The aim of this study was to provide proof-of-concept for quantitative and qualitative label-free detection of immune complexes through myeloid cells with imaging surface plasmon resonance. Surface plasmon resonance imaging was first applied to monitor the binding of human sera from healthy and RA patients to immobilised citrullinated RA specific peptide antigens, HCP2 and VCP2. Next, the binding of monocytoid cell line U937 to the resulting immune complexes on the sensor surface was monitored. As control, binding of U937 was monitored to IgG subclasses simultaneously. Cell response results were compared to results of CCP2 ELISA, clinical RA diagnosis and antigen specific antibody distribution of the samples. Human IgG3 triggered the most pronounced response followed by IgG1 and IgG4, while IgG2 did not result in U937 cell binding. Serum samples obtained from RA patients resulted in significantly increased cell response to VCP2 compared to healthy controls. The strength of cell response towards VCP2-immune complexes showed significant correlation with levels of antigen specific IgA, IgG and IgG3. Cellular responses on VCP2 immune complexes showed significant association with both CCP2-based serological positivity and EULAR criteria-based clinical RA diagnosis. Immunoglobulin-triggered binding of monocytoid cells can be monitored using a label-free multiplex technology. Since these binding events are presumably initiated by Fc receptors, the system provides a tool for biological detection of autoantibodies with diagnostic value, here exemplified by anti-citrullinated antibodies. This provides added information to antibody levels, as interaction with Fc-receptor-expressing cells are also affected by post-translational modification of the immunoglobulins. This article is protected by copyright. All rights reserved.
Label-free detection of immune complexes with myeloid cells / Szittner, Zoltán; Bentlage, Arthur E H; Rovero, Paolo; Migliorini, Paola; Lóránd, Veronika; Prechl, József; Vidarsson, Gestur. - In: CLINICAL AND EXPERIMENTAL IMMUNOLOGY. - ISSN 0009-9104. - ELETTRONICO. - 185:(2016), pp. 72-80. [10.1111/cei.12788]
Label-free detection of immune complexes with myeloid cells
ROVERO, PAOLO;
2016
Abstract
The aim of this study was to provide proof-of-concept for quantitative and qualitative label-free detection of immune complexes through myeloid cells with imaging surface plasmon resonance. Surface plasmon resonance imaging was first applied to monitor the binding of human sera from healthy and RA patients to immobilised citrullinated RA specific peptide antigens, HCP2 and VCP2. Next, the binding of monocytoid cell line U937 to the resulting immune complexes on the sensor surface was monitored. As control, binding of U937 was monitored to IgG subclasses simultaneously. Cell response results were compared to results of CCP2 ELISA, clinical RA diagnosis and antigen specific antibody distribution of the samples. Human IgG3 triggered the most pronounced response followed by IgG1 and IgG4, while IgG2 did not result in U937 cell binding. Serum samples obtained from RA patients resulted in significantly increased cell response to VCP2 compared to healthy controls. The strength of cell response towards VCP2-immune complexes showed significant correlation with levels of antigen specific IgA, IgG and IgG3. Cellular responses on VCP2 immune complexes showed significant association with both CCP2-based serological positivity and EULAR criteria-based clinical RA diagnosis. Immunoglobulin-triggered binding of monocytoid cells can be monitored using a label-free multiplex technology. Since these binding events are presumably initiated by Fc receptors, the system provides a tool for biological detection of autoantibodies with diagnostic value, here exemplified by anti-citrullinated antibodies. This provides added information to antibody levels, as interaction with Fc-receptor-expressing cells are also affected by post-translational modification of the immunoglobulins. This article is protected by copyright. All rights reserved.File | Dimensione | Formato | |
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