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Cancer Chemotherapy and Pharmacology

, Volume 36, Issue 2, pp 115–120 | Cite as

Preclinical pharmacology of cholera toxin

  • Joel M. Reid
  • John W. Benson
  • Jean Viallet
  • Matthew M. Ames
Original Article Holera Toxin, Immunoassay, Pharmacokinetics

Abstract

Cholera toxin was selected for pharmacologic evaluation by the National Cancer Institute on the basis of antiproliferative activity against small-cell and nonsmall-cell lung-cancer cell lines. A feature common to the sensitive cell lines was abundant expression of GM1 ganglioside, the cellular receptor for cholera toxin. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to quantitate cholera toxin in biological fluids. A sigmoidal relationship was observed between the cholera toxin plasma concentration and the absorbance at 490 nm (OD490) of the product of horseradish peroxidase-catalyzed oxidation ofo-phenylenediamine over the range of 6.25–1,600 ng/ml. Logit transformation of the OD490 data was linear over the entire concentration range and assay variability was less than 25%. Cholera toxin was stable in murine and human whole blood and plasma. Following i.v. administration of 1,500 μg/kg to male CD2F1 mice, cholera toxin plasma elimination was described by a two-compartment open model. The half-lives (t1/2α,t1/2β), plasma clearance, and steady-state volume of distribution were 0.7 min, 49 min, 24 ml min−1 kg−1 912 ml/kg, respectively. Cholera toxin was not detected in plasma following an s.c. dose of 1,500 μg/kg. Urinary recovery following intravenous drug administration was less than 0.1%.

Key words

Cholera toxin Immunoassay Pharmacokinetics 

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References

  1. 1.
    Beutin L, Bode L, Richter T, Peltre G, Stephan R (1984) Rapid visual detection ofEscherichia coli andVibrio cholerae heatlabile enterotoxins by nitrocellulose enzyme-linked immunosorbent assay. J Clin Microbiol 19:371Google Scholar
  2. 2.
    Buckler JMH (1970) A comparison of the effects of human serum in two double antibody radioimmunoassay systems for the estimation of luteinizing hormone. In: Kirkham KE, Hunter WM (eds) Radioimmunoassay methods, European workshop, September 15–17, Edinburgh. Williams & Wilkins, Baltimore, p 273Google Scholar
  3. 3.
    Chisari FV, Northrop RS (1974) Pathophysiologic effects of lethal and immunoregulatory doses of cholera enterotoxin in the mouse. J Immunol 113:740Google Scholar
  4. 4.
    Cho-Chung Y, Clair T, Shepheard C, Berghoffer B (1983) Arrest of hormone-dependent mammary cancer growth in vivo and in vitro by cholear toxin. Cancer Res 43:1473Google Scholar
  5. 5.
    Court G, Hurn BAL (1970) An effect of plasma and other macromolecular solution in double antibody radioimmunoassary systems. In: Kirkham KE, Hunter WM (eds) Radioimmunosaasy methods, european workshop, September 15–17, Edinburgh. Williams & Wilkins, Baltimore, p 273Google Scholar
  6. 6.
    Cuatrecasas P (1973) Interaction ofVibrio cholerae entertoxin with cell membrances. Biochemistry 12:3547Google Scholar
  7. 7.
    De Lean AP, Munson PJ, Rodbard D (1978) Simultaneous analysis of families of sigmoidal curves: application to bioassay, radioligand assay, and physiological dose-response curves. Endocrinol Metab Gastrointest Physiol 235:E97Google Scholar
  8. 8.
    Greenberg HB, Sack DA, Rodriguez W, Sack RB, Wyatt RG, Kalica AR, Horswood RL, Chanock RM, Kapikian AZ (1977) Microtiter solid-phase radioimmunoassay for detection ofEscherichia coli heat-labile enterotixin. Infect Immun 17:541Google Scholar
  9. 9.
    Harlow E, Lane D (1988) Immunoassays. In: Harlow E, Lane D (eds) Antibodies: a laboratory manual Cold Spring Harbor Press, Cold Spring Harbor, p 553Google Scholar
  10. 10.
    Heitzmann H, Richards FM (1974) Use of the avidin-biotin complex for specific staining of biological membranes in electron microscopy. PNAS 71:3537Google Scholar
  11. 11.
    Holmgren J (1981) Actions of cholera toxin and prevention and treatment of cholera. Nature 292:413Google Scholar
  12. 12.
    Holmgren J, Lange S, Lindholm L, Lonnroth I (1977) In vivo modulation of intracellular cAMP and cell growth of a lymphatic tumour in mice by cholera toxin. Exp Cell Res 108:31Google Scholar
  13. 13.
    Karpinski KF, Hayward S, Tryphonas H (1987) Statistical considerations in the quantitation of serum immunoglobulin levels using the enzyme-linked immunosorbent assay (ELISA). J Immunol Methods 103:189Google Scholar
  14. 14.
    Kaur G, Viallet J, Laborda J, Blair O, Gazdar AF, Minna JD, Sausville EA (1992) Growth inhibition by cholera toxin of human lung carcinoma cell lines: correlation with GM1 ganglioside expression. Cancer Res 52:3340Google Scholar
  15. 15.
    Knauf MJ, Bell DP, Hirtzer P, Luo Z-P, Katre NV (1988) Relationship of effective molecular size to systemic clearance in rats of recombinant interleukin-2 chemically modified with water-soluble polymers. J Biol Chem 263:15064Google Scholar
  16. 16.
    Liu CT, Galloway EJ, Loizeaux PS (1980) Cardiohepatic and gross pathological changes in rhesus monkeys after intravenous injection of purified cholera enterotoxin. Toxicon 18:309Google Scholar
  17. 17.
    Maack T, Johnson V, Kau ST, Figueiredo J, Sigulem D (1979) Renal filtration, transport, and metabolism of low-molecularweight proteins: a review: Kidney Int 16:251Google Scholar
  18. 18.
    Marks DL, Gores GJ, LaRusso NF (1991) Hepatic processing of the proteins and peptides by the perfused liver. In: Ballet F, Thurman RG (eds) Research in perfused liver. INSERM/John Libbey, London, P 209Google Scholar
  19. 19.
    Moss J, Vaughan M (1988) ADP-ribosylation of guanyl nucleotide-binding regulatory proteins by bacterial toxins. Adv Enzymol 61:303Google Scholar
  20. 20.
    Oku Y, Uesaka Y, Hirayama T, Takeda Y (1988) Development of a highly sensitive bead-ELISA to detect bacterial protein toxins. Microbiol Immunol 32:807Google Scholar
  21. 21.
    Pierce NF, Graybill R, Kaplan MM, Bouwman DL (1972) Systemic effects of parenteral cholera enterotoxin in dogs. J Lab Clin Med 79:145Google Scholar
  22. 22.
    Pimplikar SW, Simons K (1993) Regulation of apical transport in epithelial cells by Gs glass of heteotrimeric G proteins Nature 362:456Google Scholar
  23. 23.
    Svennerholm A-M, Holmgren J (1978) Identification ofEscherichia coli heat-labile enterotoxin by means of a ganglioside immunosorbent assay (GM1-ELISA) procedure. Curr Microbiol 1:19Google Scholar
  24. 24.
    Svennerholm A-M, Wiklund G (1983) Rapid GM1-enzyme-linked immunosorbent assay with visual reading for identification ofEscherichia coli heat-labile enterotoxin. J Clin Microbiol 17:596Google Scholar
  25. 25.
    Tijssen P (1985) Processing of data and reporting of results of enzyme immunoassays. In: Laboratory techniques in biochemistry and molecular biology. Practice and theory of enyzme immunoassays. Elsevier, Amsterdam, p 385Google Scholar
  26. 26.
    Tsuru S, Taniguchi M, Shinomiya N, Fujisawa H, Zinnaka Y, Nomoto K (1987) Cholera toxin-induced tolerance to allografts in mice. Immunology 61:77Google Scholar
  27. 27.
    Viallet J, Sharoni Y, Frucht H, Jensen RT, Minna JD, Sausville EA (1990) Cholera toxin inhibits signal transduction by several mitogens and the in vitro growth of human small-cell lung cancer. J Clin Invest 86:1904Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Joel M. Reid
    • 1
  • John W. Benson
    • 1
  • Jean Viallet
    • 2
  • Matthew M. Ames
    • 1
  1. 1.Department of Oncology, Division of Developmental Oncology ResearchMayo Clinic and FoundationRochesterUSA
  2. 2.Department of OncologyMcGill University and Montreal General HospitalMontrealCanada

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