Purified K‐Agglutinogen of Bordetella pertussis and Its Properties

Abstract

We attempted to purify the K-agglutinogen of Bordetella pertussis by mild procedures. From the supernatant of sonic treated cells, the K-agglutinogen was separated by a successive use of O-diethylaminoethyl (DEAE)-cellulose column chromatography, ammonium sulfate precipitation and O-carboxymethyl (CM)-cellulose column chromatography. The specific activity of purified K-agglutinogen increased 64-128 fold without damaging either the L or S antigens, and recovery was about 86%. A fraction containing mainly 5-agglutinogen, was also obtained. The K-agglutinogen, which was a simple protein with an S value of 2.5, was confirmed to be in a highly purified state by a gel-diffusion test, K-antibody production and physicochemical tests, and was also found to be free from other biologically active substances of B. pertussis: namely heat labile toxin, hemagglutinin, histamine-sensitizing factor and O-antigen. No protective activity was found in the purified K-agglutinogen or against intracerebral challenge with B. pertussis in mice.

Effects of Bordetella pertusiss Components on IgE and IgG1 Responses

“…Pertussigen was prepared from strain 3779BL 2S4 by the method of Munoz et al (33); LPS was prepared from the same strain by methods described by Nakase et al (37) and Westphal et al (53). The other antigens were prepared from the Maeno strain as follows: DNT by the method previously described (47); K-agglutinogen by a slight modification of the method of Nakase and Kasuga (38); and FHA as described by Arai and Munoz (2,33). The various activities of the purified antigens and their lack of cross contamination are given in Table 1 and Fig.…”

Antigenic Relationship Between Serotype-Specific Agglutinogen and Fimbriae of Bordetella pertussis

“…In contrast, but in agreement with previous reports (11), purified preparations of FHA similarly fixed and stained, consisted of filaments of relatively uniform length (45 nm ± 14) with a width of only 2.5 to 3.0 nm. Agglutinogen 2, prepared from a sonicate of Tohama (serotype 1,2) strain cell walls by DEAE-cellulose chromatography (8) followed by Sepharose 6B chromatography gave a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (5). This band had a subunit molecular weight of about 22,000, similar to subunits of fimbriae from other organisms (12).…”

Glycerol‐Releasing Activity of Histamine‐Sensitizing Factor of Bordetella pertussis for Rat Adipocytes In Vitro

“…The HSF was shown to be free of detectable fimbrial hemagglutinin (HA) by the method of Munoz et al (17), and of endotoxin (ET), heat-labile K agglutinogens (KA) and heatlabile dermonecrotic toxin (HLT) at a dose of I flg by the techniques described elsewhere (19)(20)(21)(22).…”

“…Partly purified preparations of HA, ET, KA, and HLT were made from strain 3779B by the previously described methods (19)(20)(21)(22).…”