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. 1991 Jan;35(1):29–35. doi: 10.1128/aac.35.1.29

In vitro model of attachment of Giardia intestinalis trophozoites to IEC-6 cells, an intestinal cell line.

R E McCabe 1, G S Yu 1, C Conteas 1, P R Morrill 1, B McMorrow 1
PMCID: PMC244937  PMID: 1901700

Abstract

Attachment of giardias to intestinal cells has been difficult to study because of a lack of a convenient in vitro model. We developed an assay for attachment of radiolabeled trophozoites to IEC-6 cells that can be done in microtiter trays. Attachment was confirmed by scanning and transmission electron microscopy. Trophozoites remained attached to the IEC-6 cells for 24 h with little evidence of damage to the IEC-6 cells. Preincubation of trophozoites with cytochalasins A, B, and D reduced attachment to approximately 20% of that of controls, whereas colchicine had no effect. Chelation of divalent cations with EDTA and EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid] reduced attachment to 24 and 26% of control values, respectively, and incubation at 4 degrees C reduced attachment to 7% of the value for controls incubated at 37 degrees C. Glutaraldehyde fixation of trophozoites or IEC-6 cells resulted in significantly diminished attachment to the live substrate (17 and 40% of control values, respectively). Coincubation of IEC-6 cells and trophozoites on a rotary shaker resulted in detachment of 40% of trophozoites, but EDTA, EGTA, glutaraldehyde fixation of trophozoites, and low temperature diminished attachment markedly and significantly. Similar results were obtained in selected experiments with three strains of giardia.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bouhours J. F., Glickman R. M. Rat intestinal glycolipids. II. Distribution and biosynthesis of glycolipids and ceramide in villus and crypt cells. Biochim Biophys Acta. 1976 Jul 20;441(1):123–133. doi: 10.1016/0005-2760(76)90287-3. [DOI] [PubMed] [Google Scholar]
  2. Chávez B., Knaippe F., Gonzalez-Mariscal L., Martínez-Palomo A. Giardia lamblia: electrophysiology and ultrastructure of cytopathology in cultured epithelial cells. Exp Parasitol. 1986 Jun;61(3):379–389. doi: 10.1016/0014-4894(86)90194-3. [DOI] [PubMed] [Google Scholar]
  3. Conteas C. N., Majumdar A. P. The effects of gastrin, epidermal growth factor, and somatostatin on DNA synthesis in a small intestinal crypt cell line (IEC-6). Proc Soc Exp Biol Med. 1987 Mar;184(3):307–311. doi: 10.3181/00379727-184-42484. [DOI] [PubMed] [Google Scholar]
  4. Farthing M. J., Pereira M. E., Keusch G. T. Description and characterization of a surface lectin from Giardia lamblia. Infect Immun. 1986 Feb;51(2):661–667. doi: 10.1128/iai.51.2.661-667.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Farthing M. J., Pereira M. E., Keusch G. T. Giardia lamblia: evaluation of roller bottle cultivation. Exp Parasitol. 1982 Dec;54(3):410–415. doi: 10.1016/0014-4894(82)90050-9. [DOI] [PubMed] [Google Scholar]
  6. Feely D. E., Erlandsen S. L. Effect of cytochalasin-B, low Ca++ concentration, iodoacetic acid, and quinacrine-HCl on the attachment of Giardia trophozoites in vitro. J Parasitol. 1982 Oct;68(5):869–873. [PubMed] [Google Scholar]
  7. Feely D. E., Schollmeyer J. V., Erlandsen S. L. Giardia spp.: distribution of contractile proteins in the attachment organelle. Exp Parasitol. 1982 Feb;53(1):145–154. doi: 10.1016/0014-4894(82)90100-x. [DOI] [PubMed] [Google Scholar]
  8. Friend D. S. The fine structure of Giardia muris. J Cell Biol. 1966 May;29(2):317–332. doi: 10.1083/jcb.29.2.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gillin F. D., Reiner D. S. Attachment of the flagellate Giardia lamblia: role of reducing agents, serum, temperature, and ionic composition. Mol Cell Biol. 1982 Apr;2(4):369–377. doi: 10.1128/mcb.2.4.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Godman G. C., Miranda A. F. Cellular contractility and the visible effects of cytochalasin. Front Biol. 1978;46:277–429. [PubMed] [Google Scholar]
  11. Holberton D. V. Attachment of Giardia-a hydrodynamic model based on flagellar activity. J Exp Biol. 1974 Feb;60(1):207–221. doi: 10.1242/jeb.60.1.207. [DOI] [PubMed] [Google Scholar]
  12. Holberton D. V. Fine structure of the ventral disk apparatus and the mechanism of attachment in the flagellate Giardia muris. J Cell Sci. 1973 Jul;13(1):11–41. doi: 10.1242/jcs.13.1.11. [DOI] [PubMed] [Google Scholar]
  13. Inge P. M., Edson C. M., Farthing M. J. Attachment of Giardia lamblia to rat intestinal epithelial cells. Gut. 1988 Jun;29(6):795–801. doi: 10.1136/gut.29.6.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Keister D. B. Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg. 1983;77(4):487–488. doi: 10.1016/0035-9203(83)90120-7. [DOI] [PubMed] [Google Scholar]
  15. Kobiler D., Mirelman D. Adhesion of Entamoeba histolytica trophozoites to monolayers of human cells. J Infect Dis. 1981 Dec;144(6):539–546. doi: 10.1093/infdis/144.6.539. [DOI] [PubMed] [Google Scholar]
  16. Kurokowa M., Lynch K., Podolsky D. K. Effects of growth factors on an intestinal epithelial cell line: transforming growth factor beta inhibits proliferation and stimulates differentiation. Biochem Biophys Res Commun. 1987 Feb 13;142(3):775–782. doi: 10.1016/0006-291x(87)91481-1. [DOI] [PubMed] [Google Scholar]
  17. Lev B., Ward H., Keusch G. T., Pereira M. E. Lectin activation in Giardia lamblia by host protease: a novel host-parasite interaction. Science. 1986 Apr 4;232(4746):71–73. doi: 10.1126/science.3513312. [DOI] [PubMed] [Google Scholar]
  18. Owen R. L., Nemanic P. C., Stevens D. P. Ultrastructural observations on giardiasis in a murine model. I. Intestinal distribution, attachment, and relationship to the immune system of Giardia muris. Gastroenterology. 1979 Apr;76(4):757–769. [PubMed] [Google Scholar]
  19. Quaroni A. Development of fetal rat intestine in organ and monolayer culture. J Cell Biol. 1985 May;100(5):1611–1622. doi: 10.1083/jcb.100.5.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rădulescu S., Rău C., Petraşincu D., Gaicu N., Meyer E. A. Behaviour and cytopathogenicity of Giardia lamblia in cell cultures. Arch Roum Pathol Exp Microbiol. 1980 Apr-Jun;39(2):163–170. [PubMed] [Google Scholar]
  21. Zenian A., Gillin F. D. Interactions of Giardia lamblia with human intestinal mucus: enhancement of trophozoite attachment to glass. J Protozool. 1985 Nov;32(4):664–668. doi: 10.1111/j.1550-7408.1985.tb03098.x. [DOI] [PubMed] [Google Scholar]

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