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In vitro Induction of Entamoeba histolytica Cyst-like Structures from Trophozoites

DOI: 10.1371/journal.pntd.0000607

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Abstract:

Inhibition of encystment can be conceived as a potentially useful mechanism to block the transmission of Entamoeba histolytica under natural conditions. Unfortunately, amoeba encystment has not been achieved in vitro and drugs inhibiting the formation of cysts are not available. Luminal conditions inducing encystment in vivo are also unknown, but cellular stress such as exposure to reactive oxygen species from immune cells or intestinal microbiota could be involved. A role for certain divalent cations as cofactors of enzymes involved in excystment has also been described. In this study, we show that trophozoite cultures, treated with hydrogen peroxide in the presence of trace amounts of several cations, transform into small-sized spherical and refringent structures that exhibit resistance to different detergents. Ultrastructural analysis under scanning and transmission electron microscopy revealed multinucleated structures (some with four nuclei) with smooth, thick membranes and multiple vacuoles. Staining with calcofluor white, as well as an ELISA binding assay using wheat germ agglutinin, demonstrated the presence of polymers of N-acetylglucosamine (chitin), which is the primary component of the natural cyst walls. Over-expression of glucosamine 6-phosphate isomerase, likely to be the rate-limiting enzyme in the chitin synthesis pathway, was also confirmed by RT-PCR. These results suggest that E. histolytica trophozoites activated encystment pathways when exposed to our treatment.

References

[1]  WHO (1997) WHO/PAHO/UNESCO report. A consultation with experts on amoebiasis. Mexico City, Mexico, 28–29 January, 1997. WHO Epidemiol Bull. 18. : 13–14.
[2]  Mirelman D, Avron B (1988) Cyst formation in Entamoeba. In: Ravdin JI, editor. Amebiasis: Human infection by Entamoeba histolytica. New York Wiley Medical Publication. pp. 768–781.
[3]  Eichinger D (2001) A role for a galactose lectin and its ligands during encystment of Entamoeba. Eukaryot Microbiol 48 (1): 17–21. doi: 10.1111/j.1550-7408.2001.tb00411.x
[4]  Chávez-Munguía B, Oma?a-Molina M, González-Lázaro M, González-Robles A, Cedillo-Rivera R, et al. (2007) Ultrastructure of cyst differentiation in parasitic protozoa. Parasitol Reserch 100: 1169–1175. doi: 10.1007/s00436-006-0447-x
[5]  López-Romero E, Villagómez-Castro JC (1993) Encystation in Entamoeba invadens. Parasitol Today 9: 225–227. doi: 10.1016/0169-4758(93)90019-C
[6]  Gorb SN (1997) Ultrastructural architecture of the microtrichia of the insect cuticle. J Morphol 234: 1–10. doi: 10.1002/(sici)1097-4687(199710)234:1<1::aid-jmor1>3.0.co;2-i
[7]  Park KE, Kang HK, Lee SJ, Min BM, Park WH (2006) Biomimetic nanofibrous scaffolds: preparation and characterization of PGA/chitin blend nano-fibers. Biomacromolecules 7: 635–643. doi: 10.1021/bm0509265
[8]  Sirijintakarn P, Bailey GB (1980) The relationship of DNA synthesis and cell cycle events to encystation by Entamoeba invadens. Arch Invest Med (Mex) 11: Suppl3–10.
[9]  Clark CG, Espinosa-Cantellano M, Bhattacharya A (2000) Entamoeba histolytica: an overview of the biology of the organism. In: Radvin JI, editor. Amebiasis. Series on Tropical Medicine. London: Imperial College Press. pp. 1–45.
[10]  Ganguly A, Lohia A (2001) The cell cycle of Entamoeba invadens during vegetative growth and differentiation. Mol Biochem Parasitol 112: 277–285. doi: 10.1016/S0166-6851(00)00376-5
[11]  Kane AV, Ward HD, Keusch GT, Pereira ME (1991) In vitro encystation of Giardia lamblia: large-scale production of in vitro cysts and strain and clone differences in encystation efficiency. J Parasitol 77: 974–981. doi: 10.2307/3282752
[12]  Byers TJ, Akins RA, Maynard BJ, Lefken RA, Martin SM (1980) Rapid growth of Acanthamoeba in defined media; induction of encystment by glucose-acetate starvation. J Protozool 27: 216–219. doi: 10.1111/j.1550-7408.1980.tb04684.x
[13]  Vázquezdelara-Cisneros LG, Arroyo-Begovich A (1984) Induction of encystation of Entamoeba invadens by removal of glucose from the culture medium. J Parasitol 70: 629–633. doi: 10.2307/3281741
[14]  Avron B, Stolarsky T, Chayen A, Mirelman D (1986) Encystation of Entamoeba invadens IP-1 is induced by lowering the osmotic pressure and depletion of nutrients from the medium. J Protozool 33: 522–525. doi: 10.1111/j.1550-7408.1986.tb05655.x
[15]  Luján HD, Mowatt MR, Nash TE (1997) Mechanisms of Giardia lamblia differentiation into cysts. Microbiol Mol Biol Rev 61: 294–304.
[16]  Macechko PT, Steimle PA, Lindmark DG, Erlandsen SL, Jaroll EL (1992) Galactosamine-synthesizing enzymes are induced when Giardia encyst. Mol Biochem Parasitol 56: 301–309. doi: 10.1016/0166-6851(92)90179-N
[17]  Morales-Vallarta M, Villarreal-Trevino L, Guerrero-Medrano L, Ramírez-Bon E, Navarro-Marmolejo L, et al. (1997) Entamoeba invadens differentiation and Entamoeba histolytica cyst-like formation induced by CO2. Arch Med Res 28: Suppl150–151.
[18]  González-Salazar F, Viader-Salvadó JM, Martínez-Rodríguez HG, Campos-Góngora E, Mata-Cárdenas BD, et al. (2000) Identification of seven chemical factors that favor high-quality Entamoeba histolytica cyst-like structure formation under axenic conditions. Arch Med Res 31: Suppl192–193. doi: 10.1016/s0188-4409(00)00193-4
[19]  Said-Fernández S, Campos-Góngora E, González-Salazar F, Martínez-Rodríguez HG, Vargas-Villarreal J, et al. (2001) Mg2+, Mn2+, and Co2+ stimulate Entamoeba histolytica to produce chitin-like material. J Parasitol 87: 919–923. doi: 10.1645/0022-3395(2001)087[0919:MMACSE]2.0.CO;2
[20]  Barrón-González MP, Villareal-Trevi?o L, Reséndez-Pérez D, Mata-Cárdenas BD, Morales-Vallarta MR (2008) Entamoeba histolytica: Cyst-like strcutures in vitro induction. Exp Parasitol 118: 600–603. doi: 10.1016/j.exppara.2007.11.002
[21]  Ehrenkaufer GM, Haque R, Hackney JA, Eichinger D, Singh U (2007) Identification of developmentally regulated genes in Entamoeba histolytica: insights into mechanisms of stage conversion in a protozoan parasite. Cel Microbiol 9: 1426–1444. doi: 10.1111/j.1462-5822.2006.00882.x
[22]  Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72: 431–432. doi: 10.1016/0035-9203(78)90144-X
[23]  Ritchie LS (1948) An ether sedimentation technique for routine stool examinations. Bull U S Army Med Dept 8: 326.
[24]  Hamzah Z, Petmitr S, Mungthin M, Leelayoova S, Chavalitshewinkoon-Petmitr P (2006) Differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii by a single-round PCR assay. J Clin Microbiol 44: 3196–3200. doi: 10.1128/JCM.00778-06
[25]  Barrón-González MP, Villareal-Trevi?o L, Verduzco-Martínez JA, Mata-Cárdenas BD, Morales-Vallarta M (2005) Entamoeba invadens: in vitro axenic encystation with a serum substitute. Exp Parasitol 110: 318–321. doi: 10.1016/j.exppara.2005.03.021
[26]  Schupp DG, Januschka MM, Sherlock LA, Stibbs HH, Meyer EA, et al. (1988) Production of viable Giardia cysts in vitro: determination by fluorogenic dye staining, excystation, and animal infectivity in the mouse and Mongolian gerbil. Gastroenterology 95: 1–10.
[27]  Lucero HA, Kuranda MJ, Bulik DA (2002) A nonradioactive, high throughput assay for chitin synthase activity. Anal Biochem 305: 97–105. doi: 10.1006/abio.2002.5594
[28]  Ghosh SK, Field J, Frisardi M, Rosenthal B, Mai Z, et al. (1999) Chitinase secretion by encysting Entamoeba invadens and transfected Entamoeba histolytica trophozoites: localization of secretory vesicles, endoplasmic reticulum, and Golgi apparatus. Infect Immun 67: 3073–3081.
[29]  Eichinger D (2001) Encystation in parasitic protozoa. Curr Opin Microbiol 4: 421–426. doi: 10.1016/S1369-5274(00)00229-0
[30]  Smith AL, Smith HV (1989) A comparison of fluorescein diacetate and propidium iodide staining and in vitro excystation for determining Giardia intestinalis cyst viability. Parasitology 3: 329–331. doi: 10.1017/S0031182000059035
[31]  Campos-Góngora E, Viader-Salvadó JM, Martínez-Rodríguez HG, Zu?iga-Charles MA, Galindo JM, et al. (2000) Mg, Mn, and Co ions enhance the formation of Entamoeba histolytica cyst-like structures resistant to sodium dodecyl sulfate. Arch Med Res 31: 162–8. doi: 10.1016/S0188-4409(00)00054-0
[32]  Nathan CF (1987) Neutrophil activation on biological surfaces. Massive secretion of hydrogen peroxide in response to products of macrophages and lymphocytes. J Clin Invest 80: 1550–1560. doi: 10.1172/JCI113241
[33]  Jourd'heuil D, Morise Z, Conner EM, Grisham MB (1997) Oxidants, transcription factors, and intestinal inflammation. J Clin Gastroenterol 25: Suppl 1S61–72. doi: 10.1097/00004836-199700001-00011
[34]  Halliwell B, Zhao K, Whiteman M (2001) The gastrointestinal tract: a major site of antioxidant action? Free Radic Res 33: 819–830. doi: 10.1080/10715760000301341
[35]  Belozerskaia TA, Gessler NN (2006) Oxidative stress and cell differentiation in Neurospora crassa. Mikrobiologiia 75: 497–501.
[36]  Szymczyk KH, Kerr BA, Freeman TA, Adams CS, Steinbeck MJ (2006) Involvement of hydrogen peroxide in the differentiation and apoptosis of preosteoclastic cells exposed to arsenite. Biochem Pharmacol 72: 761–769. doi: 10.1016/j.bcp.2006.06.027
[37]  Menon J, Rozman R (2007) Oxidative stress, tissue remodeling and regression during amphibian metamorphosis. Comp Biochem Physiol C Toxicol Pharmacol 145: 625–631. doi: 10.1016/j.cbpc.2007.02.011
[38]  Mukherjee C, Clark CG, Lohia A (2008) Entamoeba Shows Reversible Variation in Ploidy under Different Growth Conditions and between Life Cycle Phases. PLoS Negl Trop Dis 2: e281. doi:10.1371/journal.pntd.0000281.
[39]  Arroyo-Begovich A, Cárabez-Trejo A, Ruíz-Herrera J (1980) Identification of the structural component in the cyst wall of Entamoeba invadens. J Parasitol 66: 735–741. doi: 10.2307/3280662
[40]  Arroyo-Begovich A, Cárabez-Trejo A (1982) Location on chitin in the cyst wall of Entamoeba invadens with colloidal gold tracers. J Parasitol 68: 253–258. doi: 10.2307/3281183
[41]  Arroyo-Begovich A, Carabez-Trejo A, de la Torre M (1980) Staining of cysts of Entamoeba invadens, Entamoeba histolytica and Entamoeba coli with wheat germ agglutinin labelled with colloidal gold. Arch Invest Med (Mex) 11: Suppl25–30.
[42]  Van-Keulen H, Steimle PA, Bulik DA, Borowiak RK, Jarroll EL (1998) Cloning of two putative Giardia lamblia glucosamine-6-phosphate isomerase genes only one which is transcriptionally activated during encystment. J Eukaryot Microbiol 45: 637–642. doi: 10.1111/j.1550-7408.1998.tb04560.x
[43]  López AB, Hossain MT, Van-Keulen H (2002) Giardia intestinalis glucosamine 6-phosphate isomerase: the key enzyme to encystment appers to be controlled by ubiquitin attachment. J Eukaryot Microbiol 49: 134–136. doi: 10.1111/j.1550-7408.2002.tb00356.x
[44]  Loftus B, Anderson I, Davies R, Alsmark UCM, Samuelson J, et al. (2005) The genome of the protist parasite Entamoeba histolytica. Nature 433: 865–868. doi: 10.1038/nature03291
[45]  Jarroll EL, Macechko PT, Steimle PA, Bulik D, Karr CD, et al. (2001) Regulation of carbohydrate metabolism during Giardia encystment. J Eukaryot Microbiol 48: 22–26. doi: 10.1111/j.1550-7408.2001.tb00412.x
[46]  Vicente JB, Ehrenkaufer GM, Saraiva LM, Teixeira M, Singh U (2009) Entamoeba histolytica modulates a complex repertoire of novel genes in response to oxidative and nitrosative stresses: implications for amebic pathogenesis. Cell Microbiol 11: 51–69. doi: 10.1111/j.1462-5822.2008.01236.x
[47]  Campos-Góngora E, Viader-Salvadó JM, Martínez-Rodríguez HG, Mora-Galindo J, Said-Fernández S (1997) Stimulation of Entamoeba histolytica cyst wall polysaccharide synthesis by three divalent cations. Arch Med Res 28 (Spec No): 141–142.
[48]  Campos-Góngora E, Ebert F, Willhoeft U, Said-Fernández S, Tannich E (2004) Charaterization of chitin synthases from Entamoeba. Protist 155: 323–330. doi: 10.1078/1434461041844204
[49]  Karr CD, Jarroll EL (2004) Cyst wall synthase: N-acetylgalactosaminyltransferase activity is induced to form the novel N-acetylgalactosamine polysaccharide in the Giardia cyst wall. Microbiology 150: 1237–1243. doi: 10.1099/mic.0.26922-0
[50]  Takano J, Narita T, Tachibana H, Shimizu T, Komatsubara H, et al. (2005) Entamoeba histolytica and Entamoeba dispar infections in cynomolgus monkeys imported into Japan for research. Parasitol Res 97: 255–257. doi: 10.1007/s00436-005-1415-6
[51]  Leitch GJ, Udezulu IA, He Q, Visvesvara GS (1993) Effects of protein malnutrition on experimental Giardiasis in the Mongolian gerbil. Scand J Gastroenterol 28: 885–893. doi: 10.3109/00365529309103130
[52]  Yanke SJ, Ceri H, McAllister TA, Morck DW, Olson ME (1998) Serum immune response to Giardia duodenalis in experimentally infected lambs. Vet Parasitol 75: 9–19. doi: 10.1016/S0304-4017(97)00187-8
[53]  Larocque R, Nakagaki K, Lee P, Abdul-Wahid A, Faubert GM (2003) Immunization of BALB/c mice with Giardia duodenalis recombinant cyst wall protein inhibits shedding of cysts. Infect Immun 71: 5662–5669. doi: 10.1128/IAI.71.10.5662-5669.2003
[54]  Stein JE, Radecki SV, Lappin MR (2003) Efficacy of Giardia vaccination in the treatment of giardiasis in cats. J Am Vet Med Assoc 222: 1548–1551. doi: 10.2460/javma.2003.222.1548

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