The Monocyte Locomotion Inhibitory Factor (MLIF) Produced by Entamoeba histolytica Alters the Expression of Genes Related to the Wound-Healing Process

  • R. Silva-García
  • M. E. Morales-Martínez
  • F. Blanco-Favela
  • A. Torres-Salazar
  • J. Ríos-Olvera
  • M. R. Garrido-Ortega
  • E. Tesoro-Cruz
  • G. Rico-Rosillo


The monocyte locomotion inhibitory factor (MLIF) is a pentapeptide produced by Entamoeba histolytica. This factor displays several in vivo and in vitro anti-inflammatory properties, among others, inhibition of monocyte locomotion and the respiratory burst in monocyte and neutrophils. A synthetic peptide had the same selective anti-inflammatory features as the native material. We now evaluated MLIF on the constitutive and induced gene expression in the MRC-5 human fibroblasts cell line. The MLIF affected constitutive expression of 21 genes and induced expression of 75 genes. Some of these genes involved in the inflammatory response and other participated in the remodeling and the wound-healing processes.


Monocyte locomotion inhibitory factor Entamoeba histolytica Inflammation Anti-inflammatory pentapeptide Microarrays Gene expression Cytokines 



The authors wished to thank Diego Adrián Oliva Rico from the Licenciatura en Investigación Biomédica, UNAM, for the comments and critical review of the manuscript.


  1. Al-Shahrour F, Minguez F, Vaqueriza J, Conde L, Dopazo J (2005) BABELOMICS: a suite of web-tools for functional annotation and analysis of group of genes in high-throughput experiments. Nucleic Acids Res 33:W460–W464PubMedCrossRefGoogle Scholar
  2. Balemans W, Van HW (2002) Extracellular regulation of BMP signaling in vertebrates: a cocktail of modulators. Dev Biol 250:231–250PubMedCrossRefGoogle Scholar
  3. Bilezikjian LM, Blount AL, Donaldson CJ, Vale WW (2006) Pituitary actions of ligands of the TGF-beta family: activins and inhibins. Reproduction 132:207–215PubMedCrossRefGoogle Scholar
  4. Busold C, Winter S, Hauser N, Bauer A et al (2005) Integration of GO annotations in correspondence analysis: facilitating the interpretation of microarray data. Bioinformatics 21:2424–2429PubMedCrossRefGoogle Scholar
  5. Cárdenas MG, Garay A, Rico G, Giménez-Scherer JA, Kretschmer R (2002) Reduction of peritoneal adherences by the anti-inflammatory peptide produced by E. histolytica in gerbil model. Clin Immunol 103:34–35CrossRefGoogle Scholar
  6. Carmeliet P, Ferreira V, Breier G, Pollefeyt S et al (1996) Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380:435–439PubMedCrossRefGoogle Scholar
  7. Chae JK, Kim I, Lim ST, Chung MJ et al (2000) Coadministration of angiopoietin-1 and vascular endothelial growth factor enhances collateral vascularization. Arterioscler Thromb Vasc Biol 20:2573–2578PubMedCrossRefGoogle Scholar
  8. Clauss M (2000) Molecular biology of the VEGF and VEGF receptor family. Semin Thromb Hemost 26:561–569PubMedCrossRefGoogle Scholar
  9. deBleser PJ, Niki T, Xu G, Rogiers V, Geerts A (1997) Localization and cellular sources of activins in normal and fibrotic rat liver. Hepatology 26:905–912CrossRefGoogle Scholar
  10. deKretser DM, Hedger MP, Phillips DJ (1999) Activin A and follistatin: their role in the acute phase reaction and inflammation. J Endocrinol 161:195–198CrossRefGoogle Scholar
  11. Díaz UR, al-Shahrour F, Dopazo J (2004) The use of GO terms to understand the biological significance of microarray differential gene expression DATA. In: Kimberly F, Simon M (eds). Kluwer Academic Publishers, New York, pp 233–247Google Scholar
  12. Drexler HG (1996) Expression of FLT3 receptor and response to FLT3 ligand by leukemic cells. Leukemia 10:588–599PubMedGoogle Scholar
  13. Dunk C, Shams M, Nijjar S, Rhaman M et al (2000) Angiopoietin-1 and angiopoietin-2 active trophoblast Tie-2 to promote growth and migration during placental development. Am J Pathol 156:2185–2199PubMedCrossRefGoogle Scholar
  14. Emanueli C, Schratzberger P, Kirchmair R, Madeddu P (2003) Paracrine control of vascularization and neurogenesis by neurotrophins. Br J Pharmacol 140:614–619PubMedCrossRefGoogle Scholar
  15. Gale NW, Yancopoulos GD (1999) Growth factors acting via endothelial cell-specific receptor tyrosine kinases: VEGFs, angiopoietins, and ephrins in vascular development. Genes Dev 13:1055–1066PubMedCrossRefGoogle Scholar
  16. Giménez-Scherer JA, Pacheco Cano MG, Cruz de Lavin E, Hernández-Jauregui P, Merchant MT, Kretschmer RR (1987) Ultrastructural changes associated with the inhibition of monocyte chemotaxis caused by products of axenically grown Entamoeba histolytica. Lab Investig 57:45–51PubMedGoogle Scholar
  17. Giménez-Scherer JA, Rico G, Fernández-Díez J, Kretschmer R (1997) Inhibition of contact cutaneous delayed hypersensitivity reactions to DNCB in guinea pigs by the Monocyte Locomotion Inhibitory Factor (MLIF) produced by axenically grown Entamoeba histolytica. Arch Med Res 28:237–238PubMedGoogle Scholar
  18. Giménez-Scherer JA, Arenas E, Díaz L, Rico G, Fernández J et al (2000) Effect of the monocyte locomotion inhibitory factor (MLIF) produced by Entamoeba histolytica on the expression of cell adhesion molecules (CAMs) in the skin of Guinea pigs. Arch Med Res 31:92–93CrossRefGoogle Scholar
  19. Gumbiner BM (1996) Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell 84:345–357PubMedCrossRefGoogle Scholar
  20. Hanahan D (1997) Signaling vascular morphogenesis and maintenance. Science 277:48–50PubMedCrossRefGoogle Scholar
  21. Hernández-Rodríguez J, Segarra M, Vilardell C, Sánchez M et al (2003) Elevated production of interleukin-6 is associated with a lower incidence of disease-related ischemic events in patients with giant-cell arteritis: angiogenic activity of interleukin-6 as a potential protective mechanism. Circulation 107:2428–2434PubMedCrossRefGoogle Scholar
  22. Himanen JP, Nikolov DB (2003) Eph receptors and ephrins. Int J Biochem Cell Biol 35:130–134PubMedCrossRefGoogle Scholar
  23. Hsu HC, Tsai WH, Jiang ML, Ho CH et al (1999) Circulating levels of thrombopoietic and inflammatory cytokines in patients with clonal and reactive thrombocytosis. J Lab Clin Med 134:392–397PubMedCrossRefGoogle Scholar
  24. Hubner G, Hu Q, Smola H, Werner S (1996) Strong induction of activin expression after injury suggests an important role of activin in wound repair. Dev Biol 173:490–498PubMedCrossRefGoogle Scholar
  25. Hubner G, Brauchle M, Gregor M, Werner S (1997) Activin A: a novel player and inflammatory marker in inflammatory bowel disease. Lab Investig 77:311–318PubMedGoogle Scholar
  26. Huynh-Do U, Stein E, Lane AA, Liu H et al (1999) Surface densities of ephrin-B1 determine EphB1-coupled activation of cell attachment through alpha5beta3 and alpha5beta1 integrins. EMBO J 18:2165–2173PubMedCrossRefGoogle Scholar
  27. Ivanov AI, Romanovsky A (2006) Putative dual role of ephrin–Eph receptor interactions in inflammation. IUBMB Life 58:389–394PubMedCrossRefGoogle Scholar
  28. Jordana M, Sarnstrand B, Sime P, Ramis I (1994) Immune-inflammatory functions of fibroblasts. Eur Respir J 7:2212–2222PubMedCrossRefGoogle Scholar
  29. Kozian DH, Ziche M, Hellmut GA (1997) The activin-binding protein follistatin regulates autocrine endothelial cell activity and induces angiogenesis. Lab Investig 76:267–276PubMedGoogle Scholar
  30. Kretschmer R, Collado M, Pacheco G, Salinas M et al (1985) Inhibition of human monocyte locomotion by products of axenically grown Entamoeba histolytica. Parasite Immunol 5:527–543CrossRefGoogle Scholar
  31. Kretschmer R, Rico G, Giménez-Scherer JA (2001) A novel anti-inflammatory oligopeptide produced by Entamoeba histolyitica. Mol Biochem Parasitol 112:201–209PubMedCrossRefGoogle Scholar
  32. Lamont AG, Adorini L (1996) IL-12: a key cytokine in immune regulation. Immunol Today 17:214–217PubMedCrossRefGoogle Scholar
  33. Lin CY, Yang YH, Lee CC, Huang CL et al (2006) Thrombopoietin and interleukin-6 levels in Henoch-Schonlein purpura. J Microbiol Immunol Infect 39:476–482PubMedGoogle Scholar
  34. Maisonpierre PC, Suri C, Jones PF, Bartunkova S et al (1997) Angiopoietin-2 a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science 277:55–60PubMedCrossRefGoogle Scholar
  35. Mamluk R, Klagsbrun M, Detmar M, Bielenberg D (2005) Soluble neurophilin targeted to the skin inhibits vascular permeability. Angiogenesis 8:217–227PubMedCrossRefGoogle Scholar
  36. Mashasiro S, Masayuki K, Takefumi I, Akiko W et al (2000) Differential regulation of metalloproteinase production, proliferation and chemotaxis of human lung fibroblasts by PDGF, interleukin-1β and TNF-α. Mediat Inflamm 9:1551–1560Google Scholar
  37. Mehrnaz G, Sem HP (2001) Role of cytokines and cytokine therapy in wound healing and fibrotic disease. Curr Pharm Des 7:1083–1103CrossRefGoogle Scholar
  38. Munz B, Hubner G, Tretter Y, Alzheimer C et al (1999) A novel role of activin in inflammation and repair. J Endocrinol 161:187–193PubMedCrossRefGoogle Scholar
  39. Murai KK, Pasquale EB (2003) “Eph”ective signaling: forward reverse and crosstalk. J Cell Sci 116:2823–2832PubMedCrossRefGoogle Scholar
  40. Muraille E, Leo O (1998) Revisiting the Th1/Th2 paradigm. Scand J Immunol 47:1–9PubMedCrossRefGoogle Scholar
  41. Ohguchi M, Yamato K, Ishihara Y, Koide M et al (1998) Activin A regulates the production of mature interleukin-1beta and interleukin-1 receptor antagonist in human monocytic cells. J Interferon Cytokine Res 18:491–498PubMedCrossRefGoogle Scholar
  42. Ohmizono Y, Sakabe H, Kimura T, Tanimukai S et al (1997) Thrombopoietin augments ex vivo expansion of human cord blood-derived hematopoietic progenitors in combination with stem cell factor and flt3 ligand. Leukemia 11:524–530PubMedCrossRefGoogle Scholar
  43. Papapetropoulos A, Garcia-Cardena G, Dengler TJ, Maisonpierre PC et al (1999) Direct actions of angiopoietin-1 on human endothelium: evidence for network stabilization, cell survival, and interaction with other angiogenic growth factors. Lab Investig 79:213–223PubMedGoogle Scholar
  44. Pérez-Tamayo R, Brandt H (1971) Amebiasis. In: Marcial R (ed) Pathology of protozoan and helminthic disease. Williams and Wilkins, Baltimore, pp 145–188Google Scholar
  45. Phillip DJ, Jones KL, Scheerlinck JY, Hedger MP et al (2001) Evidence for activin A and follistatin involvement in the systemic inflammatory response. Mol Cell Endocrinol 180:155–162CrossRefGoogle Scholar
  46. Prevost N, Woulfs D, Tanaka T, Brass L (2002) Interactions between Eph kinases and ephrins provide a mechanism to support platelet aggregation once cell-to-cell contact has occurred. Proc Natl Acad Sci USA 99:9219–9224PubMedCrossRefGoogle Scholar
  47. Rico G, Díaz-Guerra O, Giménez-Scherer J, Kretschmer R (1992) Effect of the monocyte locomotion inhibitory factor (MLIF) produced by Entamoeba histolytica upon the respiratory burst of human leukocytes. Arch Med Res 23(2):157–159PubMedGoogle Scholar
  48. Rico G, Díaz-Guerra O, Kretschmer RR (1995) Cyclic nucleotide changes induced in human leukocytes by a product of axenically grown Entamoeba histolytica that inhibits human monocyte locomotion. Parasitol Res 81:158–162PubMedCrossRefGoogle Scholar
  49. Rico G, Leandro E, Rojas S, Giménez-Scherer J (2003) The monocyte locomotion inhibitory factor produced by Entamoeba histolytica inhibits induced nitric oxide production in human leukocytes. Parasitol Res 90(4):264–267PubMedCrossRefGoogle Scholar
  50. Robson MC, Hill DP, Smith PD, Wang X et al (2000) Sequential cytokine therapy for pressure ulcers: clinical and mechanistic response. Ann Surg 231:600–611PubMedCrossRefGoogle Scholar
  51. Sepúlveda B, Martínez-Palomo A (1982) Immunology of amoebiasis by Entamoeba histolytica. In: Cohen S, Warren KS (eds) Immunology of parasitic diseases. Blackwell, Oxford, pp 170–175Google Scholar
  52. Silva-García R, Rico G (2011) Anti-inflammatory defense mechanisms of Entamoeba histolytica. Inflamm Res 60(2):111–117PubMedCrossRefGoogle Scholar
  53. Sjolund M, Hedin U, Sejersen T, Heldin CH et al (1988) Arterial smooth muscle cells express platelet-derived growth factor (PDGF) A chain mRNA, secrete a PDGF-like mitogen, and bind exogenous PDGF in a phenotype- and growth state-dependent manner. J Cell Biol 106:403–413PubMedCrossRefGoogle Scholar
  54. Soldatova L, King R (2005) Are the current ontologies in biology good ontologies? Nat Biotechnol 23:1095–1098PubMedCrossRefGoogle Scholar
  55. Srivastava AS, Nedelcu E, Esmaeli-Azad B, Mishra R et al (2007) Thrombopoietin enhances generation of CD 34+ cells from human embryonic stem cells. Stem Cells 25(6):1456–1461PubMedCrossRefGoogle Scholar
  56. Sugiyama M, Ichida T, Sato T, Ishikawa T et al (1998) Expression of activin A is increased in cirrhotic and fibrotic rat livers. Gastroenterology 114:550–558PubMedCrossRefGoogle Scholar
  57. Suri C, Jones PF, Patan S, Bartunkova S et al (1996) Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell 87:1171–1180PubMedCrossRefGoogle Scholar
  58. Thompson TB, Lerch TF, Cook RW, Woodruff TK et al (2005) The structure of the follistatin: activin complex reveals antagonism of both type I and type II receptor binding. Dev Cell 9:535–543PubMedCrossRefGoogle Scholar
  59. Thurston G (2002) Complementary actions of VEGF and angiopoietin-1 on blood vessel growth and leakage. J Anat 200:575–580PubMedCrossRefGoogle Scholar
  60. Tremblay GM, Santrand MB, Jordana M, Gauldie J (1994) Fibroblasts as effectors cells in fibrosis. In: Phan SH, Thrall RS (eds) Pulmonary fibrosis. Marcel Dekker, New York, pp 541–577Google Scholar
  61. Utrera-Barillas D, Velazquez JR, Enciso A, Muñoz Cruz S, Rico G, Curiel Quezada E, Terán LM, Kretschmer R (2003) An anti-inflammatory oligopeptide produced by Entamoeba histolytica down-regulates the expression of pro-inflammatory chemokines. Parasite Immunol 25:475–482PubMedCrossRefGoogle Scholar
  62. Vale W, Hsurh A, Riviel C, Yu S (1995) Peptide growth factors and their receptors (handbook of experimental pharmacology). In: Sporn MA, Roberts AB (eds) The inhibin/activin family of growth factors. Springer, Heidelberg, pp 211–248Google Scholar
  63. Vancheri C, Gauldie J, Bienenstock J, Cox G et al (1989) Human lung fibroblast-derived granulocyte-macrophage colony stimulating factor (GM-CSF) mediates eosinophil survival in vitro. Am J Respir Cell Mol Biol 1:289–295PubMedGoogle Scholar
  64. Vancheri C, Mastruzzo C, Tomaselli V, Sortino MA et al (2001) Normal human lung fibroblasts differently modulate interleukin-10 and interleukin-12 production by monocytes: implications for an altered immune response in pulmonary chronic inflammation. Am J Respir Cell Mol Biol 25:592–599PubMedGoogle Scholar
  65. WHO (1995) The world health report 1995—bridging the gaps. World Health Forum 16:377–385Google Scholar
  66. Winning RS, Scales JB, Sargent TD (1996) Disruption of cell adhesion in Xenopus embryos by Pagliaccio an Eph-class receptor tyrosine kinase. Dev Biol 179:309–319PubMedCrossRefGoogle Scholar
  67. Yancopoulos GD, Davis S, Gale NW, Rudge JS et al (2000) Vascular-specific growth factors and blood vessel formation. Nature 407:242–248PubMedCrossRefGoogle Scholar
  68. Zamora DO, Babra B, Pan Y, Planck SR et al (2006) Human leukocytes express ephrinB2 which activates microvascular endothelial cells. Cell Immunol 242:99–109PubMedCrossRefGoogle Scholar
  69. Zisch AH, Stallcup WB, Chong LD, Dahlin-Huppe K et al (1997) Tyrosine phosphorylation of L1 family adhesion molecules: implication of the Eph kinase Cek5. J Neurosci Res 47:655–665PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • R. Silva-García
    • 1
  • M. E. Morales-Martínez
    • 1
  • F. Blanco-Favela
    • 1
  • A. Torres-Salazar
    • 2
  • J. Ríos-Olvera
    • 1
  • M. R. Garrido-Ortega
    • 1
  • E. Tesoro-Cruz
    • 1
    • 4
  • G. Rico-Rosillo
    • 3
  1. 1.Unidad de Investigación Médica en InmunologíaHospital de Pediatría, CMN-Siglo XXI IMSS. Av.MexicoMexico
  2. 2.Instituto de Biotecnología Universidad Autónoma de Nuevo LeónMonterreyMexico
  3. 3.División de Investigación, Facultad de Medicina, UNAM. Av.Universidad 3000 Circuito Escolar s/nMexicoMexico
  4. 4.Departamento de InvestigaciónExperimental y Bioterio del I.N.C.M.N “SZ”MexicoMexico

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