Bacterial growth and virulence factors production by different Bordetella pertussis strains.

The aim of this study was to compare bacteria growth and pertussis antigens (pertussis toxin-PT, filamentous haemagglutinin-FHA and endotoxin-LPS) production by 11 Bordetella pertussis strains. A synthetic Stainer-Scholte culture medium supplemented with (2,6-0-dimethyl) beta-cyclodextrin (heptakis) or methylcellulose (for greater PT and FHA production) and solid modified Cohen-Wheeler medium (for LPS isolation) were used. Our results demonstrated that heptakis and methylcellulose were more effective for antigens production than for bacterial growth. It was interesting that these stimulated substances supported the bacterial growth from small inocula. The investigated strains differed in PT, FHA and LPS production. The best PT producer was the B. pertussis 134 strain, the worst B. pertussis 2897. The differences in FHA production are not as big as in PT production, but the B. pertussis 2897 was also the worst FHA producer. Isolated LPS consists of dry bacteria pellet ranging from 1,9% (B.p. Tohama) to 5,6% (B.p. 3803 strain). No great differences in serological activity of LPS isolated from different strains were observed. In the haemagglutination inhibition test the endotoxin isolated from B.p. 509 and B.p. Tohama strains showed the highest activity.

[Properties of an acellular preparation of Bordetella pertussis and its analysis after centrifugation in saccharose gradient]. / Wlasciwosci bezkomórkowego preparatu Bordetella pertussis i jego analiza po wirowaniu w gradiencie sacharozy.

Acellular preparations were studied which were obtained by a method of acidic extraction from culture of strain Tohama of Bordetella pertussis. Obtained preparations EM I and EM II were exhibiting property of leukocytosis stimulation and were respectively of following values: 43.5 and 19 u LPF/mg of protein. They also caused hemagglutination (150 and 19 u HA/mg of protein). Both preparations were exhibiting protective effect in intracerebral test performed on mice. After differentiation of EM I preparation by preparative centrifugation in saccharose gradient two fractions were obtained, of which lighter was exhibiting high hemagglutinating activity and leukocytosis stimulation (198 u LPF/mg of protein). Heaver fraction contained mainly endotoxin. Applied method permitted for partial purification of acellular preparation of B. pertussis; mainly decrease in its content of endotoxin responsible for high reactogenicity of pertussis vaccines.

A versatile method for purification of diphtheria toxoid for immunization purposes.

Diphtheriae growth supernatant. The toxoid was quantitatively recovered from the immunosorbent in two fractions: the first in 0.005 M hydochloric acid and the second in 6' M urea containing 0.1 M HCL. Both fractions were soluble in aqueous solvents, and both had a similar specific activity: between 200 and 250 Lf/mg protein, and about 1.5 protective unit (ED50) per l Lf unit, as assayed in guinea pigs. Using the same procedure, the specific activity of a commerical toxoid preparation, purified and concentrated, was increrased two fold, from 130 Lf/mg to 260 Lf/mg protein.