31 research outputs found
A Study on the Antigenic Activities of Cell Wall and Protoplasm of B. pertussis.
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
A Study on the soluble antigen of Bordetella pertussis
It was well established that the inoculation of pertussis vaccine was capable of reducing the incidence
of whooping cough.
However, complete elimination of toxicity of pertussis
vaccine was not succeeded yet.
Thus this experiment was carried out to reduce the
toxicity of pertussis vaccine.
This paper described the conditions under which
sodium desoxycholate may be used to liberate the the
soluble protective antigen in nontoxic form and the
potency of protective antigen liberated from both 48
hrs. and 72 hrs. pertussis cultures.
Results were as follows:
1. The liberation of soluble protective antigen from
pertussis cells in non-toxic form was possible using
both optimal concentration (0.5%) and pH(8.5)
of sodium desoxycholate solution.
2. The protective antigen liberated from 48hrs. pertussis
culture at 0.5% sodium desoxycholate solution
exhibited a greater degree of potency than that
liberated from 72hrs. cultures
