中毒休克综合征毒素-1在小鼠金黄色葡萄球菌乳腺炎中的作用
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摘要
为探究金黄色葡萄球菌(S.aureus)超抗原毒素在其产生菌引起的乳腺炎中的作用,本研究纯化制备了重组中毒休克综合征毒素1(rTSST-1),并以5μg、15μg和30μg 3个剂量接种小鼠乳腺,结果显示3个剂量均引起了小鼠乳腺眼观炎症变化,并随着感染剂量加大病变呈现加重趋势。以1×10~2cfu和1×10~3cfu两个剂量的S.aureus R8菌株感染乳腺后24h眼观均出现了红肿变化,48h、72h时出现了化脓、干酪样坏死,且高剂量组病变重于低剂量组。进一步比较研究了以rTSST-1(15μg)、S.aureus R8(1×102cfu)和rTSST-1(15μg)+S.aureus R8(1×102cfu)等接种小鼠乳腺后对乳腺的影响,结果显示rTSST-1组小鼠乳腺眼观病变最轻,S.aureus R8组最重;接种24h后观察组织病变,结果显示rTSST-1组与rTSST-1+S.aureus R8组病变相近,小鼠乳腺腺泡形态边缘不完整、腺泡内有大量嗜中性粒细胞浸润。然而S.aureus R8组小鼠乳泡形态边缘破坏严重但嗜中性粒细胞等免疫细胞浸润数量明显少于前两组。乳腺活菌计数结果显示,S.aureus R8组小鼠的活菌数显著高于rTSST-1+S.aureus R8组。本研究结果显示TSST-1可单独引起小鼠乳腺炎症,而在TSST-1与S.aureus同时感染乳腺时,TSST-1减轻了乳腺组织的炎症程度。这一结果显示TSST-1引起的小鼠腺泡中嗜中性粒细胞的大量浸润可能对促进乳腺中S.aureus的有效清除具有重要作用。本研究将为有效防控S.aureus引起的奶牛乳腺炎提供新的参考依据。
To investigate the pathogenesis of staphylococcal superantigenic toxins in mastitis induced by Staphylococcus aureus(S.aureus), recombinant superantigenic toxin, toxic shock syndrome toxin-1(rTSST-1), was prepared and injected into mice mammaries(5, 15 and 30μg per mammary). The results showed that r TSST-1 induced inflammatory responses in mammaries as a dose dependent manner. The mammaries infected with S.aureus R8 at the doses of 1 ×10~2 and 1 ×10~3cfu were significantly inflamed at 24h, and then showed festering and caseous necrosis. The mammaries infected with S.aureus R8 plus rTSST-1 together showed the slighter inflammation than that of the mammaries infected with S.aureus R8 alone. Bacteria counts in the mammaries infected with S.aureus R8 plus rTSST-1 were significantly lower than that with S.aureus R8 only. In addition,neutrophil infiltrated numbers in gland bubbles of the mammaries infected with S.aureus R8 plus rTSST-1 were significantly higher than that of infected with S.aureus R8 alone. These results indicated that TSST-1 itself could induce mastitis slightly, and also could alleviate S.aureus-induced inflammation response in mammaies when TSST-1 was injected with S.aureus together,suggesting that TSST-1-increased neutrophil infiltration could be important for clearances of bacteria in mammary. Our research could provide new reference basis for developing efficient control methods against cow mastitis caused by S.aureus.
To investigate the pathogenesis of staphylococcal superantigenic toxins in mastitis induced by Staphylococcus aureus(S.aureus), recombinant superantigenic toxin, toxic shock syndrome toxin-1(rTSST-1), was prepared and injected into mice mammaries(5, 15 and 30μg per mammary). The results showed that r TSST-1 induced inflammatory responses in mammaries as a dose dependent manner. The mammaries infected with S.aureus R8 at the doses of 1 ×10~2 and 1 ×10~3cfu were significantly inflamed at 24h, and then showed festering and caseous necrosis. The mammaries infected with S.aureus R8 plus rTSST-1 together showed the slighter inflammation than that of the mammaries infected with S.aureus R8 alone. Bacteria counts in the mammaries infected with S.aureus R8 plus rTSST-1 were significantly lower than that with S.aureus R8 only. In addition,neutrophil infiltrated numbers in gland bubbles of the mammaries infected with S.aureus R8 plus rTSST-1 were significantly higher than that of infected with S.aureus R8 alone. These results indicated that TSST-1 itself could induce mastitis slightly, and also could alleviate S.aureus-induced inflammation response in mammaies when TSST-1 was injected with S.aureus together,suggesting that TSST-1-increased neutrophil infiltration could be important for clearances of bacteria in mammary. Our research could provide new reference basis for developing efficient control methods against cow mastitis caused by S.aureus.
引文
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[10]马保臣,柴同杰,秦卓明,等.用PCR与ELISA、RPLA对照检测乳腺炎金黄色葡萄球菌肠毒素和毒素休克综合征毒素-1[J].中国兽医学报,2008,28(08):901-905.
[11]王新,韦艺媛,张静,等.乳房炎奶牛金黄色葡萄球菌毒素基因的检测及PFGE分型研究[J].畜牧兽医学报,2011,42(07):974-980.
[12]Cui Jing-chun,Hu Dong-liang,Lin Yan-chun,et al.Immunization with glutathione S-transferase and mutant toxic shock syndrome toxin 1 fusion protein protects against Staphylococcus aureus infection[J].FEMS Immunol Med Micorbiol,2005,45:45-51.
[13]张宝君,崔京春,张献,等.金葡菌乳腺感染模型的建立与评价[J].中国预防兽医学报,2009,31(05):365-369.
[14]Holtfreter S,Broker B M.Staphylococcal superantigens:do they play a role in sepsis?[J].Arch Immunol Ther Exp,2005,53(1):13-27.
[15]冯乐平,陈军,王海.超抗原SEB对淋巴细胞抗肿瘤免疫效果的影响[J].实用肿瘤学杂志,2001,15(1):1-2.
[16]李明,邢飞跃,李岩.超抗原SEB或SEC对T细胞增殖和活化的影响[J].实用医学杂志,2005,21(15):1612-1614.
[2]尹荣兰,李琳,李昱洁,等.奶牛乳腺炎金黄色葡萄球菌新型疫苗研究进展[J].吉林畜牧兽医,2011,(12):5-8.
[3]Katsuda K,Hata E,Kobayashi H,et al.Molecular typing of Staphylococcus aureus isolated from bovine mastitic milk on the basis of toxin genes and coagulase gene polymorphisms[J].Vet Microbiol,2005,105(03-04):301-305.
[4]孙颖,汤婷婷,鲍庆晗,等.牛乳金黄色葡萄球菌分离株超抗原肠毒素基因分型研究[J].动物医学进展,2014,(12):14-18.
[5]Hu Dong-liang,Cui Jing-chun,Omoe K,et al.A mutant of staphylococcal enterotoxin C devoid of bacterial superantigenic activity elicits a Th2 immune response for protection against Staphylococcus aureus infection[J].Infect Immun,2005,73(01):174-180.
[6]Hu Dong-liang,Omoe K,ShimodaY.et al.Induction of emetic response to staphylococcal enterotoxins in the house musk shrew(Suncus murinus)[J].Infect Immun,2003,71:567-570.
[7]Omoe K,Hu Dong-liang,Takahashi O H,et al.Identification and characterization of a new staphylococcal enterotoxin-related putative toxin encoded by two kinds of plasmids[J].Infect Immun,2003,71(10):6088-6094.
[8]胡东良,品川邦汎,刘佩红,等.动物性食品中金黄色葡萄球菌产肠毒素的研究[J].中国食品卫生学杂志,1996,8(03):9-15.
[9]蒋长苗,刘佩紅,丁建平,等.中国の动物性食品における黄色ブドウ球菌汚染と分离菌株のエンテロトキシン产生性[J].日本食品微生物学杂志,2001,18:43-47.
[10]马保臣,柴同杰,秦卓明,等.用PCR与ELISA、RPLA对照检测乳腺炎金黄色葡萄球菌肠毒素和毒素休克综合征毒素-1[J].中国兽医学报,2008,28(08):901-905.
[11]王新,韦艺媛,张静,等.乳房炎奶牛金黄色葡萄球菌毒素基因的检测及PFGE分型研究[J].畜牧兽医学报,2011,42(07):974-980.
[12]Cui Jing-chun,Hu Dong-liang,Lin Yan-chun,et al.Immunization with glutathione S-transferase and mutant toxic shock syndrome toxin 1 fusion protein protects against Staphylococcus aureus infection[J].FEMS Immunol Med Micorbiol,2005,45:45-51.
[13]张宝君,崔京春,张献,等.金葡菌乳腺感染模型的建立与评价[J].中国预防兽医学报,2009,31(05):365-369.
[14]Holtfreter S,Broker B M.Staphylococcal superantigens:do they play a role in sepsis?[J].Arch Immunol Ther Exp,2005,53(1):13-27.
[15]冯乐平,陈军,王海.超抗原SEB对淋巴细胞抗肿瘤免疫效果的影响[J].实用肿瘤学杂志,2001,15(1):1-2.
[16]李明,邢飞跃,李岩.超抗原SEB或SEC对T细胞增殖和活化的影响[J].实用医学杂志,2005,21(15):1612-1614.