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Applied and Environmental Microbiology, October 2003, p. 5907-5913, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5907-5913.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Effects of Iron Limitation on Adherence and Cell Surface Carbohydrates of Corynebacterium diphtheriae Strains

Lílian de Oliveira Moreira,1,2 Arnaldo Feitosa Braga Andrade,1 Márcio Damasceno Vale,1 Sônia Maria Silva Souza,1,3 Raphael Hirata Jr.,1 Lídia Maria Oliveira Buarque Asad,3 Nasser Ribeiro Asad,3 Luiz Henrique Monteiro-Leal,3 José Osvaldo Previato,2 and Ana Luiza Mattos-Guaraldi1*

Faculdade de Ciências Médicas,1 Instituto de Biologia Prof. Roberto AlcÂntara Gomes, Universidade do Estado do Rio de Janeiro,3 Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil2

Received 2 December 2002/ Accepted 18 July 2003

Iron limitation may cause bacterial pathogens to grow more slowly; however, it may also stimulate these microorganisms to produce greater tissue damage, given that many virulence factors are controlled by the iron supply in the environment. The present study investigated the influence of low iron availability on the expression of proteins and surface sugar residues of two toxigenic strains of Corynebacterium diphtheriae subsp. mitis and evaluated their adherence to human group B erythrocytes and HEp-2 cells. A comparison was made between bacteria grown in (i) Trypticase soy broth (TSB), (ii) TSB treated with dipyridyl to deplete free iron, and (iii) TSB enriched with FeCl3. The effects of iron concentration on adhesive properties were different for strains 241 and CDC-E8392, of the sucrose-fermenting and non-sucrose-fermenting biotypes, respectively. Iron-limited conditions enhanced interaction of strain 241 with erythrocytes and HEp-2 cells. Inhibition assays suggested the involvement of nonfimbrial protein combination 67-72p on hemagglutination of diphtheria bacilli grown under iron-limited conditions. Conversely, iron limitation inhibited adherence to glass and expression of electron-dense material on the bacterial surface. Lectin binding assays demonstrated a reduction in the number of sialic acid residues and an increase in D-mannose and D-galactose residues on the surfaces of both strains. Thus, iron exerts a regulatory role on adhesive properties of diphtheria bacilli, and low iron availability modulates the expression of C. diphtheriae surface carbohydrate moieties. The significant changes in the degree of lectin binding specific for D-mannose, D-galactose and sialic acid residues may have an effect on binding of host cells. The expression of dissimilar microbial virulence determinants may be coordinately controlled by common regulatory systems. For C. diphtheriae, the present results imply regulation of adherence and slime production as part of a global response to iron-limited environmental conditions that includes derepression of genes for the synthesis of cytotoxin and siderophores and for transport of the Fe(III)-siderophore complexes.

* Corresponding author. Mailing address: Disciplina de Microbiologia e Imunologia, Faculdade de Ciências Médicas, Universidade do Estado do Rio de Janeiro, Av. 28 de Setembro, 87 Fundos, no. 3 andar, Vila Isabel, RJ CEP 20 551-030, Brazil. Phone: 55 (21) 2587-6380. Fax: 55 (21) 2587-6476. E-mail: guaraldi{at}uerj.br.

Applied and Environmental Microbiology, October 2003, p. 5907-5913, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5907-5913.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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