Characterization of in vitro antibody-dependent cell-mediated cytotoxicity activity of therapeutic antibodies — Impact of effector cells

Chung, Shan; Lin, Yuwen Linda; Reed, Chae; Ng, Carl; Cheng, Zhi-jie Jey; Malavasi, Fabio; Yang, Jihong; Quarmby, Valerie; Song, An

doi:10.1016/j.jim.2014.03.021

Cited by 49 publications

(38 citation statements)

References 38 publications

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“…TCZ showed no inhibitory effect on cytotoxicity. Chung et al 41 showed that NK cells are the dominant effector cells in PBMCs that mediate ADCC in their ADCC system where antibody-coated B lymphoblastoid cells was used as target cells, and only marginal ADCC activity was observed with monocytes. Major effector activity of NK cells in PBMC in ADCC was also reported by other study.…”

Section: Discussionmentioning

confidence: 99%

Tocilizumab (Anti-IL-6R) Suppressed TNFα Production by Human Monocytes in an In Vitro Model of Anti-HLA Antibody-Induced Antibody-Dependent Cellular Cytotoxicity

Shin

Mirocha

et al. 2017

Transplantation Direct

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BackgroundWe previously demonstrated that natural killer (NK) cells activated via FcγRIIIa (CD16) interactions with anti-HLA antibodies binding to peripheral blood mononuclear cells (PBMCs) in the in vitro antibody-dependent cellular cytotoxicity (ADCC) assay produced IFNγ. Here we investigate if other CD16 bearing cells are responsive to alloantigen via alloantibody in the in vitro ADCC and if the ADCC-induced cytokine reactions and cytotoxicity can be modified by the anti-interleukin 6 receptor (IL-6R) monoclonal antibody, Tocilizumab (TCZ).MethodsWhole blood from a normal individual was incubated overnight with irradiated allo-PBMCs pretreated with anti-HLA antibody positive (in vitro ADCC) or negative sera (mixed lymphocyte reaction [MLR]), with or without TCZ or control IgG. IFNγ+, TNFα+ or IL-6+ cell% in NK cells, monocytes and CD8+ T cells were enumerated by cytokine flow cytometry. ADCC using PBMCs (effector) and Farage B cells (FB, target) with anti-HLA antibody positive sera, with or without TCZ, was measured by flow cytometry.ResultsIFNγ+ and/or TNFα+ cell% in NK cells, monocytes and CD8+ T cells were elevated in the ADCC compared to the MLR condition. IL-6+ cells were significantly increased in ADCC versus MLR (10.2 ± 4.8% vs 2.7 ± 1.5%, P = 0.0003), but only in monocytes. TCZ treatment significantly reduced TNFα+ cell% in monocytes in ADCC, but had no effect on other cytokine+ cells. TCZ showed no effect on cytotoxicity in ADCC.ConclusionsIFNγ, TNFα, and IL-6 production induced by HLA antibody-mediated CD16 bearing cell activation in NK cells, monocytes, and CD8+ T cells suggests a potential role for ADCC and these inflammatory cytokines in mediation of antibody-mediated rejection. TCZ suppressed TNFα production in monocytes in the ADCC condition, suggesting a role of IL-6/IL-6R pathway in monocytes activation. Inhibition of this pathway could reduce the inflammatory cascade induced by alloantibody, although the inhibitory effect on cytotoxicity is minimal.

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Section: Discussionmentioning

confidence: 99%

Tocilizumab (Anti-IL-6R) Suppressed TNFα Production by Human Monocytes in an In Vitro Model of Anti-HLA Antibody-Induced Antibody-Dependent Cellular Cytotoxicity

Shin

Mirocha

et al. 2017

Transplantation Direct

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“…Thus, how Ag-bound Abs are presented for sensing by FcgRs underpins effector functions, such as Ab-dependent cellular cytotoxicity and Ab-dependent cellular phagocytosis, but is not simple to quantitate experimentally. Cell-based assays for such key cellular responses (40)(41)(42)(43)(44)(45) are difficult to standardize, so there is a need for an assay that simply evaluates the capacity of an immune complex (IC) to present Fcs for avid binding by FcgRs.…”

mentioning

confidence: 99%

Dimeric FcγR Ectodomains as Probes of the Fc Receptor Function of Anti-Influenza Virus IgG

Wines

Vanderven

Esparon

et al. 2016

The Journal of Immunology

100

130

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Ab-dependent cellular cytotoxicity, phagocytosis, and Ag presentation are key mechanisms of action of Abs arising in vaccine or naturally acquired immunity, as well of therapeutic mAbs. Cells expressing the low-affinity FcγRs (FcγRII or CD32 and FcγRIII or CD16) are activated for these functions when receptors are aggregated following the binding of IgG-opsonized targets. Despite the diversity of the Fc receptor proteins, IgG ligands, and potential responding cell types, the induction of all FcγR-mediated responses by opsonized targets requires the presentation of multiple Fc regions in close proximity to each other. We demonstrated that such “near-neighbor” Fc regions can be detected using defined recombinant soluble (rs) dimeric low-affinity ectodomains (rsFcγR) that have an absolute binding requirement for the simultaneous engagement of two IgG Fc regions. Like cell surface–expressed FcγRs, the binding of dimeric rsFcγR ectodomains to Ab immune complexes was affected by Ab subclass, presentation, opsonization density, Fc fucosylation, or mutation. The activation of an NK cell line and primary NK cells by human IgG-opsonized influenza A hemagglutinin correlated with dimeric rsFcγRIIIa binding activity but not with Ab titer. Furthermore, the dimeric rsFcγR binding assay sensitively detected greater Fc receptor activity to pandemic H1N1 hemagglutinin after the swine influenza pandemic of 2009 in pooled human polyclonal IgG. Thus these dimeric rsFcγR ectodomains are validated, defined probes that should prove valuable in measuring the immune-activating capacity of IgG Abs elicited by infection or vaccination or experimentally derived IgG and its variants.

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“…This discrepancy between in vitro and in vivo effects from the antiB16MOD immune sera was apparently due to the presence of different antibody isotypes, i.e., IgM, IgG1, and IgG2 (a + b), hampering for example the cytolytic activities of the first and the last ones. Another possibility is related to the IgFc receptors leading to the antibody-dependent cell cytotoxicity or ADCC [14,15]. Results obtained gave indications for a direct action relevant to different antibody isotype.…”

Section: Discussionmentioning

confidence: 92%

Characterization of effector components from the humoral and cellular immune response stimulated by melanoma cells exhibiting modified IGF-1 expression

Zhu

Trabado

Fan

et al. 2015

Biomedicine & Pharmacotherapy

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Characterization of in vitro antibody-dependent cell-mediated cytotoxicity activity of therapeutic antibodies — Impact of effector cells

Cited by 49 publications

References 38 publications

Tocilizumab (Anti-IL-6R) Suppressed TNFα Production by Human Monocytes in an In Vitro Model of Anti-HLA Antibody-Induced Antibody-Dependent Cellular Cytotoxicity

Tocilizumab (Anti-IL-6R) Suppressed TNFα Production by Human Monocytes in an In Vitro Model of Anti-HLA Antibody-Induced Antibody-Dependent Cellular Cytotoxicity

Dimeric FcγR Ectodomains as Probes of the Fc Receptor Function of Anti-Influenza Virus IgG

Characterization of effector components from the humoral and cellular immune response stimulated by melanoma cells exhibiting modified IGF-1 expression

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