Abstract
In a study to investigate early interactions of Entamoeba histolytica with epithelial cell monolayers, we found that a monoclonal antibody (MAb), CD6, against an ameba surface antigen recognized the lateral surface of epithelial cells after coculture with trophozoites. Display of the CD6 antigen on the epithelial cells necessitated contact with active trophozoites. It was found neither at 4 degrees C, nor with prefixed trophozoites, nor with trophozoite-conditioned media, nor when a filter prevented direct contact. Monolayers exposed to amebic sonicates or detergent lysates showed random immunostaining. Access to the antigenic site was limited, as immunostaining occurred predominantly with subconfluent monolayers. CD6 epithelial cell binding was first observed after 5 min of coculture; trophozoite-mediated target cell lysis was not detected until 15 min following coculture. Epithelial cell immunostaining occurred with some other ameba-specific antibodies but not with MAbs raised against the 170-kDa subunit of the galactose-N-acetylgalactosamine (Gal/GalNAc)-specific lectin. The CD6 MAb as well as some other ameba-specific antibodies immunoprecipitated from trophozoite lysates the same bands as the MAbs against the cysteine-rich domain of the 170-kDa subunit of the Gal/GalNAc-specific lectin. Why the latter MAbs failed to stain epithelial cells in the vicinity of attached trophozoites is presently unknown. We concluded that E. histolytica trophozoites transferred the intact amebic Gal/GalNAc-specific lectin or a portion of it to the lateral surface of epithelial cells. This juxtacrine transfer preceded killing of target cells.
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- Avila E. E., Calderon J. Entamoeba histolytica trophozoites: a surface-associated cysteine protease. Exp Parasitol. 1993 May;76(3):232–241. doi: 10.1006/expr.1993.1028. [DOI] [PubMed] [Google Scholar]
- Azzarone B., Chaponnier C., Krief P., Mareel M., Suarez H., Macieira-Coelho A. Human fibroblasts from cancer patients: lifespan and transformed phenotype in vitro and role of mesenchyme in vivo. Mutat Res. 1988 Jun;199(2):313–325. doi: 10.1016/0027-5107(88)90211-4. [DOI] [PubMed] [Google Scholar]
- Bailey G. B., Gilmour J. R., McCoomer N. E. Roles of target cell membrane carbohydrate and lipid in Entamoeba histolytica interaction with mammalian cells. Infect Immun. 1990 Jul;58(7):2389–2391. doi: 10.1128/iai.58.7.2389-2391.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burchard G. D., Prange G., Mirelman D. Interaction between trophozoites of Entamoeba histolytica and the human intestinal cell line HT-29 in the presence or absence of leukocytes. Parasitol Res. 1993;79(2):140–145. doi: 10.1007/BF00932260. [DOI] [PubMed] [Google Scholar]
- Diamond L. S., Harlow D. R., Cunnick C. C. A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg. 1978;72(4):431–432. doi: 10.1016/0035-9203(78)90144-x. [DOI] [PubMed] [Google Scholar]
- Flores B. M., Reed S. L., Ravdin J. I., Torian B. E. Serologic reactivity to purified recombinant and native 29-kilodalton peripheral membrane protein of pathogenic Entamoeba histolytica. J Clin Microbiol. 1993 Jun;31(6):1403–1407. doi: 10.1128/jcm.31.6.1403-1407.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Griffiths J. K., Moore R., Dooley S., Keusch G. T., Tzipori S. Cryptosporidium parvum infection of Caco-2 cell monolayers induces an apical monolayer defect, selectively increases transmonolayer permeability, and causes epithelial cell death. Infect Immun. 1994 Oct;62(10):4506–4514. doi: 10.1128/iai.62.10.4506-4514.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gumbiner B., Simons K. A functional assay for proteins involved in establishing an epithelial occluding barrier: identification of a uvomorulin-like polypeptide. J Cell Biol. 1986 Feb;102(2):457–468. doi: 10.1083/jcb.102.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holgate C. S., Jackson P., Cowen P. N., Bird C. C. Immunogold-silver staining: new method of immunostaining with enhanced sensitivity. J Histochem Cytochem. 1983 Jul;31(7):938–944. doi: 10.1177/31.7.6189883. [DOI] [PubMed] [Google Scholar]
- Leippe M., Tannich E., Nickel R., van der Goot G., Pattus F., Horstmann R. D., Müller-Eberhard H. J. Primary and secondary structure of the pore-forming peptide of pathogenic Entamoeba histolytica. EMBO J. 1992 Oct;11(10):3501–3506. doi: 10.1002/j.1460-2075.1992.tb05432.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leroy A., Mareel M., De Bruyne G., Bailey G., Nelis H. Metastasis of Entamoeba histolytica compared to colon cancer: one more step in invasion. Invasion Metastasis. 1994;14(1-6):177–191. [PubMed] [Google Scholar]
- Li E., Becker A., Stanley S. L., Jr Use of Chinese hamster ovary cells with altered glycosylation patterns to define the carbohydrate specificity of Entamoeba histolytica adhesion. J Exp Med. 1988 May 1;167(5):1725–1730. doi: 10.1084/jem.167.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li E., Stenson W. F., Kunz-Jenkins C., Swanson P. E., Duncan R., Stanley S. L., Jr Entamoeba histolytica interactions with polarized human intestinal Caco-2 epithelial cells. Infect Immun. 1994 Nov;62(11):5112–5119. doi: 10.1128/iai.62.11.5112-5119.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- López-Revilla R., Reyes J. L., Enríquez-Rincón F., Basurto M. A. Immediate deterioration of rabbit colon preparations by Entamoeba histolytica trophozoite lysates. Parasitol Res. 1992;78(3):260–262. doi: 10.1007/BF00931737. [DOI] [PubMed] [Google Scholar]
- Mann B. J., Chung C. Y., Dodson J. M., Ashley L. S., Braga L. L., Snodgrass T. L. Neutralizing monoclonal antibody epitopes of the Entamoeba histolytica galactose adhesin map to the cysteine-rich extracellular domain of the 170-kilodalton subunit. Infect Immun. 1993 May;61(5):1772–1778. doi: 10.1128/iai.61.5.1772-1778.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martínez-Palomo A., González-Robles A., Chávez B., Orozco E., Fernández-Castelo S., Cervantes A. Structural bases of the cytolytic mechanisms of Entamoeba histolytica. J Protozool. 1985 Feb;32(1):166–175. doi: 10.1111/j.1550-7408.1985.tb03033.x. [DOI] [PubMed] [Google Scholar]
- McCoy J. J., Mann B. J., Petri W. A., Jr Adherence and cytotoxicity of Entamoeba histolytica or how lectins let parasites stick around. Infect Immun. 1994 Aug;62(8):3045–3050. doi: 10.1128/iai.62.8.3045-3050.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montfort I., Ruiz Argüelles A., Pérez Tamayo R. Phenotypic heterogeneity in the expression of a 30 kDa cysteine proteinase in axenic cultures of Entamoeba histolytica. Arch Med Res. 1992;23(2):99–103. [PubMed] [Google Scholar]
- North M. J., Mottram J. C., Coombs G. H. Cysteine proteinases of parasitic protozoa. Parasitol Today. 1990 Aug;6(8):270–275. doi: 10.1016/0169-4758(90)90189-b. [DOI] [PubMed] [Google Scholar]
- Orozco E., Benitez-Bibriesca L., Hernandez R. Invasion and metastasis mechanisms in Entamoeba histolytica and cancer cells. Some common cellular and molecular features. Mutat Res. 1994 Mar 1;305(2):229–239. doi: 10.1016/0027-5107(94)90243-7. [DOI] [PubMed] [Google Scholar]
- Petri W. A., Jr, Chapman M. D., Snodgrass T., Mann B. J., Broman J., Ravdin J. I. Subunit structure of the galactose and N-acetyl-D-galactosamine-inhibitable adherence lectin of Entamoeba histolytica. J Biol Chem. 1989 Feb 15;264(5):3007–3012. [PubMed] [Google Scholar]
- Petri W. A., Jr, Jackson T. F., Gathiram V., Kress K., Saffer L. D., Snodgrass T. L., Chapman M. D., Keren Z., Mirelman D. Pathogenic and nonpathogenic strains of Entamoeba histolytica can be differentiated by monoclonal antibodies to the galactose-specific adherence lectin. Infect Immun. 1990 Jun;58(6):1802–1806. doi: 10.1128/iai.58.6.1802-1806.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petri W. A., Jr, Mann B. J. Molecular mechanisms of invasion by Entamoeba histolytica. Semin Cell Biol. 1993 Oct;4(5):305–313. doi: 10.1006/scel.1993.1037. [DOI] [PubMed] [Google Scholar]
- Petri W. A., Jr, Smith R. D., Schlesinger P. H., Murphy C. F., Ravdin J. I. Isolation of the galactose-binding lectin that mediates the in vitro adherence of Entamoeba histolytica. J Clin Invest. 1987 Nov;80(5):1238–1244. doi: 10.1172/JCI113198. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petri W. A., Jr, Snodgrass T. L., Jackson T. F., Gathiram V., Simjee A. E., Chadee K., Chapman M. D. Monoclonal antibodies directed against the galactose-binding lectin of Entamoeba histolytica enhance adherence. J Immunol. 1990 Jun 15;144(12):4803–4809. [PubMed] [Google Scholar]
- Ravdin J. I., Croft B. Y., Guerrant R. L. Cytopathogenic mechanisms of Entamoeba histolytica. J Exp Med. 1980 Aug 1;152(2):377–390. doi: 10.1084/jem.152.2.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ravdin J. I. Entamoeba histolytica: from adherence to enteropathy. J Infect Dis. 1989 Mar;159(3):420–429. doi: 10.1093/infdis/159.3.420. [DOI] [PubMed] [Google Scholar]
- Ravdin J. I., Guerrant R. L. Role of adherence in cytopathogenic mechanisms of Entamoeba histolytica. Study with mammalian tissue culture cells and human erythrocytes. J Clin Invest. 1981 Nov;68(5):1305–1313. doi: 10.1172/JCI110377. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ravdin J. I., Jackson T. F., Petri W. A., Jr, Murphy C. F., Ungar B. L., Gathiram V., Skilogiannis J., Simjee A. E. Association of serum antibodies to adherence lectin with invasive amebiasis and asymptomatic infection with pathogenic Entamoeba histolytica. J Infect Dis. 1990 Sep;162(3):768–772. doi: 10.1093/infdis/162.3.768. [DOI] [PubMed] [Google Scholar]
- Ravdin J. I., Murphy C. F. Characterization of the galactose-specific binding activity of a purified soluble Entamoeba histolytica adherence lectin. J Protozool. 1992 Mar-Apr;39(2):319–323. doi: 10.1111/j.1550-7408.1992.tb01322.x. [DOI] [PubMed] [Google Scholar]
- Ravdin J. I., Petri W. A., Murphy C. F., Smith R. D. Production of mouse monoclonal antibodies which inhibit in vitro adherence of Entamoeba histolytica trophozoites. Infect Immun. 1986 Jul;53(1):1–5. doi: 10.1128/iai.53.1.1-5.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reed S., Bouvier J., Pollack A. S., Engel J. C., Brown M., Hirata K., Que X., Eakin A., Hagblom P., Gillin F. Cloning of a virulence factor of Entamoeba histolytica. Pathogenic strains possess a unique cysteine proteinase gene. J Clin Invest. 1993 Apr;91(4):1532–1540. doi: 10.1172/JCI116359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rifkin M. R., Landsberger F. R. Trypanosome variant surface glycoprotein transfer to target membranes: a model for the pathogenesis of trypanosomiasis. Proc Natl Acad Sci U S A. 1990 Jan;87(2):801–805. doi: 10.1073/pnas.87.2.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosales-Encina J. L., Meza I., López-De-León A., Talamás-Rohana P., Rojkind M. Isolation of a 220-kilodalton protein with lectin properties from a virulent strain of Entamoeba histolytica. J Infect Dis. 1987 Nov;156(5):790–797. doi: 10.1093/infdis/156.5.790. [DOI] [PubMed] [Google Scholar]
- Sam-Yellowe T. Y., Shio H., Perkins M. E. Secretion of Plasmodium falciparum rhoptry protein into the plasma membrane of host erythrocytes. J Cell Biol. 1988 May;106(5):1507–1513. doi: 10.1083/jcb.106.5.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sordat B., Piffaretti J. C., Weiss L. Is there a common definition for invasiveness? Report from an informal workshop on Interdisciplinary Aspects of Invasiveness, March 9-11, 1989, Neggio/Lugano, Switzerland. Invasion Metastasis. 1990;10(3):178–192. [PubMed] [Google Scholar]
- Stanley S. L., Jr, Becker A., Kunz-Jenkins C., Foster L., Li E. Cloning and expression of a membrane antigen of Entamoeba histolytica possessing multiple tandem repeats. Proc Natl Acad Sci U S A. 1990 Jul;87(13):4976–4980. doi: 10.1073/pnas.87.13.4976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tannich E., Ebert F., Horstmann R. D. Primary structure of the 170-kDa surface lectin of pathogenic Entamoeba histolytica. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1849–1853. doi: 10.1073/pnas.88.5.1849. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Torian B. E., Lukehart S. A., Stamm W. E. Use of monoclonal antibodies to identify, characterize, and purify a 96,000-dalton surface antigen of pathogenic Entamoeba histolytica. J Infect Dis. 1987 Aug;156(2):334–343. doi: 10.1093/infdis/156.2.334. [DOI] [PubMed] [Google Scholar]
- Tse S. K., Chadee K. Biochemical characterization of rat colonic mucins secreted in response to Entamoeba histolytica. Infect Immun. 1992 Apr;60(4):1603–1612. doi: 10.1128/iai.60.4.1603-1612.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Roy F., Liebaut G., Mareel M., Fiers W. Partial transformation of human tumor cell lines showing defective interaction between unusual p53 gene product and SV40 large-T antigen. Oncogene. 1990 Feb;5(2):207–218. [PubMed] [Google Scholar]
- Vleminckx K. L., Deman J. J., Bruyneel E. A., Vandenbossche G. M., Keirsebilck A. A., Mareel M. M., van Roy F. M. Enlarged cell-associated proteoglycans abolish E-cadherin functionality in invasive tumor cells. Cancer Res. 1994 Feb 15;54(4):873–877. [PubMed] [Google Scholar]
- van Larebeke N. A., Bracke M. E., Mareel M. M. Simple method for quantification of fast plasma membrane movements. Cytometry. 1992;13(1):1–8. doi: 10.1002/cyto.990130103. [DOI] [PubMed] [Google Scholar]