Development of a single-antigen magnetic bead assay (SAMBA) for the sensitive detection of HPA-1a alloantibodies using tag-engineered recombinant soluble β3 integrin

Skaik, Younis; Battermann, Anja; Hiller, Oliver; Meyer, Oliver; Figueiredo, Constança; Salama, Abdulgabar; Blasczyk, Rainer

doi:10.1016/j.jim.2013.02.011

Cited by 5 publications

(9 citation statements)

References 59 publications

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“…Previously, we have established an ELISA for the detection of sβ 3– integrin [19]. In this study, we have detected sβ 3– integrin in 35% of AML patients (n = 8) prior chemotherapy.…”

Section: Resultsmentioning

confidence: 89%

“…Therefore, it is important to evaluate the effect of proteins specifically detected in leukaemia patients in NK cell function. In this study, all functional assays were performed using recombinant sβ3 - integrin which was produced as previously described [19]. Fluorescence microscopy analyses showed that sβ 3 -integrin is capable of binding NK cells (Figure 3A).…”

Section: Resultsmentioning

confidence: 99%

“…Soluble β 3- integrin was produced recombinantly in HEK293 as previously described [19]. The recombinant sβ 3- integrin sequence was designed based on the RNA sequence detected in the AML patients showing increased serum levels of sβ 3- integrin.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, a lentiviral vector encoding the sequence for sβ 3- integrin was used for the HEK293 cells transduction in the presence of 8 µg/mL protamine sulphate (Sigma Aldrich Chemie, Munich, Germany). Seventy-two hours after transduction, cell culture supernatant was analyzed by ELISA for the presence of sβ 3- integrin as previously described [19]. sβ 3- integrin producing HEK 293 cells were cultured in bioreactors ( CELLine Adherent, Integra, Fernwald, Germany) to increase the protein production yields.…”

Section: Methodsmentioning

confidence: 99%

“…Known amounts of V5-His-tagged β 3- integrin protein were used as the reference standard. Produced sβ 3- integrin was controlled for quality controlled by sodium dodecylsulfate (SDS) and immunoblot as previously described [19].…”

Section: Methodsmentioning

confidence: 99%

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Secreted β3-Integrin Enhances Natural Killer Cell Activity against Acute Myeloid Leukemia Cells

et al. 2014

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Integrins are a large family of heterodimeric proteins that are involved in cell adhesion, migration, and proliferation. Integrin diversity and function is regulated by alternative splicing. Membrane-bound and truncated β3-integrins were shown to be key players in cancer metastasis. However, the immunomodulatory functions of the soluble (s) β3-integrin have not been investigated yet. In this study, we described a novel form of sβ3-integrin in acute myeloid leukaemia (AML) patients. Furthermore, we assessed the role of the sβ3-integrin in the modulation of natural killer (NK)-cell activity. Levels of sβ3-integrin were analysed in plasma samples of 23 AML patients and 26 healthy donors by ELISA. The capacity of sβ3-integrin to regulate NK cell activity was investigated using proliferation, cytokine secretion, and cytotoxicity assays. Circulating sβ3-integrin was detected in the plasma of 8 AML patients. NK cells showed significantly higher proliferation rates after stimulation with sβ3-integrin and IL-2, IL-15 (73%). Significant increases in the NK cells’ secreted levels of TNF-α, IFN-γ were measured in presence of sβ3-integrin. In addition, sβ3-integrin caused the upregulation of Granzyme B transcripts levels as well as FasL expression levels in NK cells. Most importantly, significantly higher K562 or AML blast target cell lysis rates were observed when NK cells were exposed to sβ3-integrin. This study reports the identification of a novel sβ3-integrin in AML patients and provides novel insights into its role in the immunomodulation of NK cell activity.

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“…Previously, we have established an ELISA for the detection of sβ 3– integrin [19]. In this study, we have detected sβ 3– integrin in 35% of AML patients (n = 8) prior chemotherapy.…”

Section: Resultsmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Secreted β3-Integrin Enhances Natural Killer Cell Activity against Acute Myeloid Leukemia Cells

et al. 2014

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Progress and development of platelet antibody detection

Porcelijn

Huiskes

Haas

2020

Transfusion and Apheresis Science

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Human platelet antibody (HPA) detection is necessary for the diagnosis and therapeutic decisions for refractoriness to platelet transfusions, post transfusion purpura and fetal and neonatal alloimmune thrombocytopenia. In the last four to five decades many new developments, both in knowledge and methods, have increased the quality of platelet serology. However, the quest for the optimal antibody detection method(s) encountered, sometimes unexpected, difficulties. In this review the various aspects concerning platelet antibody test methods and detection of platelet antibodies both for the diagnostic and screening setting are discussed.

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Establishment of a Multivariate Analysis Based on Photonic Crystal Hydrogel Bead Arrays and Its Application for Detecting Platelet-specific Antibodies

et al. 2018

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Development of a single-antigen magnetic bead assay (SAMBA) for the sensitive detection of HPA-1a alloantibodies using tag-engineered recombinant soluble β3 integrin

Cited by 5 publications

References 59 publications

Secreted β3-Integrin Enhances Natural Killer Cell Activity against Acute Myeloid Leukemia Cells

Secreted β3-Integrin Enhances Natural Killer Cell Activity against Acute Myeloid Leukemia Cells

Progress and development of platelet antibody detection

Establishment of a Multivariate Analysis Based on Photonic Crystal Hydrogel Bead Arrays and Its Application for Detecting Platelet-specific Antibodies

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