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Infect Immun. 1981 July; 33(1): 59-66

Increased virulence of Neisseria meningitidis after in vitro iron-limited growth at low pH.

ABSTRACT

At low pH (6.6) and under conditions of iron limitation, Neisseria meningitidis group B (strain SD1C) exhibited an atypical outer membrane protein profile and an increased relative virulence for the mouse. Cells grown in a buffered medium were effectively deprived of iron by the addition of ethylenediamine-diorthohydroxyphenylacetate. The pH of the medium selected for characteristic colonial morphologies: type M3 predominated at pH 6.6, and type M5 predominated at pH 7.7. A mixed population of M1, M3, and M5 colonies was observed at pH 7.2. Isolated outer membrane proteins were analyzed by sodium dodecyl 99 99 sulfate-polyacrylamide gel electrophoresis, and surface exposed proteins were labeled by the [125I]lactoperoxidase method and subsequently identified by autoradiography. Cells grown at pH 6.6 elaborated a major outer membrane protein (protein III; molecular weight, 69,000), which was also present in the outer membrane of iron-limited cells grown at pH 7.2. At pH 7.2 in an iron-sufficient medium, protein III was present only in small quantities in sodium dodecyl sulfate-polyacrylamide gel was present only in small quantities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels. A study of the relative virulence (50% lethal dose) of the meningococcus for C57/BL mice revealed that iron-limited cells grown at low pH had an increased relative virulence 1,200-fold (50% lethal dose, 4.0 CFU) greater than that of cells grown in the same medium but at pH 7.2 and with sufficient iron. These studies indicate that pH and iron can be important factors in the determination of meningococcal virulence.

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