RRC ID 56580
Author Hayashi Y, Okutani M, Ogawa S, Tsukahara T, Inoue R.
Title Generation of anti-porcine CD69 monoclonal antibodies and their usefulness to evaluate early activation of cellular immunity by flow cytometric analysis.
Journal Anim Sci J
Abstract T cell-mediated cellular immunity and humoral immunity are equally important for the prevention of diseases. To assess activation of human and mouse cellular immunity, early activation markers of lymphocytes are often used in flow cytometry targeting expression of CD69 molecules. Response of humoral immunity against infection or vaccination has been well investigated in pigs, but that of cellular immunity has been largely neglected due to lack of direct evaluation tools. Thus, in pig research a proper assay of antibody reacted with porcine CD69 is still unavailable. In the present study, two anti-porcine CD69 mAb-producing mouse hybridomas, 01-14-22-51 (IgG2b-κ) and 01-22-44-102 (IgG2a-κ), both showing fine reactivity with phorbol 12-myristate 13-acetate (PMA) and ionomycin-stimulated porcine peripheral blood lymphocytes in flow cytometry, were established. When porcine peripheral blood lymphocytes were activated with PMA and ionomycin and analyzed by flow cytometry, it was found that both mAbs generated in this study stained about 70% of lymphocytes. In contrast, after an identical procedure, only 5% and 13.5% of lymphocytes were stained with anti-interferon-γ mAb and anti-tumor necrosis factor-α mAb, respectively. These results indicate that evaluation of cellular immunity activation turns more sensitive after using our newly generated mAbs.
Volume 89(5)
Pages 825-832
Published 2018-5-1
DOI 10.1111/asj.12989
PMID 29460471
MeSH Animals Antibodies, Monoclonal* Antigens, CD / immunology* Antigens, Differentiation, T-Lymphocyte / immunology* Flow Cytometry / methods* Humans Immunity, Cellular / immunology* Ionomycin / immunology Lectins, C-Type / immunology* Mice Swine T-Lymphocytes / immunology* Tetradecanoylphorbol Acetate / analogs & derivatives Tetradecanoylphorbol Acetate / immunology
IF 1.301
Times Cited 1
Human and Animal Cells SP2/0-Ag14(RCB0209)