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Tove Christensen

Flow cytometric assay detecting cytotoxicity against human endogenous retrovirus antigens expressed on cultured multiple sclerosis cells

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Flow cytometric assay detecting cytotoxicity against human endogenous retrovirus antigens expressed on cultured multiple sclerosis cells. / Møller-Larsen, A; Brudek, T; Petersen, T; Petersen, E L; Aagaard, M; Hansen, Dorte; Christensen, T.

I: Clinical and Experimental Immunology, Bind 173, Nr. 3, 09.2013, s. 398-410.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Møller-Larsen, A ; Brudek, T ; Petersen, T ; Petersen, E L ; Aagaard, M ; Hansen, Dorte ; Christensen, T. / Flow cytometric assay detecting cytotoxicity against human endogenous retrovirus antigens expressed on cultured multiple sclerosis cells. I: Clinical and Experimental Immunology. 2013 ; Bind 173, Nr. 3. s. 398-410.

Bibtex

@article{5219829364a34b4891960fe6d28245e5,
title = "Flow cytometric assay detecting cytotoxicity against human endogenous retrovirus antigens expressed on cultured multiple sclerosis cells",
abstract = "Damage of target cells by cytotoxicity, either mediated by specific lymphocytes or via antibody-dependent reactions, may play a decisive role in causing the central nervous system (CNS) lesions seen in multiple sclerosis (MS). Relevant epitopes, antibodies towards these epitopes and a reliable assay are all mandatory parts in detection and evaluation of the pertinence of such cytotoxicity reactions. We have adapted a flow cytometry assay detecting CD107a expression on the surface of cytotoxic effector cells to be applicable for analyses of the effect on target cells from MS patients expressing increased amounts of human endogenous retrovirus antigens. MS patients also have increased antibody levels to these antigens. The target cells are spontaneously growing peripheral blood mononuclear cells (PBMCs) of B cell lineage, expressing human endogenous retrovirus HERV epitopes on their surface. Polyclonal antibodies against defined peptides in the Env- and Gag-regions of the HERVs were raised in rabbits and used in antibody-dependent cell-mediated cytotoxicity (ADCC) -assays. Rituximab{\textregistered} (Roche), a chimeric monoclonal antibody against CD20 expressed primarily on B cells, was used as control antibody. Without antibodies this system is suitable for analyses of natural killer cell activity. In optimization of the assay we have used effector lymphocytes from healthy donors. The most effective effector cells are CD56(+) cells. CD8(+) T cells also express CD107a in ADCC. Using the adapted assay, we demonstrate significant ADCC activity to target cells expressing HERV epitopes, and additionally a low level of NK activity.",
keywords = "Adult, Antibodies, Antibodies, Monoclonal, Antibodies, Monoclonal, Murine-Derived, Antibody-Dependent Cell Cytotoxicity, Antigens, Cells, Cultured, Cytotoxicity, Immunologic, Endogenous Retroviruses, Female, Flow Cytometry, Humans, Killer Cells, Natural, Male, Middle Aged, Multiple Sclerosis, T-Lymphocytes, Cytotoxic, Young Adult",
author = "A M{\o}ller-Larsen and T Brudek and T Petersen and Petersen, {E L} and M Aagaard and Dorte Hansen and T Christensen",
note = "{\textcopyright} 2013 British Society for Immunology.",
year = "2013",
month = sep,
doi = "10.1111/cei.12133",
language = "English",
volume = "173",
pages = "398--410",
journal = "Clinical and Experimental Immunology",
issn = "0009-9104",
publisher = "Wiley-Blackwell Publishing Ltd.",
number = "3",

}

RIS

TY - JOUR

T1 - Flow cytometric assay detecting cytotoxicity against human endogenous retrovirus antigens expressed on cultured multiple sclerosis cells

AU - Møller-Larsen, A

AU - Brudek, T

AU - Petersen, T

AU - Petersen, E L

AU - Aagaard, M

AU - Hansen, Dorte

AU - Christensen, T

N1 - © 2013 British Society for Immunology.

PY - 2013/9

Y1 - 2013/9

N2 - Damage of target cells by cytotoxicity, either mediated by specific lymphocytes or via antibody-dependent reactions, may play a decisive role in causing the central nervous system (CNS) lesions seen in multiple sclerosis (MS). Relevant epitopes, antibodies towards these epitopes and a reliable assay are all mandatory parts in detection and evaluation of the pertinence of such cytotoxicity reactions. We have adapted a flow cytometry assay detecting CD107a expression on the surface of cytotoxic effector cells to be applicable for analyses of the effect on target cells from MS patients expressing increased amounts of human endogenous retrovirus antigens. MS patients also have increased antibody levels to these antigens. The target cells are spontaneously growing peripheral blood mononuclear cells (PBMCs) of B cell lineage, expressing human endogenous retrovirus HERV epitopes on their surface. Polyclonal antibodies against defined peptides in the Env- and Gag-regions of the HERVs were raised in rabbits and used in antibody-dependent cell-mediated cytotoxicity (ADCC) -assays. Rituximab® (Roche), a chimeric monoclonal antibody against CD20 expressed primarily on B cells, was used as control antibody. Without antibodies this system is suitable for analyses of natural killer cell activity. In optimization of the assay we have used effector lymphocytes from healthy donors. The most effective effector cells are CD56(+) cells. CD8(+) T cells also express CD107a in ADCC. Using the adapted assay, we demonstrate significant ADCC activity to target cells expressing HERV epitopes, and additionally a low level of NK activity.

AB - Damage of target cells by cytotoxicity, either mediated by specific lymphocytes or via antibody-dependent reactions, may play a decisive role in causing the central nervous system (CNS) lesions seen in multiple sclerosis (MS). Relevant epitopes, antibodies towards these epitopes and a reliable assay are all mandatory parts in detection and evaluation of the pertinence of such cytotoxicity reactions. We have adapted a flow cytometry assay detecting CD107a expression on the surface of cytotoxic effector cells to be applicable for analyses of the effect on target cells from MS patients expressing increased amounts of human endogenous retrovirus antigens. MS patients also have increased antibody levels to these antigens. The target cells are spontaneously growing peripheral blood mononuclear cells (PBMCs) of B cell lineage, expressing human endogenous retrovirus HERV epitopes on their surface. Polyclonal antibodies against defined peptides in the Env- and Gag-regions of the HERVs were raised in rabbits and used in antibody-dependent cell-mediated cytotoxicity (ADCC) -assays. Rituximab® (Roche), a chimeric monoclonal antibody against CD20 expressed primarily on B cells, was used as control antibody. Without antibodies this system is suitable for analyses of natural killer cell activity. In optimization of the assay we have used effector lymphocytes from healthy donors. The most effective effector cells are CD56(+) cells. CD8(+) T cells also express CD107a in ADCC. Using the adapted assay, we demonstrate significant ADCC activity to target cells expressing HERV epitopes, and additionally a low level of NK activity.

KW - Adult

KW - Antibodies

KW - Antibodies, Monoclonal

KW - Antibodies, Monoclonal, Murine-Derived

KW - Antibody-Dependent Cell Cytotoxicity

KW - Antigens

KW - Cells, Cultured

KW - Cytotoxicity, Immunologic

KW - Endogenous Retroviruses

KW - Female

KW - Flow Cytometry

KW - Humans

KW - Killer Cells, Natural

KW - Male

KW - Middle Aged

KW - Multiple Sclerosis

KW - T-Lymphocytes, Cytotoxic

KW - Young Adult

U2 - 10.1111/cei.12133

DO - 10.1111/cei.12133

M3 - Journal article

C2 - 23656307

VL - 173

SP - 398

EP - 410

JO - Clinical and Experimental Immunology

JF - Clinical and Experimental Immunology

SN - 0009-9104

IS - 3

ER -