62L was not shed from the cell surface after activation with LPS and fMLP. A slight downregulation was noted after activation with anti-Fc.RI in healthy controls. Conclusion: Together, these data demonstrate that basophil functions related to adhesion and degranulation are altered in CKD patients on hemodialysis, which indicates a potential role for the basophil in the pathogenesis of complications related to infections.
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| 3. |
- Aljadi, Zenib, et al.
(författare)
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Microfluidic Immunoaffinity Basophil Activation Test for Point-of-Care Allergy Diagnosis
- 2019
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Ingår i: Journal of Applied Laboratory Medicine (JALM). - : American Association for Clinical Chemistry. - 2475-7241 .- 2576-9456. ; 4:2, s. 152-163
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Tidskriftsartikel (refereegranskat)abstract
- Background: The flow cytometry-based basophil activation test (BAT) is used for the diagnosis of allergic response. However, flow cytometry is time-consuming, requiring skilled personnel and cumbersome processing, which has limited its use in the clinic. Here, we introduce a novel microfluidic-based immunoaffinity BAT (miBAT) method. Methods: The microfluidic device, coated with anti-CD203c, was designed to capture basophils directly from whole blood. The captured basophils are activated by anti-FceRI antibody followed by optical detection of CD63 expression (degranulation marker). The device was first characterized using a basophil cell line followed by whole blood experiments. Weevaluated the device with ex vivo stimulation of basophils in whole blood from healthy controls and patients with allergies and compared it with flow cytometry. Results: The microfluidic device was capable of capturing basophils directly from whole blood followed by in vitro activation and quantification of CD63 expression. CD63 expression was significantly higher (P = 0.0002) in on-chip activated basophils compared with nonactivated cells. The difference in CD63 expression on anti-FceRI-activated captured basophils in microfluidic chip was significantly higher (P = 0.03) in patients with allergies compared with healthy controls, and the results were comparable with flow cytometry analysis (P = 0.04). Furthermore, there was no significant difference of CD63% expression in anti-FceRI-activated captured basophils in microfluidic chip compared with flow cytometry. Conclusions: We report on the miBAT. This device is capable of isolating basophils directly from whole blood for on-chip activation and detection. The new miBAT method awaits validation in larger patient populations to assess performance in diagnosis and monitoring of patients with allergies at the point of care.
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| 4. |
- Rikard, S. M., et al.
(författare)
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Multiple computational modeling approaches for prediction of wound healing dynamics following pharmacologic intervention
- 2017
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Ingår i: Biomedical Engineering Society (BMES) annual meeting, Phoenix, AZ, USA, 11-14 October 2017.
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Konferensbidrag (refereegranskat)abstract
- Diabetic wounds are known to have a delayed course of wound healing. We have recently demonstrated that injection of a synthetic modified RNA (modRNA) that enhances VEGF-A protein expression accelerates healing of full-thickness cutaneous wounds in db/db diabetic mice. Here, we compare two different computational modeling approaches to explore how the dosing amount and time course affect the rate of wound healing. We show that a partial differential equation (PDE) model is appropriate for questions concerning spatial resolution of healing throughout the wound, while a nonlinear mixed effect model (NLME) is more appropriate for capturing population level variations in healing rate when dealing with a sparse data set. Both models display sensitivity to varying dosing amount and timing.
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