Characterization of an RTX toxin from enterohemorrhagic Escherichia coli O157: H7

Margaret Bauer, Rodney A. Welch

Research output: Contribution to journalArticle

119 Citations (Scopus)

Abstract

A hemolytic determinant of enterohemorrhagic Escherichia coli O157:H7 is encoded on a 90-kbp plasmid (pO157). This enterohemorrhagic E. coli toxin (Ehx) is a newly described RTX cytotoxin. The prototype RTX toxin is the E. coli hemolysin (Hly) associated with extraintestinal E. coli infections. We expressed Ehx from E. coli K-12 strains harboring either pSK3, a pO157 derivative marked with Tn801 unlinked to Ehx, or a recombinant plasmid containing an 11.9-kbp subclone (pEO40) of pSK3. The Ehx activities and antibody reactivities were compared with those of Hly. Little Ehx was secreted extracellularly from the strain harboring pSK3; however, when the Hly transport genes hlyBD were supplied in trans, both intracellular and extracellular levels of Ehx were enhanced more than 15-fold. The strain harboring pEO40 secreted at least 140-fold more Ehx than did the strain harboring pSK3, and neither intracellular nor extracellular levels were significantly enhanced by the addition of hlyBD in trans. Polyclonal anti-HlyA antiserum and several anti-HlyA monoclonal antibodies, including the monoclonal antibody A10, which is panreactive for nearly all RTX toxins, reacted with EhxA antigen by immunoblot analysis. In hemolysis and 51Cr release assays, Ehx demonstrated similar efficiencies in lysis of BL-3 cells (cells from a bovine lymphoma cell line) and sheep and human erythrocytes. Surprisingly, it demonstrated very little activity against two human lymphoma cell lines. In contrast, Hly lysed all five cell types tested, each to a greater extent than that demonstrated by comparable amounts of Ehx. As with other RTX toxins, Ehx activity was calcium dependent and heat labile.

Original languageEnglish (US)
Pages (from-to)167-175
Number of pages9
JournalInfection and Immunity
Volume64
Issue number1
StatePublished - 1996
Externally publishedYes

Fingerprint

Enterohemorrhagic Escherichia coli
Hemolysin Proteins
Escherichia coli O157
Plasmids
Monoclonal Antibodies
Enzootic Bovine Leukosis
Cell Line
Escherichia coli Infections
antineoplaston A10
Cytotoxins
Hemolysis
Immune Sera
Lymphoma
Sheep
Hot Temperature
Erythrocytes
Escherichia coli
Calcium
Antigens
Antibodies

ASJC Scopus subject areas

  • Immunology

Cite this

Characterization of an RTX toxin from enterohemorrhagic Escherichia coli O157 : H7. / Bauer, Margaret; Welch, Rodney A.

In: Infection and Immunity, Vol. 64, No. 1, 1996, p. 167-175.

Research output: Contribution to journalArticle

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