백일해균의 세포벽과 원형질의 항원성에 관한 연구

Abstract

Many investigators have reported that B. pertussis cells possess protective antigen, histamine sensitizing factor, heat stable and labile toxin, hemagglutinin, agglutinogen and the others. However, the location in the cell of the active material is not known with certainly. The present experiments were made to determine the antigenic structure of B. pertussis. In this study, the method of preparation of cell wall, protoplasm and whole cell of B. pertussis was described and the protective antigenicity studied by phagocytic rate of peritoneal macrophage obtained from immune or control mice, histamine sensitizing activity and toxicity of these preparations of B. pertussis were presented. Material and Method: 1) Strains of B. pertussis: Laboratory stock strains, routinely used for vaecine preparation in N.1. H. Korea, were employed 2) Media and Culture: Culture on Bordet Gengou Agar containing 20% defibrinated sheep blood was incubated at 370 c for 2 days. The growth from plate was suspended in cold physical saline. 3) The method of obtaining of cell wall and protoplasm: The preparations of cell wall and protoplasm from these cells were followed, with mino modification, the method previously described by Munoz (1959) an d Ribi (1959) 4) Protective antigenicity by phagocytic rate of immune or control macrophage: Following immunizing injections of whole cell, cell wall and protoplasm of B. pertusis, peritoneal exudates were induced in the immunized and control mice by intra peritoneal injection of sterile glycogen. Collected macrophages were infected by cells of B. pertussis and then the mixtures were incubated to allow phagocytosis. Phagocytic rate was based on the number of intra cellular bacterial units per 100 macrophages counted in random field. 5) Histamine sensitizing test: Suspensions of whole cell, cell wall and solutions of protoplasm were made in physiological saline, and then 5 fold dilutions were made in physiological saline. Groups of 10 mice were challenged (I. P.) with 0.5mg of histamine base 4 days after the sensitizing injection. 6) Toxicity test: a) Each preparation was weighed and resuspended in lOml of physiological saline, 5 fold dilutions were then made. Groups of 10 mice each were injected intraperitoneally with Iml of the various concentrations of the three different preparations. LD 50 values of these fractions were determined in mice (12±lgm). The results were calculated according to the method of Reed Muench after 3 days observation. b) Suspensions of these fractions were adjusted by Klett summerson photoelectriccolorimeter and then 0.2 ml of these suspensions treated at various temperature was injected intra dermally into lateral dorsal surface of clipped normal rabbit. The results were summerized as follows: 1) Protective antigen of B. pertussis investigated by immune or control macrophage in the presence of immune or control serum was mainly located in cell wall preparation. 2) Histamine sensitizing factor of B. pertussis was mainly located in cell wall preparation. 3) Heat labile toxin of B. pertussis was mainly present in the protoplasm preparation.