Bettina Engel1, Gregor Müller2, Beate Roch3, Hans-Egbert Schröder3, Martin Aringer3, Stefan R Bornstein4, Henning Morawietz5. 1. Division of Vascular Endothelium and Microcirculation, Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; Institut für Hausarztmedizin, Medizinische Fakultät Bonn, Bonn, Germany. 2. Division of Vascular Endothelium and Microcirculation, Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany. 3. Division of Rheumatology, Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany. 4. Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany. 5. Division of Vascular Endothelium and Microcirculation, Department of Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany. Electronic address: Henning.Morawietz@tu-dresden.de.
Abstract
OBJECTIVE: The antiphospholipid syndrome (APS) is a systemic auto-immune disease with an unclear pathophysiology. The aim of our study was to understand the development of APS on a cellular level. Therefore, we analyzed the influence of human serum of APS patients on endothelial expression of specific genes and proteins in comparison to a control group. METHODS: In this study, we analyzed the expression of ICAM-1, VCAM-1, E-selectin and annexin V in primary cultures of human umbilical vein endothelial cells (HUVEC) in response to 10% (v/v) serum of control patients (n = 6), patients with systemic lupus erythematosus (SLE) and no APS (n = 4) or APS patients (n = 9) for 24 h. Total RNA was prepared from confluent endothelial cell layers and mRNA expression of ICAM-1, VCAM-1 and E-selectin was analyzed by reverse transcription polymerase-chain reaction (RT-PCR). The protein expression was determined by Western blot. Serum protein concentrations of soluble forms of adhesion molecules sICAM-1 and sVCAM-1 were quantified by ELISA. Gene expression data were correlated with clinical parameters. RESULTS: The mRNA expression of ICAM-1 was increased in cells incubated with serum from APS patients (166 ± 22% of control; P = 0.023). Serum of patients with (SLE)/no APS caused a 1.4-fold higher ICAM-1 mRNA level. Western blot analysis showed an increase in protein expression of adhesion molecules ICAM-1 (260 ± 49%; P = 0.011) and VCAM-1 (357 ± 97%; P = 0.023) in cells that were incubated with serum from APS patients. Plasma analysis showed elevated levels of sVCAM-1 in APS patients (189 ± 34%; P = 0.045) compared to the levels measured in the control group. The sVCAM-1 plasma level was correlating with the frequency of abortions. CONCLUSION: An augmented expression of endothelial adhesion molecules is involved in the pathophysiology of patients with antiphospholipid syndrome.
OBJECTIVE: The antiphospholipid syndrome (APS) is a systemic auto-immune disease with an unclear pathophysiology. The aim of our study was to understand the development of APS on a cellular level. Therefore, we analyzed the influence of human serum of APSpatients on endothelial expression of specific genes and proteins in comparison to a control group. METHODS: In this study, we analyzed the expression of ICAM-1, VCAM-1, E-selectin and annexin V in primary cultures of human umbilical vein endothelial cells (HUVEC) in response to 10% (v/v) serum of control patients (n = 6), patients with systemic lupus erythematosus (SLE) and no APS (n = 4) or APSpatients (n = 9) for 24 h. Total RNA was prepared from confluent endothelial cell layers and mRNA expression of ICAM-1, VCAM-1 and E-selectin was analyzed by reverse transcription polymerase-chain reaction (RT-PCR). The protein expression was determined by Western blot. Serum protein concentrations of soluble forms of adhesion molecules sICAM-1 and sVCAM-1 were quantified by ELISA. Gene expression data were correlated with clinical parameters. RESULTS: The mRNA expression of ICAM-1 was increased in cells incubated with serum from APSpatients (166 ± 22% of control; P = 0.023). Serum of patients with (SLE)/no APS caused a 1.4-fold higher ICAM-1 mRNA level. Western blot analysis showed an increase in protein expression of adhesion molecules ICAM-1 (260 ± 49%; P = 0.011) and VCAM-1 (357 ± 97%; P = 0.023) in cells that were incubated with serum from APSpatients. Plasma analysis showed elevated levels of sVCAM-1 in APSpatients (189 ± 34%; P = 0.045) compared to the levels measured in the control group. The sVCAM-1 plasma level was correlating with the frequency of abortions. CONCLUSION: An augmented expression of endothelial adhesion molecules is involved in the pathophysiology of patients with antiphospholipid syndrome.
Authors: Laura Whittall-Garcia; Kirubel Goliad; Michael Kim; Dennisse Bonilla; Dafna Gladman; Murray Urowitz; Paul R Fortin; Eshetu G Atenafu; Zahi Touma; Joan Wither Journal: Front Immunol Date: 2022-05-30 Impact factor: 8.786
Authors: Manuela Velásquez; Luisa F Peláez; Mauricio Rojas; Raúl Narváez-Sánchez; Jesús A Velásquez; Carlos Escudero; Sebastián San Martín; Ángela P Cadavid Journal: Front Physiol Date: 2021-12-02 Impact factor: 4.566