Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology: a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli

R. L. Guerrant, L. L. Brunton, T. C. Schnaitman, L. I. Rebhun, A. G. Gilman

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Abstract

The major limitation of the clinical importance of enterotoxigenic E.coli in diarrheal illness is the lack of a simple rapid assay for the enterotoxin produced by certain E.coli. On the basis of the activation of adenylate cyclase by heat labile enterotoxin of E.coli (LT) and by cholera toxin (CT) in intestinal and other tissues, cultured Chinese hamster ovary (CHO) cells with known morphological responses to dibutyryl cyclic adenosine 5' monophosphate (AMP) were exposed to these enterotoxins. Crude culture filtrates of LT producing E.coli and CT stimulated cyclic AMP accumulation and cell elongation in CHO cells. The similarity of time course, concentration dependence, and potentiation by phosphodiesterase inhibitors suggested cyclic AMP mediation of the morphological change. Heat inactivated CT and LT in this system. Choleragenoid inhibited CT; antiserum against CT inhibited both enterotoxin effects. In contrast to culture filtrates of 16 strains of E.coli known to produce LT, culture filtrates from 13 E.coli that do not produce LT did not alter CHO cell morphology. The morphological change is a simple, specific assay for these enterotoxins and detect 3 x 10 -17 mol of CT or a 1:250 dilution of crude culture filtrate of LT producing E.coli 334.

Original languageEnglish (US)
Pages (from-to)320-327
Number of pages8
JournalInfection and Immunity
Volume10
Issue number2
StatePublished - 1974

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Vibrio cholerae
Cholera Toxin
Enterotoxins
Cricetulus
Cyclic AMP
Ovary
Escherichia coli
Enterotoxigenic Escherichia coli
Phosphodiesterase Inhibitors
In Vitro Techniques
Adenylyl Cyclases
Immune Sera
Hot Temperature

ASJC Scopus subject areas

  • Immunology

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Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology : a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli. / Guerrant, R. L.; Brunton, L. L.; Schnaitman, T. C.; Rebhun, L. I.; Gilman, A. G.

In: Infection and Immunity, Vol. 10, No. 2, 1974, p. 320-327.

Research output: Contribution to journalArticle

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