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- W2078485984 abstract "To~i,con, Supp1.3, pp.203-206, 1983. Pergamon Press Ltd.Printed in Great Britain.CHEMILUMINESCENCE RESPONSE OF PHAGOCYTES TO BACTERIAL TOXINSS. Kanegasaki, T. Tamita, K. Momoi, M. Noda and I . KatoInst. of Med. Sci., Univ. of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, JapanINTRODUCTIONBinding of particulates to the membrane of phagocytes such as polymorphonuclearleukocytes (PMNs) and mononuclear phagocytes is known to burst the oxidative metabo-lism of these cells. In consequence, the cells produce substantial quantities of activeoxygens such as superoxide and hydrogen peroxide, and possibly singlet oxygen and hydroxyradicals. These active oxyges are believed to play an essential role in killing in-fectious agents as well as neoplastic cells. Production of oxidative radicals can beassayed by chemiluminescence generation.Certain soluble agents such as digitonin, fatty acids, fluoride and phorbolrt~yristateacetate, which affect m~nbrane of phagocytes, are known to induce respiratory burst. Wereported previously that bacterial exotoxins including staphylococcal alpha toxin anddiphtheria toxin stimulated human and marine phagocytes and induced chemiluminescence inthe presence of luminol (KANEGASAKI et ae, 1980, MOOKERJEE et a.C., 1982). The resultindicates that these toxins, though different in their specific action mechanism, prima-rily affect the membrane of phagocytes and induce active oxygen production. Since cyto-plasmic membrane is a primary target for cytotoxins, other bacterial toxins are alsoexpected to stimulate phagocytes. If this is so, chemiluminescence assay can generallybe used for qualitative and quantitative tests for the toxins. For this purpose, wefurther investigated the kinetics of light emission from human, rabbit and marine PMNsupon exposure to various toxins.MATERIALS AND METHODSRESULTSComparri.eon ob f~inetica o~ che~ni,fuMinedcence .induced 6y vaninue .towind.203Towi,naHighly purified diphtheria toxin (KATO et af., 1960), alpha toxin (WATANABE b KATO,1974), and leukocidin (NODA ~t u.L., 1980x) were used. Delta toxin was purified twoways according to the methods described by Neatly et of., 1980 and Smith 8 Shaw, 1981.Cholera toxin was purchased from Kaketsuken, Kumamoto.PhagocytesHuman and rabbit PMNs were obtained from peripheral blood and isolated using lympho-prep (Nyegaad, Oslo) and dextran (B~YUM, 1968). Mmonium chloride solution was used forerythrocyte lysis. The final suspension was more than 95~ pure for granulocytes.Marine exudate PMNs were obtained from C3H mouse peritoneal cavity 12 hours afterinjection of casein.Chen~i,tun~ineacence mectbWrernentCh~niluminescence was measured at 37°C using chemiluminescent analyser, Biolu~atLB 9500 (Lab. Prof. Dr. Berthold, Widbad). To plastic vials containing 1 .6 x 10 cellsin 1 ml, 40 u1 of luminol solution (40 ug/ml) was added and background counts wererecorded. The experiments were started by the addition of toxins and the time courseof photon count rate was traced.Effects of diphtheria toxin, staphylococcal alpha and delta toxins, staphylococcalleukocldin and cholera toxin on chemiluminescence emission from human PMNs were compared.As shown in Fig. 1, these toxins all enhanced chemiluminescence generation, although theinduction by cholera toxin was less prominent as compared to other toxins tested.Immediate light emission was observed upon exposure to delta toxin with the maximalresponse approximately 1 min later, followed by rapid subsidence. Chemiluminescence" @default.
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- W2078485984 date "1983-01-01" @default.
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- W2078485984 title "Chemiluminescence response of phagocytes to bacterial toxins" @default.
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- W2078485984 doi "https://doi.org/10.1016/0041-0101(83)90191-5" @default.
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