Differential Expression of Pathogenic Genes of Entamoeba Histolytica Vs E. Dispar in a Model of Infection Using Human Liver Tissue Explants

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Aug 4

PMID 28771523

Citations 12

Authors

Cecilia Ximenez

Enrique Gonzalez

Miriam Nieves

Ulises Magana

Patricia Moran

Marco Gudino-Zayas

Oswaldo Partida

Eric Hernandez

Liliana Rojas-Velazquez

Ma Carmen Garcia de Leon

Hector Maldonado

Affiliations

Soon will be listed here.

Abstract

We sought to establish an ex vivo model for examining the interaction of E. histolytica with human tissue, using precision-cut liver slices (PCLS) from donated organs. E. histolytica- or E. dispar-infected PCLS were analyzed at different post-infection times (0, 1, 3, 24 and 48 h) to evaluate the relation between tissue damage and the expression of genes associated with three factors: a) parasite survival (peroxiredoxin, superoxide dismutase and 70 kDa heat shock protein), b) parasite virulence (EhGal/GalNAc lectin, amoebapore, cysteine proteases and calreticulin), and c) the host inflammatory response (various cytokines). Unlike E. dispar (non-pathogenic), E. histolytica produced some damage to the structure of hepatic parenchyma. Overall, greater expression of virulence genes existed in E. histolytica-infected versus E. dispar-infected tissue. Accordingly, there was an increased expression of EhGal/GalNAc lectin, Ehap-a and Ehcp-5, Ehcp-2, ehcp-1 genes with E. histolytica, and a decreased or lack of expression of Ehcp-2, and Ehap-a genes with E. dispar. E. histolytica-infected tissue also exhibited an elevated expression of genes linked to survival, principally peroxiredoxin, superoxide dismutase and Ehhsp-70. Moreover, E. histolytica-infected tissue showed an overexpression of some genes encoding for pro-inflammatory interleukins (ILs), such as il-8, ifn-γ and tnf-α. Contrarily, E. dispar-infected tissue displayed higher levels of il-10, the gene for the corresponding anti-inflammatory cytokine. Additionally, other genes were investigated that are important in the host-parasite relationship, including those encoding for the 20 kDa heat shock protein (HSP-20), the AIG-1 protein, and immune dominant variable surface antigen, as well as for proteins apparently involved in mechanisms for the protection of the trophozoites in different environments (e.g., thioredoxin-reductase, oxido-reductase, and 9 hypothetical proteins). Some of the hypothetical proteins evidenced interesting overexpression rates, however we should wait to their characterization. This finding suggest that the present model could be advantageous for exploring the complex interaction between trophozoites and hepatocytes during the development of ALA, particularly in the initial stages of infection.

Citing Articles

Attenuation of In Vitro and In Vivo Virulence Is Associated with Repression of Gene Expression of AIG1 Gene in .

Lozano-Mendoza J, Ramirez-Montiel F, Rangel-Serrano A, Paramo-Perez I, Mendoza-Macias C, Saavedra-Salazar F Pathogens. 2023; 12(3).

PMID: 36986411 PMC: 10051847. DOI: 10.3390/pathogens12030489.

Proteases as Therapeutic Targets Against the Parasitic Cnidarian : Characterization of Molecules Key to Parasite Virulence In Salmonid Hosts.

Alama-Bermejo G, Bartosova-Sojkova P, Atkinson S, Holzer A, Bartholomew J Front Cell Infect Microbiol. 2022; 11:804864.

PMID: 35071050 PMC: 8777295. DOI: 10.3389/fcimb.2021.804864.

The Dichloromethane Fraction of , A Plant Used in Sonoran Traditional Medicine, Affect Erythrophagocytosis and Gene Expression.

Villegas-Gomez I, Silva-Olivares A, Robles-Zepeda R, Galvez-Ruiz J, Shibayama M, Valenzuela O Front Cell Infect Microbiol. 2021; 11:693449.

PMID: 34368014 PMC: 8343225. DOI: 10.3389/fcimb.2021.693449.

: Proteomics Bioinformatics Reveal Predictive Functions and Protein-Protein Interactions of Differentially Abundant Membrane and Cytosolic Proteins.

Azmi N, Othman N Membranes (Basel). 2021; 11(6).

PMID: 34063994 PMC: 8224062. DOI: 10.3390/membranes11060376.

The Interactions of Parasite Calreticulin With Initial Complement Components: Consequences in Immunity and Virulence.

Ramirez-Toloza G, Aguilar-Guzman L, Valck C, Ferreira V, Ferreira A Front Immunol. 2020; 11:1561.

PMID: 32793217 PMC: 7391170. DOI: 10.3389/fimmu.2020.01561.

References

Bruchhaus I, Jacobs T, Leippe M, Tannich E . Entamoeba histolytica and Entamoeba dispar: differences in numbers and expression of cysteine proteinase genes. Mol Microbiol. 1996; 22(2):255-63. DOI: 10.1046/j.1365-2958.1996.00111.x. View

Ocadiz-Delgado R, Marroquin-Chavira A, Hernandez-Mote R, Valencia C, Manjarrez-Zavala M, Covarrubias L . Induction of focal epithelial hyperplasia in tongue of young bk6-E6/E7 HPV16 transgenic mice. Transgenic Res. 2009; 18(4):513-27. DOI: 10.1007/s11248-009-9243-6. View

Leippe M . Amoebapores. Parasitol Today. 1997; 13(5):178-83. DOI: 10.1016/s0169-4758(97)01038-7. View

Braga L, Ninomiya H, McCoy J, Eacker S, Wiedmer T, Pham C . Inhibition of the complement membrane attack complex by the galactose-specific adhesion of Entamoeba histolytica. J Clin Invest. 1992; 90(3):1131-7. PMC: 329975. DOI: 10.1172/JCI115931. View

Lopez-Vancell R, Montfort I, Perez-Tamayo R . Galactose-specific adhesin and cytotoxicity of Entamoeba histolytica. Parasitol Res. 2000; 86(3):226-31. DOI: 10.1007/s004360050035. View

Andra J, Herbst R, Leippe M . Amoebapores, archaic effector peptides of protozoan origin, are discharged into phagosomes and kill bacteria by permeabilizing their membranes. Dev Comp Immunol. 2003; 27(4):291-304. DOI: 10.1016/s0145-305x(02)00106-4. View

Olivos-Garcia A, Nequiz-Avendano M, Tello E, Martinez R, Gonzalez-Canto A, Lopez-Vancell R . Inflammation, complement, ischemia and amoebic survival in acute experimental amoebic liver abscesses in hamsters. Exp Mol Pathol. 2004; 77(1):66-71. DOI: 10.1016/j.yexmp.2003.09.003. View

Meyer M, Fehling H, Matthiesen J, Lorenzen S, Schuldt K, Bernin H . Overexpression of Differentially Expressed Genes Identified in Non-pathogenic and Pathogenic Entamoeba histolytica Clones Allow Identification of New Pathogenicity Factors Involved in Amoebic Liver Abscess Formation. PLoS Pathog. 2016; 12(8):e1005853. PMC: 5004846. DOI: 10.1371/journal.ppat.1005853. View

Loftus B, Anderson I, Davies R, Alsmark U, Samuelson J, Amedeo P . The genome of the protist parasite Entamoeba histolytica. Nature. 2005; 433(7028):865-8. DOI: 10.1038/nature03291. View

10.

Nuovo G . The utility of in situ--based methodologies including in situ polymerase chain reaction for the diagnosis and study of viral infections. Hum Pathol. 2007; 38(8):1123-36. DOI: 10.1016/j.humpath.2007.04.005. View

11.

Michalak M, Corbett E, Mesaeli N, Nakamura K, Opas M . Calreticulin: one protein, one gene, many functions. Biochem J. 1999; 344 Pt 2:281-92. PMC: 1220642. View

12.

MacFarlane R, Singh U . Identification of differentially expressed genes in virulent and nonvirulent Entamoeba species: potential implications for amebic pathogenesis. Infect Immun. 2005; 74(1):340-51. PMC: 1346599. DOI: 10.1128/IAI.74.1.340-351.2006. View

13.

Yalcin A . Quantification of thioredoxin mRNA expression in the rat hippocampus by real-time PCR following oxidative stress. Acta Biochim Pol. 2004; 51(4):1059-65. View

14.

Choi M, Sajed D, Poole L, Hirata K, Herdman S, Torian B . An unusual surface peroxiredoxin protects invasive Entamoeba histolytica from oxidant attack. Mol Biochem Parasitol. 2005; 143(1):80-9. DOI: 10.1016/j.molbiopara.2005.04.014. View

15.

Ajuebor M, Swain M, Perretti M . Chemokines as novel therapeutic targets in inflammatory diseases. Biochem Pharmacol. 2002; 63(7):1191-6. DOI: 10.1016/s0006-2952(02)00854-7. View

16.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

17.

Ximenez C, Gonzalez E, Nieves M, Silva-Olivares A, Shibayama M, Galindo-Gomez S . Entamoeba histolytica and E. dispar Calreticulin: inhibition of classical complement pathway and differences in the level of expression in amoebic liver abscess. Biomed Res Int. 2014; 2014:127453. PMC: 4016843. DOI: 10.1155/2014/127453. View

18.

Carranza-Rosales P, Santiago-Mauricio M, Guzman-Delgado N, Vargas-Villarreal J, Lozano-Garza G, Viveros-Valdez E . Induction of virulence factors, apoptosis, and cytokines in precision-cut hamster liver slices infected with Entamoeba histolytica. Exp Parasitol. 2012; 132(4):424-33. DOI: 10.1016/j.exppara.2012.09.012. View

19.

Rastew E, Vicente J, Singh U . Oxidative stress resistance genes contribute to the pathogenic potential of the anaerobic protozoan parasite, Entamoeba histolytica. Int J Parasitol. 2012; 42(11):1007-15. PMC: 3483436. DOI: 10.1016/j.ijpara.2012.08.006. View

20.

Graaf I, Groothuis G, Olinga P . Precision-cut tissue slices as a tool to predict metabolism of novel drugs. Expert Opin Drug Metab Toxicol. 2007; 3(6):879-98. DOI: 10.1517/17425255.3.6.879. View