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酵素抗体法を用いた菌体内コレラ菌毒素の電顕的観察


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Title: 酵素抗体法を用いた菌体内コレラ菌毒素の電顕的観察
Other Titles: Observation of Cholera Toxin in Vibrio cholerae by Using Enzyme Labeled Antibody
Authors: 岩永, 正明 / 江原, 雅彦 / 内藤, 達郎
Authors (alternative): Iwanaga, Masaaki / Ehara, Masahiko / Naito, Tatsuro
Issue Date: 30-Mar-1981
Publisher: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
Citation: 熱帯医学 Tropical medicine 23(1). p13-21, 1981
Abstract: 毒素産生と非産生の条件下で培養したコレラ菌569B株および毒素非産生株K25の電顕形態学的比較に引き続き,酵素標識抗毒素抗体を用いて菌体内における毒素の局在を観察しようとした.使用菌株および2種の培養条件は既報と同様である.choletoxで免疫して得た家兎血清(PHA価10,240倍)よりIgGを硫安塩析とDEAE-sephadex A-50ゲル濾過で精製,濃縮(同5,120倍),これにAvrameas and Ternynck (1971)の方法でhorseradish peroxidase (Sigma RZ: 3.1)を結合させた(同1,024倍),洗浄菌体を0.5%グルタールアルデヒドで固定,洗浄後HRP-IgGを,再洗浄後0.2% diaminobenzidine,ついで過酸化水素を作用させた.充分洗浄後1%オスミウム酸で固定,常法で脱水,エポン包埋,超薄切し,一部では酢酸ウラニルとクエン酸鉛染色を行って,鏡検した.毒素産生条件下の569Bでは,全般的に大円型細胞よりも桿状細胞に毒素が証明され,後者ではperiplasmic space,細胞膜,メソゾーム様構造の部分,前者では空胞様部分の内側表層に認められた.毒素の存在様式は,1)大部分細胞内で少量が表層,2)内部にもあるが大量が細胞表面,3)periplasmic spaceになく細胞表面,の3種にまとめられた.前報で毒素産生条件にのみ多くみられた,細胞壁の突出部とこれが遊離した粒子,また対照(非結合HRPの使用,抗毒素IgG前処置,K25株)ではすべて毒素染出はみられなかった. / Pleomorphic changes were previously observed in Vibrio cholerae during toxin production in the syncase medium. These changes are mainly characterized by large round cells, and the release of membrane-enveloped particles following budding and pinching off of the cell walls. To understand the relationship between toxin production and these morphological changes, localization of the toxin in the organism was examined using horseradish peroxidase labeled antibody. Toxin was detected mainly in the periplasmic space of rod shape organism, and occasionally in large round cells, but not in the membraneenveloped particles. Toxin was also found in the mesosomal structures and outer surface materials in some of these cells. Besides, a densely stained, branching network was observed first time in V. cholerae without the fixation with glutaraldehyde.
URI: http://hdl.handle.net/10069/4294
ISSN: 03855643
Type: Departmental Bulletin Paper
Appears in Collections:Volume 23, No. 1

Citable URI : http://hdl.handle.net/10069/4294

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