Pertussis toxin glutaraldehyde

The effects of different inactivating agents on the biological activity of the histamine sensitization factor of Bordetella pertussis toxin were examined. The agents were used for inactivation in the preparation of whole cell pertussis suspension. The histamine sensitizing activity was reduced to 36.9-13.3% by treatment with glutaraldehyde, to about 50% by treatment with formaldehyde and by the acetone-II treatment, relative to the reduction by heat treatment. Treatment with thimerosal and the acetone-I treatment did not reduce the histamine sensitizing activity as the 50% histamine sensitizing doses of the heat inactivated pertussis preparation, the thimerosal inactivated pertussis preparation and the acetone-I treated pertussis preparation were very similar...

Endotoxin protein represents a group of immunobiologically active p polypeptides which are associated with the lipopolysaccharide endotoxin in the outer membrane of Gram-negative bacteria. To study the adjuvant effect of endotoxin protein, CF-1 mice were immunized intraperitoneally with graded doses of glutaraldehyde-inactivated cholera toxoid with and without endotoxin protein prepared from Bordetella pertussis, Salmonella typhi or Vibrio cholerae. Immune responsiveness was assessed by measuring resistance to intravenous challenge with cholera enterotoxin and by serum antitoxin responses...

Pertussigen [pertussis toxin (Ptx)] from Bordetella pertussis, when detoxified, induces protection in mice to intracerebral challenge (ic) with virulent B. pertussis. In its native form, minute nonprotective doses promote the development of immunity induced by other antigens of B. pertussis. As little as 4 ng of Ptx, given with a nonprotective dose of 8 X 10(7) killed cells of the phase III Sakairi strain, promoted detectable protection to ic challenge. Native Ptx in doses of 0.4 to 400 ng did not protect mice, and vaccines made from strains not producing Ptx induced only weak protection...

Treatment of the whole-cell bacterial suspensions of Bordetella pertussis with 0.05% glutaraldehyde at room temperature for 10 min kills the bacteria and, except for some histamine sensitising activity, almost detoxifies the pertussis toxin. The glutaraldehyde-vaccine is of good potency with a meritorious performance in tests for abnormal toxicity in mice, the leucocytosis-promoting-factor and the mouse weight gain. Using glutaraldehyde for the inactivation of whole-cell pertussis vaccine on a commercial scale must await extensive clinical trials with vaccine lots with a good record of safety and potency in animal tests...

The authors have developed a simplified activity test for Tetanus Toxoids Adsorbed, based on the comparison of antitoxin levels in mice 4 weeks after injection of a reference toxoid and of the vaccine to be tested. Titration of tetanus antitoxin is achieved by passive agglutination of turkey RBC sensitized by means of glutaraldehyde. After preliminary experiments establishing feasibility of this method, the authors have obtained reproducible and quantitative results. They observed an increase of the immune response by a booster and an immunostimulation when pertussis component is present...

The hemagglutinating (HA) activities of purified pertussis toxin (PT) and filamentous hemagglutinin (FHA) were evaluated against unfixed and glutaraldehyde-fixed erythrocytes from ox, goose, horse, monkey, sheep, chicken, and rabbit. Both PT and FHA showed HA activities against fixed and unfixed erythrocytes from all the animals studied. The HA titers of FHA were higher than those of PT. The HA activities of FHA and PT were not destroyed completely even after heating these preparations at 56 C for 30 min. A simple test for the assay of PT in culture supernatants of Bordetella pertussis on the basis of HA activity has been described...

The immunogenicity of different types of glutaraldehyde inactivated pertussis vaccine (GIPV) preparations made by inactivation of Bordetella pertussis organisms with different concentrations of glutaraldehyde for variable periods at room temperature and conventional heat inactivated pertussis vaccine (HIPV) was evaluated with regard to production of agglutinins and neutralizing antibodies against pertussis toxin (PT). The different types of GIPV preparations had variable intracerebral mouse potency which depended upon the conditions of inactivation with glutaraldehyde...

A novel process for preparing non-pyrogenic toxoids of pertussis toxin (PT) and filamentous hemagglutinin (FHA) is described. The process consists of chromatographies on perlite then on hydroxylapatite. Purification yields for PT and FHA are 62 and 68%, respectively. The purification process takes advantage of the novel use of perlite (a filter aid) for the simultaneous purification of PT and FHA. The hydroxylapatite, in addition to removing the remaining contaminants, also concentrates the antigens. The resulting PT and FHA are approximately 95% pure, and are non-pyrogenic as judged by the rabbit pyrogen test...

Partially purified acellular pertussis vaccine was prepared from Bordetella pertussis strains 10536, 134, Tohama and 509 using simple indigenously available techniques. The Stainer-Scholte (SS) medium with methylated-beta-cyclodextrin was the most suitable for production of acellular pertussis vaccine. For preparation of the vaccine, 5 day cultures of B. pertussis grown under stationary conditions at 35 degrees C were treated twice with ammonium sulphate and prospective protective antigens were extracted. The extracts contained pertussis toxin (PT), filamentous hemagglutinin and agglutinogens...

An acellular pertussis vaccine was prepared from pertussis vaccine concentrates by eliminating the cells by centrifugation and subsequent acid precipitation. SDS-PAGE analysis indicated the presence of 17 clearly defined bands including the subunits of pertussis toxin. The acellular preparation was detoxified with glutaraldehyde and lost its undesirable effects (leukocytosis-promoting activity, histamine-sensitizing activity and decrease in weight gain) but retained a very good protective activity (PD50 approximately 2 micrograms protein/mouse)...

Two peptides, designated L and K, covering a sequence near the NH-terminal end of the S1 subunit of pertussis toxin (PT) were conjugated to the PPD (purified protein derivative) of M. tuberculosis by either glutaraldehyde (GLUT) or succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) and injected into groups of mice and guinea pigs. Initially, the effect of priming the animals with BCG vaccine and the use of aluminium hydroxide as adjuvant for the anti-peptide antibody response was studied...