Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-23T02:44:41.187Z Has data issue: false hasContentIssue false
  • English
  • Français

Use of two-colour flow cytometry to assess killing of Giardia muris trophozoites by antibody and complement

Published online by Cambridge University Press:  06 April 2009

M. F. Heyworth  and
J. Pappo
M. F. Heyworth
Affiliation:
Cell Biology Section (151E), Veterans Administration Medical Center, 4150 Clement Street, San Francisco, California 94121 and Department of Medicine, University of California, San Francisco, USA
J. Pappo
Affiliation:
Cell Biology Section (151E), Veterans Administration Medical Center, 4150 Clement Street, San Francisco, California 94121 and Department of Medicine, University of California, San Francisco, USA
Get access Rights & Permissions [Opens in a new window]

Summary

A two-colour flow cytometry technique was used to assess killing of Giardia muris trophozoites by rabbit anti-trophozoite antibodies and complement. Binding of rabbit antibody to trophozoites and killing of trophozoites were documented by flow cytometry, after incubating the organisms with fluorescein-conjugated anti-rabbit IgG and propidium iodide (PI). Percentages of PI+ (dead) trophozoites ranged from >80%, after incubation with rabbit antiserum and complement, to <30%, after incubation with complement alone. The assay technique may be applicable to studies aimed at determining whether intestinal antibodies from Giardia-infected mammals can kill Giardia trophozoites.

Keywords

Giardia murisflow cytometrytrophozoiteskillingcytotoxic antibodycomplement
Type
Research Article
Information
Parasitology , Volume 99 , Issue 2 , October 1989 , pp. 199 - 203
Copyright
Copyright © Cambridge University Press 1989

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aggarwal, A. & Nash, T. E. (1986). Lack of cellular cytotoxicity by human mononuclear cells to Giardia. Journal of Immunology 136, 3486–8.Google Scholar
Andrews, J. S. Jr., Ellner, J. J. & Stevens, D. P. (1980). Purification of Giardia muris trophozoites by using nylon fiber columns. American Journal of Tropical Medicine and Hygiene 29, 12–15.CrossRefGoogle ScholarPubMed
Belosevic, M. & Faubert, G. M. (1986). Killing of Giardia muris trophozoites in vitro by spleen, mesenteric lymph node and peritoneal cells from susceptible and resistant mice. Immunology 59, 269–75.Google Scholar
Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1984). The sensitivity of Giardia intestinalis to drugs in vitro. Journal of Antimicrobial Chemotherapy 14, 449–61CrossRefGoogle ScholarPubMed
Deguchi, M., Gillin, F. D. & Gigli, I. (1987). Mechanism of killing of Giardia lamblia trophozoites by complement. Journal of Clinical Investigation 79, 1296–302.CrossRefGoogle ScholarPubMed
Farthing, M. J. G. & Inge, P. M. G. (1986). Antigiardial activity of the bile salt-like antibiotic sodium fusidate. Journal of Antimicrobial Chemotherapy 17, 165–71.Google Scholar
Gillin, F. D. & Diamond, L. S. (1981). Inhibition of clonal growth of Giardia lamblia and Entamoeba histolytica by metronidazole, quinacrine, and other antimicrobial agents. Journal of Antimicrobial Chemotherapy 8, 305–16.CrossRefGoogle ScholarPubMed
Heyworth, M. F. (1986). Antibody response to Giardia muris trophozoites in mouse intestine. Infection and Immunity 52, 568–71.Google Scholar
Heyworth, M. F. (1988). Immunology of Giardia infections. In Advances in Giardia Research (ed. Wallis, P. M. & Hammond, B. R.), pp. 45–8. Calgary, Canada: University of Calgary Press.Google Scholar
Heyworth, M. F., Kung, J. E. & Caplin, A. B. (1988). Enzyme-linked immunosorbent assay for Giardia-specific IgA in mouse intestinal secretions. Parasite Immunology 10, 713–17.Google Scholar
Heyworth, M. F. & Pappo, J. (1987). Use of fluorescent dyes to assess viability of Giardia muris trophozoites (Abstract). Gastroenterology 92, 1435.Google Scholar
Hill, D. R., Burge, J. J. & Pearson, R. D. (1984). Susceptibility of Giardia lamblia trophozoites to the lethal effect of human serum. Journal of Immunology 132, 2046–52.CrossRefGoogle Scholar
Jackson, P. R., Pappas, M. G. & Hansen, B. D. (1985). Fluorogenic substrate detection of viable intracellular and extracellular pathogenic protozoa. Science 227, 435–8.CrossRefGoogle ScholarPubMed
Kanwar, S. S., Ganguly, N. K., Walia, B. N. S. & Mahajan, R. C. (1986). Direct and antibody dependent cell mediated cytotoxicity against Giardia lamblia by splenic and intestinal lymphoid cells in mice. Gut 27, 73–7.Google Scholar
Kaplan, B. S., Uni, S., Aikawa, M. & Mahmoud, A. A. F. (1985). Effector mechanism of host resistance in murine giardiasis: specific IgG and IgA cell-mediated toxicity. Journal of Immunology 134, 1975–81.CrossRefGoogle ScholarPubMed
Rohrer, L., Winterhalter, K. H., Eckert, J. & Köhler, P. (1986). Killing of Giardia lamblia by human milk is mediated by unsaturated fatty acids. Antimicrobial Agents and Chemotherapy 30, 254–7.Google Scholar
Schaefer, F. W. III, Rice, E. W. & Hoff, J. C. (1984). Factors promoting in vitro excystation of Giardia muris cysts. Transactions of the Royal Society of Tropical Medicine and Hygiene 78, 795–800.CrossRefGoogle ScholarPubMed
Schupp, D. G. & Erlandsen, S. L. (1987). A new method to determine Giardia cyst viability: correlation of fluorescein diacetate and propidium iodide staining with animal infectivity. Applied and Environmental Microbiology 53, 704–7.Google Scholar
Smith, P. D., Elson, C. O., Keister, D. B. & Nash, T. E. (1982). Human host response to Giardia lamblia. I. Spontaneous killing by mononuclear leukocytes in vitro. Journal of Immunology 128, 1372–6.Google Scholar
Snider, D. P., Gordon, J., Mcdermott, M. R. & Underdown, B. J. (1985). Chronic Giardia muris infection in anti-IgM-treated mice. I. Analysis of immunoglobulin and parasite-specific antibody in normal and immunoglobulin-deficient animals. Journal of Immunology 134, 4153–62.CrossRefGoogle ScholarPubMed