γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay
γδ T cells are not MHC restricted, elicit cytotoxicity against various malignancies, are present in early post-transplant phases in novel stem cell transplantation (SCT) strategies and have been shown to mediate antibody-dependent cellular cytotoxicity (ADCC) with monoclonal antibodies (mAbs). These...
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doaj-c7f8799527b547a3905ad3778f11eded2020-11-24T22:52:51ZengFrontiers Media S.A.Frontiers in Immunology1664-32242014-12-01510.3389/fimmu.2014.00618118927γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assayUrsula Jördis Eva Seidel0Fabian eVogt1Fabian eVogt2Ludger eGrosse-Hovest3Ludger eGrosse-Hovest4Gundram eJung5Gundram eJung6Rupert eHandgretinger7Rupert eHandgretinger8Peter eLang9Peter eLang10University Children's Hospital TübingenUniversity of TübingenGerman Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ)University of TübingenSYNIMMUNE GmbHUniversity of TübingenGerman Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ)University Children's Hospital TübingenGerman Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ)University Children's Hospital TübingenGerman Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ)γδ T cells are not MHC restricted, elicit cytotoxicity against various malignancies, are present in early post-transplant phases in novel stem cell transplantation (SCT) strategies and have been shown to mediate antibody-dependent cellular cytotoxicity (ADCC) with monoclonal antibodies (mAbs). These features make γδ T cells promising effector cells for antibody-based immunotherapy in pediatric patients with B-lineage acute lymphoblastic leukemia (ALL). To evaluate combination of human γδ T cells with CD19 antibodies for immunotherapy of B-lineage ALL, γδ T cells were expanded after a GMP-compliant protocol and ADCC of both primary and expanded γδ T cells with an Fc optimized CD19 antibody (4G7SDIE) and a bispecific antibody with the specificities CD19 and CD16 (N19-C16) was evaluated in CD107a degranulation assays and intracellular cytokine staining (ICS). CD107a, TNF-α and IFN-γ expression of primary γδ T cells were significantly increased and correlated with CD16-expression of γδ T cells. γδ T cells highly expressed CD107a after expansion and no further increased expression by 4G7SDIE and N19-C16 was measured. Cytotoxicity of purified expanded γδ T cells targeting CD19-expressing cells was assessed in both europium-TDA release and in an impedance-based label-free method (using the xCELLigence system) measuring γδ T cell lysis in real-time. Albeit in the 2 h end-point europium-TDA release assay no increased lysis was observed, in real-time xCELLigence assays both significant antibody-independent cytotoxicity and ADCC of γδ T cells were observed. The xCELLigence system outperformed the end-point europium-TDA release assay in sensitivity and allows drawing of conclusions to lysis kinetics of γδ T cells over prolonged periods of time periods. Combination of CD19 antibodies with primary as well as expanded γδ T cells exhibits an promising approach, which may enhance clinical outcome of patients with pediatric B-lineage ALL and requires clinical evaluation.http://journal.frontiersin.org/Journal/10.3389/fimmu.2014.00618/fullADCCtherapeutic antibodiesTumor immunotherapyγδ T cell expansionxCELLigence system |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ursula Jördis Eva Seidel Fabian eVogt Fabian eVogt Ludger eGrosse-Hovest Ludger eGrosse-Hovest Gundram eJung Gundram eJung Rupert eHandgretinger Rupert eHandgretinger Peter eLang Peter eLang |
spellingShingle |
Ursula Jördis Eva Seidel Fabian eVogt Fabian eVogt Ludger eGrosse-Hovest Ludger eGrosse-Hovest Gundram eJung Gundram eJung Rupert eHandgretinger Rupert eHandgretinger Peter eLang Peter eLang γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay Frontiers in Immunology ADCC therapeutic antibodies Tumor immunotherapy γδ T cell expansion xCELLigence system |
author_facet |
Ursula Jördis Eva Seidel Fabian eVogt Fabian eVogt Ludger eGrosse-Hovest Ludger eGrosse-Hovest Gundram eJung Gundram eJung Rupert eHandgretinger Rupert eHandgretinger Peter eLang Peter eLang |
author_sort |
Ursula Jördis Eva Seidel |
title |
γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay |
title_short |
γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay |
title_full |
γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay |
title_fullStr |
γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay |
title_full_unstemmed |
γδ T cell-mediated antibody-dependent cellular cytotoxicity with CD19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay |
title_sort |
γδ t cell-mediated antibody-dependent cellular cytotoxicity with cd19 antibodies assessed by an impedance-based label-free real-time cytotoxicity assay |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2014-12-01 |
description |
γδ T cells are not MHC restricted, elicit cytotoxicity against various malignancies, are present in early post-transplant phases in novel stem cell transplantation (SCT) strategies and have been shown to mediate antibody-dependent cellular cytotoxicity (ADCC) with monoclonal antibodies (mAbs). These features make γδ T cells promising effector cells for antibody-based immunotherapy in pediatric patients with B-lineage acute lymphoblastic leukemia (ALL). To evaluate combination of human γδ T cells with CD19 antibodies for immunotherapy of B-lineage ALL, γδ T cells were expanded after a GMP-compliant protocol and ADCC of both primary and expanded γδ T cells with an Fc optimized CD19 antibody (4G7SDIE) and a bispecific antibody with the specificities CD19 and CD16 (N19-C16) was evaluated in CD107a degranulation assays and intracellular cytokine staining (ICS). CD107a, TNF-α and IFN-γ expression of primary γδ T cells were significantly increased and correlated with CD16-expression of γδ T cells. γδ T cells highly expressed CD107a after expansion and no further increased expression by 4G7SDIE and N19-C16 was measured. Cytotoxicity of purified expanded γδ T cells targeting CD19-expressing cells was assessed in both europium-TDA release and in an impedance-based label-free method (using the xCELLigence system) measuring γδ T cell lysis in real-time. Albeit in the 2 h end-point europium-TDA release assay no increased lysis was observed, in real-time xCELLigence assays both significant antibody-independent cytotoxicity and ADCC of γδ T cells were observed. The xCELLigence system outperformed the end-point europium-TDA release assay in sensitivity and allows drawing of conclusions to lysis kinetics of γδ T cells over prolonged periods of time periods. Combination of CD19 antibodies with primary as well as expanded γδ T cells exhibits an promising approach, which may enhance clinical outcome of patients with pediatric B-lineage ALL and requires clinical evaluation. |
topic |
ADCC therapeutic antibodies Tumor immunotherapy γδ T cell expansion xCELLigence system |
url |
http://journal.frontiersin.org/Journal/10.3389/fimmu.2014.00618/full |
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