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. 1990 Sep 1;172(3):879–888. doi: 10.1084/jem.172.3.879

Characterization of an immuno-dominant variable surface antigen from pathogenic and nonpathogenic Entamoeba histolytica

PMCID: PMC2188528  PMID: 1696956

Abstract

A 125-kD surface antigen of Entamoeba histolytica is recognized by 73% of immune sera from patients with amoebic liver abscesses. Using pooled human immune sera a cDNA clone (lambda cM17) encoding this antigen (M17) has been isolated from a lambda gt11 expression library of the virulent stain E. histolytica HM1:IMSS. Monospecific antibodies, purified by binding to phage lysate of lambda cM17, and mAb FA7 reacted exclusively with the 125-kD antigen by Western blot analysis. Surface binding and cap formation are observed with patient sera, purified monospecific antiserum, and mAb FA7. Corresponding genomic clones (pBSgM17-1/2/3) were isolated by hybridization with the cDNA clone. These contained an open-reading frame of 3345 bp, which is in good agreement with the mRNA size of approximately 3.0 kb as revealed by Northern hybridization with lambda cM17. The inferred amino acid sequence predicts a 125,513 dalton protein that contains 17 potential N- linked glycosylation sites and is unusually rich in tyrosine and asparagine residues. A distinctly hydrophobic NH2-terminal region may serve as membrane anchor or signal sequence. In contrast to conservation of an immunodominant epitope recognized in pathogenic and nonpathogenic strains by monoclonal FA7 and human immune sera, amplification and sequence analysis of a 1,4000-bp fragment of this gene from a fresh nonpathogenic isolate by use of the PCR demonstrate regions of significant sequence divergence in this antigen. A 1% sequence variability among different isolates of the pathogenic strain HM1:IMSS and a 12-13% variability between pathogenic and nonpathogenic strains are revealed by comparison to published partial amino acid sequences (Tannich, E., R.D. Horstmann, J. Knobloch, and H.H. Arnold. 1989. Proc. Natl. Acad. Sci. USA. 86:5118). Some restriction enzymes were found that allowed PCR diagnosis of nonpathogenic and pathogenic isolates with the exclusion of E. histolytica-like Laredo, suggesting that a detailed study of nonpathogenic and pathogenic isolates in relation to the M17 antigen sequence will provide a basis of differentiating isolates.

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Selected References

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  1. Aley S. B., Scott W. A., Cohn Z. A. Plasma membrane of Entamoeba histolytica. J Exp Med. 1980 Aug 1;152(2):391–404. doi: 10.1084/jem.152.2.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chadee K., Johnson M. L., Orozco E., Petri W. A., Jr, Ravdin J. I. Binding and internalization of rat colonic mucins by the galactose/N-acetyl-D-galactosamine adherence lectin of Entamoeba histolytica. J Infect Dis. 1988 Aug;158(2):398–406. doi: 10.1093/infdis/158.2.398. [DOI] [PubMed] [Google Scholar]
  3. Chadee K., Petri W. A., Jr, Innes D. J., Ravdin J. I. Rat and human colonic mucins bind to and inhibit adherence lectin of Entamoeba histolytica. J Clin Invest. 1987 Nov;80(5):1245–1254. doi: 10.1172/JCI113199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Edman U., Meza I., Agabian N. Genomic and cDNA actin sequences from a virulent strain of Entamoeba histolytica. Proc Natl Acad Sci U S A. 1987 May;84(9):3024–3028. doi: 10.1073/pnas.84.9.3024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garfinkel L. I., Giladi M., Huber M., Gitler C., Mirelman D., Revel M., Rozenblatt S. DNA probes specific for Entamoeba histolytica possessing pathogenic and nonpathogenic zymodemes. Infect Immun. 1989 Mar;57(3):926–931. doi: 10.1128/iai.57.3.926-931.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gillin F. D., Diamond L. S. Clonal growth of Entamoeba histolytica and other species of Entamoeba in agar. J Protozool. 1978 Nov;25(4):539–543. doi: 10.1111/j.1550-7408.1978.tb04182.x. [DOI] [PubMed] [Google Scholar]
  8. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  9. Huber M., Garfinkel L., Gitler C., Mirelman D., Revel M., Rozenblatt S. Entamoeba histolytica: cloning and characterization of actin cDNA. Mol Biochem Parasitol. 1987 Jul;24(3):227–235. doi: 10.1016/0166-6851(87)90154-x. [DOI] [PubMed] [Google Scholar]
  10. Huber M., Garfinkel L., Gitler C., Mirelman D., Revel M., Rozenblatt S. Nucleotide sequence analysis of an Entamoeba histolytica ferredoxin gene. Mol Biochem Parasitol. 1988 Oct;31(1):27–33. doi: 10.1016/0166-6851(88)90142-9. [DOI] [PubMed] [Google Scholar]
  11. Keene W. E., Petitt M. G., Allen S., McKerrow J. H. The major neutral proteinase of Entamoeba histolytica. J Exp Med. 1986 Mar 1;163(3):536–549. doi: 10.1084/jem.163.3.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meza I., Cázares F., Rosales-Encina J. L., Talamás-Rohana P., Rojkind M. Use of antibodies to characterize a 220-kilodalton surface protein from Entamoeba histolytica. J Infect Dis. 1987 Nov;156(5):798–805. doi: 10.1093/infdis/156.5.798. [DOI] [PubMed] [Google Scholar]
  13. Meza I., De la Garza I. M., Meraz M. A., Gallegos B., De la Torre M., Tanimoto M., Martínez-Palomo A. Isoenzyme patterns of Entamoeba histolytica isolates from asymptomatic carriers: use of gradient acrylamide gels. Am J Trop Med Hyg. 1986 Nov;35(6):1134–1139. doi: 10.4269/ajtmh.1986.35.1134. [DOI] [PubMed] [Google Scholar]
  14. Meza I., Sabanero M., Cazares F., Bryan J. Isolation and characterization of actin from Entamoeba histolytica. J Biol Chem. 1983 Mar 25;258(6):3936–3941. [PubMed] [Google Scholar]
  15. Mirelman D. Ameba-bacterium relationship in amebiasis. Microbiol Rev. 1987 Jun;51(2):272–284. doi: 10.1128/mr.51.2.272-284.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mirelman D., Bracha R., Chayen A., Aust-Kettis A., Diamond L. S. Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence. Exp Parasitol. 1986 Aug;62(1):142–148. doi: 10.1016/0014-4894(86)90017-2. [DOI] [PubMed] [Google Scholar]
  17. Mirelman D. Effect of culture conditions and bacterial associates on the zymodemes of Entamoeba histolytica. Parasitol Today. 1987 Feb;3(2):37–37. doi: 10.1016/0169-4758(87)90210-9. [DOI] [PubMed] [Google Scholar]
  18. Morgan D. O., Edman J. C., Standring D. N., Fried V. A., Smith M. C., Roth R. A., Rutter W. J. Insulin-like growth factor II receptor as a multifunctional binding protein. Nature. 1987 Sep 24;329(6137):301–307. doi: 10.1038/329301a0. [DOI] [PubMed] [Google Scholar]
  19. Moss D. M., Mathews H. M. A fast electrophoretic isoenzyme technique for the identification of invasive and non-invasive Entamoeba histolytica and "E. histolytica-like" organisms. J Protozool. 1987 Aug;34(3):253–255. doi: 10.1111/j.1550-7408.1987.tb03169.x. [DOI] [PubMed] [Google Scholar]
  20. Petri W. A., Jr, Broman J., Healy G., Quinn T., Ravdin J. I. Antigenic stability and immunodominance of the Gal/GalNAc adherence lectin of Entamoeba histolytica. Am J Med Sci. 1989 Mar;297(3):163–165. doi: 10.1097/00000441-198903000-00006. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Reed S. L., Keene W. E., McKerrow J. H., Gigli I. Cleavage of C3 by a neutral cysteine proteinase of Entamoeba histolytica. J Immunol. 1989 Jul 1;143(1):189–195. [PubMed] [Google Scholar]
  23. Samuelson J., Acuna-Soto R., Reed S., Biagi F., Wirth D. DNA hybridization probe for clinical diagnosis of Entamoeba histolytica. J Clin Microbiol. 1989 Apr;27(4):671–676. doi: 10.1128/jcm.27.4.671-676.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sargeaunt P. G. The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasitol Today. 1987 Feb;3(2):40–37. doi: 10.1016/0169-4758(87)90211-0. [DOI] [PubMed] [Google Scholar]
  25. Sargeaunt P. G., Williams J. E. Electrophoretic isoenzyme patterns of the pathogenic and non-pathogenic intestinal amoebae of man. Trans R Soc Trop Med Hyg. 1979;73(2):225–227. doi: 10.1016/0035-9203(79)90219-0. [DOI] [PubMed] [Google Scholar]
  26. Sargeaunt P. G., Williams J. E., Grene J. D. The differentiation of invasive and non-invasive Entamoeba histolytica by isoenzyme electrophoresis. Trans R Soc Trop Med Hyg. 1978;72(5):519–521. doi: 10.1016/0035-9203(78)90174-8. [DOI] [PubMed] [Google Scholar]
  27. Sargeaunt P. G., Williams J. E., Neal R. A. A comparative study of Entamoeba histolytica (NIH :200, HK9, etc.), "E. histolytica-like" and other morphologically identical amoebae using isoenzyme electrophoresis. Trans R Soc Trop Med Hyg. 1980;74(4):469–474. doi: 10.1016/0035-9203(80)90058-9. [DOI] [PubMed] [Google Scholar]
  28. Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Strachan W. D., Chiodini P. L., Spice W. M., Moody A. H., Ackers J. P. Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica. Lancet. 1988 Mar 12;1(8585):561–563. doi: 10.1016/s0140-6736(88)91355-4. [DOI] [PubMed] [Google Scholar]
  30. Tannich E., Horstmann R. D., Knobloch J., Arnold H. H. Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5118–5122. doi: 10.1073/pnas.86.13.5118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Torian B. E., Reed S. L., Flores B. M., Creely C. M., Coward J. E., Vial K., Stamm W. E. The 96-kilodalton antigen as an integral membrane protein in pathogenic Entamoeba histolytica: potential differences in pathogenic and nonpathogenic isolates. Infect Immun. 1990 Mar;58(3):753–760. doi: 10.1128/iai.58.3.753-760.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Torian B. E., Reed S. L., Flores B. M., Plorde J., Stamm W. E. Serologic response to the 96,000-Da surface antigen of pathogenic Entamoeba histolytica. J Infect Dis. 1989 Apr;159(4):794–797. doi: 10.1093/infdis/159.4.794. [DOI] [PubMed] [Google Scholar]
  34. Weinberger C., Hollenberg S. M., Ong E. S., Harmon J. M., Brower S. T., Cidlowski J., Thompson E. B., Rosenfeld M. G., Evans R. M. Identification of human glucocorticoid receptor complementary DNA clones by epitope selection. Science. 1985 May 10;228(4700):740–742. doi: 10.1126/science.2581314. [DOI] [PubMed] [Google Scholar]
  35. Young J. D., Young T. M., Lu L. P., Unkeless J. C., Cohn Z. A. Characterization of a membrane pore-forming protein from Entamoeba histolytica. J Exp Med. 1982 Dec 1;156(6):1677–1690. doi: 10.1084/jem.156.6.1677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]

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