Isolation and Characterization of Isogenic Pairs of Domed Hemolytic and Flat Nonhemolytic Colony Types of Bordetella pertussis
- 1 March 1982
- journal article
- research article
- Published by American Society for Microbiology in Infection and Immunity
- Vol. 35 (3), 840-851
- https://doi.org/10.1128/iai.35.3.840-851.1982
Abstract
Four diferent serotype strains of B. pertussis (3779BL2S4, Tohama I, 353/Z and 2753) were plated on Bordet-Gengou agar; they grew as domed, hemolytic (D+H+) wild-type colonies. Cloned D+H+ colony types of all 4 strains were passed onto modified Stainer-Scholte medium solidified with 1% Noble Agar. Colonies were selected from Stainer-Scholte agar; these subsequently grew as flat, nonhemolytic (D-H-) colonies when transferred back onto Bordet-Gengou agar. The frequency of D-H- organisms within a population of cloned D+H+ was determined to be between 5 .times. 10-5-5 .times. 10-6. The D-H- colony types maintained their flat, nonhemolytic characteristics for over 80 single-colony passages on Bordet-Gengou agar. The isogenic pairs of D+H+ and D-H- colony types from the 4 strains were compared for hemagglutination titer, lymphocytosis-promoting activity, adenylate cyclase activity and presence of agglutinogens by agglutination. In all cases, the D-H- colony types showed reduced activities or amounts of antigen compared with their D+H+ parents. Freely diffusible antigens were markedly different between the 2 phenotypes as noted by double diffusion of antisera added to plates on which colonies of the variants were growing. Antigens solubilized from the 2 colony types by Triton X-100 were also markedly different as judged by radial immunodiffusion with antifimbrial hemagglutinin, antilymphocytosis-promoting factor and anti-353/Z adsorbed with autoclaved 353/Z. Autoradiographs of 125I-surface-labeled whole cells separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed unique banding patterns for each colony type. Since all organisms, regardless of colony type, were grown on Bordet-Gengou agar, the differences reported could not be due to medium composition. Differences between phenotypes were also independent of passage number on Bordet-Gengou agar. By analogy to previous studies, the D-H- organisms appear to fulfill the criteria for phase III or phase IV in the system of Leslie and Gardner or phase III in the system of Kasuga.This publication has 27 references indexed in Scilit:
- Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphenylglycolurilBiochemical and Biophysical Research Communications, 1978
- Selection from Gonococci Grown in vitro of a Colony Type with some Virulence Properties of Organisms Adapted in vivoJournal of General Microbiology, 1977
- The importance of agglutinin production in mice in the determination of the definitive serotype ofBordetella pertussisEpidemiology and Infection, 1976
- Cell-Envelope Proteins of Bordetella PertussisJournal of Medical Microbiology, 1975
- A reexamination of phase IV Bordetella pertussisCanadian Journal of Microbiology, 1972
- A Simple Chemically Defined Medium for the Production of Phase I Bordetella pertussisJournal of General Microbiology, 1970
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Pitfalls in the Preparation of Monotypic Agglutinating Antisera for Bordetella PertussisJournal of Medical Microbiology, 1968
- PRESERVATION OF LIVING CELLS BY FREEZE‐DRYINGAnnals of the New York Academy of Sciences, 1960
- The Growth of Bordetella pertussis: a ReviewJournal of General Microbiology, 1957