Detection of Autoantibodies Against the β1-Adrenergic Receptor in the Sera of Patients via the Competitive cell-Based Enzyme Linked Immunosorbent Assay

Shevelev, Alexander; Kostiukevich, M V; Ефремов, Е. Е.; Власик, Т. Н.; Миронова, Н. А.; Zykov, K. A.; Kashirina, Natalia M.; Kuznetsova, I B; Шарф, Т. В.; Mamochkina, E N; Липатова, Л.Н.; Peklo, M. M.; Rutkevich, P. N.; Yanushevskaya, E. V.; Rybalkin, I. N.; Стукалова, О. В.; Малкина, Т. А.; Belyaeva, M M; Kuznetsova, T V; Tkachev, G A; Zinchenko, L V; Gupalo, Elena; Agapova, Olga; Yureneva-Tkhorzhevskaya, T V; Rvacheva, A. V.; Sidorova, M. V.; Sadgyan, A S; Tereshchenko, S. N.; Golitsyn, S. P.

doi:10.18565/cardio.2016.11.61-70

Cited by 5 publications

(6 citation statements)

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“…В качестве продуцента АДРБ1 в функционально активном состоянии использовали линию клеток ADL-7A, полученную нами ранее путем трансфекции клеток HEK293 плазмидой, содержащей ген АДРБ1, с последующим отбором стабильных трансфектантов. Получение, характеристика и условия культивирования описаны в работе [23].…”

Section: методикаunclassified

β1-adrenergic Receptor Solubilized in the Form of Nanodiscs: Screening of Various Amphipatic Polymers

Peclo,

Lipatova,

Kashirina

et al. 2023

BMCRM

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The development of a reliable and easily used diagnostic test for measuring autoantibodies to ?1-adrenergic receptor (?1ADR Ab) in patient blood is an unmet clinical need. The enzyme-linked immunosorbent assay (ELISA) is considered as the most appropriate method for this task. In ELISA, the use of peptides corresponding to various fragments of amino acid sequence of ?1ADR as antigens leads to inadequate results as β1ADR Ab appear to recognize conformationally dependent epitopes that are generated during the formation of unique tertiary structure of the receptor. Isolation of ?1ADR preserving the native conformation and functional characteristics is a quite challenging task. A promising approach to address this task is the use of amphipatic polymers capable of forming nanodiscs, it permits to successfully solubilize membrane proteins. In order to obtain the preparations of solubilized β1ADR that can be used as antigens in ELISA we have tested 17 various amphipatic polymers. The best relative solubilization values (RSV) were obtained using UltrasoluteTM Amphipol 17 (87%) and 18 (62%), as well as by AASTY 11-45 (76%), 11-50 (77%) and 6-50 (78.5%).

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Section: методикаunclassified

β1-adrenergic Receptor Solubilized in the Form of Nanodiscs: Screening of Various Amphipatic Polymers

Peclo,

Lipatova,

Kashirina

et al. 2023

BMCRM

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“…Subsequently, other researchers found that about 40%‐60% of patients with cardiac dysfunction were detected with β 1 ‐AA in serum 4 . It can bind with β 1 ‐adrenoceptor (β 1 ‐AR) on the surface of cardiomyocyte membrane to cause persistent injury of cardiomyocytes, induce cardiomyocyte death and eventually lead to cardiac dysfunction 4 . Our team originally found that β 1 ‐AA could induce the decrease of myocardial autophagic flux, which contributed to cardiac dysfunction 5 .…”

Section: Introductionmentioning

confidence: 95%

“…In 1987, the researcher first discovered β 1 ‐adrenergic receptor autoantibody (β 1 ‐AA) as a product of immune disorder in serum of patients with dilated cardiomyopathy 3 . Subsequently, other researchers found that about 40%‐60% of patients with cardiac dysfunction were detected with β 1 ‐AA in serum 4 . It can bind with β 1 ‐adrenoceptor (β 1 ‐AR) on the surface of cardiomyocyte membrane to cause persistent injury of cardiomyocytes, induce cardiomyocyte death and eventually lead to cardiac dysfunction 4 .…”

Section: Introductionmentioning

confidence: 99%

Adiponectin up‐regulates the decrease of myocardial autophagic flux induced by β₁‐adrenergic receptor autoantibody partly dependent on AMPK

Sun

Long

et al. 2021

J Cellular Molecular Medi

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Cardiomyocytes autophagy is essential for maintaining cardiac function. Our previous studies have found that β1‐adrenergic receptor autoantibody (β1‐AA) induced the decreased myocardial autophagic flux, which resulted in cardiomyocyte death and cardiac dysfunction. And other studies demonstrated that β1‐AA induced the decrease of AMPK phosphorylation, the key hub of autophagy pathway, while adiponectin up‐regulated autophagic flux mediated by AMPK. However, it is not clear whether adiponectin improves the inhibition of myocardial autophagic flux induced by β1‐AA by up‐regulating the level of AMPK phosphorylation. In this study, it has been confirmed that β1‐AA induced the decrease of AMPK phosphorylation level in both vivo and vitro. Moreover, pretreatment of cardiomyocytes with AMPK inhibitor Compound C could further reduce the autophagic flux induced by β1‐AA. Adiponectin deficiency could aggravate the decrease of myocardial AMPK phosphorylation level, autophagic flux and cardiac function induced by β1‐AA. Further, exogenous adiponectin could reverse the decline of AMPK phosphorylation level and autophagic flux induced by β1‐AA and even reduce cardiomyocyte death. While pretreated with the Compound C, the adiponectin treatment did not improve the decreased autophagosome formation, but still improved the decreased autophagosome clearance induced by β1‐AA in cardiomyocytes. This study is the first time to confirm that β1‐AA could inhibit myocardial autophagic flux by down‐regulating AMPK phosphorylation level. Adiponectin could improve the inhibition of myocardial autophagic flux induced by β1‐AA partly dependent on AMPK, so as to provide an experimental basis for the treatment of patients with β1‐AA‐positive cardiac dysfunction.

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“…In 1987, the researcher rst discovered β 1 -adrenergic receptor antibody (β 1 -AA) as a product of immune disorder in serum of patients with dilated cardiomyopathy [3] . Subsequently, other researchers found that about 40% − 60% of patients with cardiac dysfunction were detected with β 1 -AA in serum [4] . It can bind with β 1 -adrenoceptor (β 1 -AR) on the surface of cardiomyocytes membrane to cause persistent injury of cardiomyocytes, induce cardiomyocyte death and eventually lead to cardiac dysfunction [4] .…”

Section: Introductionmentioning

confidence: 97%

“…Subsequently, other researchers found that about 40% − 60% of patients with cardiac dysfunction were detected with β 1 -AA in serum [4] . It can bind with β 1 -adrenoceptor (β 1 -AR) on the surface of cardiomyocytes membrane to cause persistent injury of cardiomyocytes, induce cardiomyocyte death and eventually lead to cardiac dysfunction [4] . Our team originally found that β 1 -AA could induce the decrease of myocardial autophagic ux, which contributed to cardiac dysfunction [5] .…”

Section: Introductionmentioning

confidence: 97%

Adiponectin up-regulates the decrease of myocardial autophagic flux induced by β1-adrenergic receptor antibody partly dependent on AMPK

Sun

Long

et al. 2021

Preprint

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BackgroundCardiomyocyte autophagy is essential for maintaining cardiac function. Our previous studies have found that β1-adrenergic receptor autoantibody (β1-AA) induced the decreased myocardial autophagic flux, which resulted in cardiomyocytes death and cardiac dysfunction. And other studies demonstrated that β1-AA induced the decrease of AMPK phosphorylation, the key hub of autophagy pathway, while adiponectin up-regulated autophagic flux mediated by AMPK. However, it is not clear whether adiponectin improves the inhibition of myocardial autophagic flux induced by β1-AA by up-regulating the level of AMPK phosphorylation.MethodAPN-KO and WT mice were subcutaneously injected withthe peptide of the second extracellular loop of β1-adrenergic receptor (β1-AR-ECII) as antigen to establish β1-AA active immunization model. The OD value of β1-AA in serum of mice was detected by SA-ELISA to ensure the successful construction of active immunization model. The changes of cardiac function and myocardial fibrosis were detected by small animal ultrasound and Masson trichrome staining. The cardiomyocytes viability was assessed using a Cell Counting Kit-8. The mRNA levels of LC3B and Beclin1 in cardiomyocytes were measured by Real-time PCR. The protein levels of LC3 II, p62, AMP dependent protein kinase (AMP) - activated protein kinase (AMPK) and phosphorylated AMP dependent protein kinase (p-AMPK) in cardiomyocytes were detected by Western blotting.ResultsIn this study, it has been confirmed that β1-AA induced the decrease of AMPK phosphorylation level in both vivo and vitro. Moreover, pretreatment of cardiomyocytes with AMPK inhibitor Compound C could further reduce the autophagic flux induced by β1-AA. Adiponectin deficiency could aggravate the decrease of myocardial AMPK phosphorylation level, autophagic flux and cardiac function induced by β1-AA. Further, exogenous adiponectin could reverse the decline of AMPK phosphorylation level and autophagic flux induced by β1-AA, and even reduce cardiomyocytes death. While pre-treated with the Compound C, the adiponectin treatment did not improve the decreased autophagosome formation, but still improved the decreased autophagosome clearance induced by β1-AA in cardiomyocytes, suggesting that adiponectin up-regulated β1-AA-induced decreased autophagic flux of cardiomyocytes partly depended on the AMPK.ConclusionThis study is the first time to confirm that β1-AA could inhibit myocardial autophagic flux by down regulating AMPK phosphorylation level. Adiponectin could improve the inhibition of myocardial autophagic flux induced by β1-AA partly dependent on AMPK, so as to provide an experimental basis for the treatment of patients with β1-AA-positive cardiac dysfunction.

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Detection of Autoantibodies Against the β1-Adrenergic Receptor in the Sera of Patients via the Competitive cell-Based Enzyme Linked Immunosorbent Assay

Cited by 5 publications

References 0 publications

β1-adrenergic Receptor Solubilized in the Form of Nanodiscs: Screening of Various Amphipatic Polymers

β1-adrenergic Receptor Solubilized in the Form of Nanodiscs: Screening of Various Amphipatic Polymers

Adiponectin up‐regulates the decrease of myocardial autophagic flux induced by β₁‐adrenergic receptor autoantibody partly dependent on AMPK

Adiponectin up-regulates the decrease of myocardial autophagic flux induced by β1-adrenergic receptor antibody partly dependent on AMPK

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Detection of Autoantibodies Against the β1-Adrenergic Receptor in the Sera of Patients via the Competitive cell-Based Enzyme Linked Immunosorbent Assay

Cited by 5 publications

References 0 publications

β1-adrenergic Receptor Solubilized in the Form of Nanodiscs: Screening of Various Amphipatic Polymers

β1-adrenergic Receptor Solubilized in the Form of Nanodiscs: Screening of Various Amphipatic Polymers

Adiponectin up‐regulates the decrease of myocardial autophagic flux induced by β1‐adrenergic receptor autoantibody partly dependent on AMPK

Adiponectin up-regulates the decrease of myocardial autophagic flux induced by β1-adrenergic receptor antibody partly dependent on AMPK

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Adiponectin up‐regulates the decrease of myocardial autophagic flux induced by β₁‐adrenergic receptor autoantibody partly dependent on AMPK