A novel assay for detecting virus-specific antibodies triggering activation of Fc¦Ã receptors

详细信息    查看全文

• 作者：Eugenia Corrales-Aguilar^a ; ¹ ; Mirko Trilling^a ; ² ; Henrike Reinhard^a ; Eva Mercé ; -Maldonado^a ; Marek Widera^a ; Heiner Schaal^a ; Albert Zimmermann^a ; Ofer Mandelboim^b ; Hartmut Hengel^a ; ^{Hartmut.Hengel@uni-duesseldorf.de}
• 关键词：ADCC ; antibody-dependent cellular cytotoxicity ; EBV ; Epstein&ndash ; Barr virus ; ELISA ; enzyme-linked immunosorbent assay ; Fc¦ÃR ; Fc¦Ã receptor ; Fab-g ; IgG Fab fragment ; Fc-g ; IgG Fc fragment ; GAM ; goat anti mouse antibody ; HCMV ; human cytomegalovirus ; HSV
• 刊名：Journal of Immunological Methods
• 出版年：2013
• 出版时间：31 January, 2013
• 年：2013
• 卷：387
• 期：1-2
• 页码：21-35
• 全文大小：1282 K

文摘

IgG responses are crucial in antiviral defence and instrumental for the serodiagnosis of infections. Fc¦Ã receptors (Fc¦ÃRs), which recognize the Fc-part of IgG, differ regarding their IgG binding affinity, IgG subclass preference, cellular expression profile and pathogen elimination mechanisms elicited upon activation. Assessing their activation in vitro is of fundamental importance, but technically difficult. Therefore, a novel assay for measuring antiviral IgG antibodies triggering activation of individual host Fc¦Ã receptors was established. The assay comprises the co-cultivation of virus-infected target cells with immune IgG antibodies and mouse BW5147 hybridoma cells stably expressing chimeric Fc¦ÃR-CD3¦Æ chain molecules consisting of the extracellular domain of human Fc¦ÃRIIIA, Fc¦ÃRIIA or Fc¦ÃRI, respectively, fused to the transmembrane and intracellular domains of the mouse CD3¦Æ chain. Triggering of the chimeric Fc¦ÃR receptors by immune complexes formed on the surface of IgG-opsonized virus-infected target cells resulted in Fc¦ÃR activation leading to IL-2 secretion by BW5147 cells, which was quantified as a surrogate marker in an ELISA. Target cells infected with various human pathogenic viruses including herpes simplex virus type 1 (HSV-1), Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), measles virus (MV), and respiratory syncytial virus (RSV) or displaying human immunodeficiency virus-1 (HIV-1) gp120 evoke dose-dependent IgG responses demonstrating the universal applicability of the assay. Taken together, a new reliable and simple tool for measuring antibodies triggering activation of Fc¦Ã receptors was established. This assay will be instrumental for defining novel correlates of IgG immunity and the design of new therapeutic IgGs.