[Biochemical and serological investigations of flagellae of Campylobacter fetus (author's transl)].
Flagellae of Campylobacter fetus group O, types 1, 2 and 7 were prepared. First they were separated from cell bodies using an ultramix. The suspension was then centrifuged for 20 mins. at 10000 rpm and the supernatant frozen at -40 degrees C. This is a simple method for the enrichment of preparations of flagellae, as they become tangled up and accumulate in the inferior third part of the frozen liquid. The physicochemical basis of this phenomenon was discussed. After thawing and spinning for 20 mins. at 5000 rpm, the sediment was suspended in 0.9% of NaC1. The purity of the preparation was checked by electron microscopy. Antibodies to this antigen showed no cross-reaction with O-Antigen, when tested by tube agglutination. The amino acid composition of flagellae from different O-antigen serotypes was different (see Tab. 1). Cysteine could not be detected and proline only in traces. After breakdown with urea followed by gel filtration on Sephadex G 200, breakdown products of diminishing molecular size were obtained (see Fig. 2). Discelectrophoresis after ultrasonic gave 8 zones (see Fig. 3). Irrespective of serotype, thin-layer chromatography of trypsin-hydrolysed flagellae always showed 9 ninhydrin-positive spots (see Fig. 1). Only breakdown products of ultrasonic reacted with antibody. After absorption of flagellar antibody with heterologous antigen, agglutination only occurred with homologous antigen (tab. 2-5). This showed that there were different flagellar antigens. Further experiments using immunoprecipitation demonstrated two common antigenic components, a and c, and a partially common antigenic factor bb (Fig. 4), and was the basis for a classification by three groups. The three antigenic components could be separated by gel electrophoresis and detected by immunoprecipitation (Figs. 5,6).
D000372:Agglutination Tests; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000942:Antigens, Bacterial; D002167:Campylobacter / Q000276:immunology*; D002168:Campylobacter fetus / Q000276:immunology*; D002458:Cell Fractionation; D005407:Flagella / Q000032:analysis / Q000276:immunology*; D011233:Precipitin Tests; D012703:Serotyping
D000596:Amino Acids; D000942:Antigens, Bacterial
Biochemische und serologische Untersuchungen an Geisseln von Campylobacter fetus