Effect of the Monocyte Locomotion Inhibitory Factor (MLIF) a Natural Anti-Inflammatory Produced by E. Histolytica on Apoptosis in Human CD4 + T Lymphocytes
Sara Rojas-Dotor, Liliana Vargas-Neri, Francisco Blanco-Favela
DOI: 10.4236/pp.2011.24032   PDF    HTML     5,611 Downloads   10,583 Views   Citations


Apoptosis prevents the extravasation of intracellular material and the subsequent inflammatory response. Currently, it is not known whether Monocyte Locomotion Inhibitor Factor (MLIF), an anti-inflammatory pentapeptide, induces programmed cell death. We evaluated the effect of MLIF on extrinsic and intrinsic apoptosis pathways human CD4 + T lymphocytes. Cells were cultured for 24 h in RPMI-1640 medium alone (control) or in RPMI medium containing MLIF alone, PMA alone, PMA + MLIF or actinomycin D. Annexin V/propidium iodide-stained cells in early apoptosis showed that cells treated with MLIF or PMA + MLIF were not significantly different from control cells in medium; in contrast, cells treated with PMA or PMA + MLIF demonstrated significant differences from the control in delayed apoptosis. Cytochrome c and caspase 3 levels in cells treated with MLIF showed no significant differences from control cells, however, compared to the control, cells treated with PMA and PMA + MLIF exhibited a significant increase in cytochrome c and caspase 3 levels, which demonstrates that this weak induction of cell death is regulated by the intrinsic pathway of apoptosis. The Fas receptor was not detected in cell culture with any of the treatments employed, suggesting that the extrinsic pathway of apoptosis is not involved. The MLIF per se does not induce apoptosis in human CD4 + T lymphocytes; there may be an additional effect of PMA + MLIF producing the low levels of cell death recorded in the late apoptosis phase. MLIF acts as a natural, biological anti-inflammatory compound produced in axenic cultures of E. histolytica that does not cause apoptosis or elicit an immune response due to its small size, which could make it a strong candidate for future clinical applications.

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S. Rojas-Dotor, L. Vargas-Neri and F. Blanco-Favela, "Effect of the Monocyte Locomotion Inhibitory Factor (MLIF) a Natural Anti-Inflammatory Produced by E. Histolytica on Apoptosis in Human CD4 + T Lymphocytes," Pharmacology & Pharmacy, Vol. 2 No. 4, 2011, pp. 248-255. doi: 10.4236/pp.2011.24032.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] P. Marrack, T. Mitchell, J. Bender, D. Hildeman, R. Kedl, K. Teague and J. Kappler, “T-cell survival,” Immunol Rev, Vol. 165, 1998, pp 279-285. doi:10.1111/j.1600-065X.1998.tb01245.x
[2] J. P. Medema, “Apoptosis-life and death in a FLASH,” Nature, Vol.398, No. 5, 1999, pp 756-757. doi:10.1038/19638
[3] C. Klas, K. M. Debatin, R. R. Jonker and P. H. Krammer, “Activation interferes with the APO-1 pathway in mature human T cells,” Int Immunol, Vol. 5, No. 6, 1993, pp 625-630. doi:10.1093/intimm/5.6.625
[4] G. M. Bran, J. Stern-Strater, k. H?rmann, F. Riedel and U. R. Goessler, “Apoptosis in bone for tissue engineering,” Arch Med Res, Vol.39, No.5, 2008, pp 467-482. doi:10.1016/j.arcmed.2008.02.007
[5] D. J. Van Antwerp, S. J. T. Martin, Kafri, D. R. Green, and I. M. Verma, “Suppression of TNF-alpha-induced apoptosis by NF-kB,” Science, Vol. 274, No. 5288, 1996, pp 787-789. doi:10.1126/science.274.5288.787
[6] C. Scaffidi, S. Fulda, A. Srinivasan, C. Friesen, F. Li, K. J. Tomaselli, K. M. Debatin, P.H. Krammer and M. E. Peter, “Two CD95 (APO-1/Fas) signaling pathways,” EMBO J, Vol. 17, No. 6, 1998, pp 1675-1687. doi:10.1093/emboj/17.6.1675
[7] D. R. Schultz and W. J. Harrington, “Apoptosis: programmed cell death at a molecular level,” Semin Arthritis Rheum, Vol.32, No. 6, 2003, pp 345-369. doi:10.1053/sarh.2003.50005
[8] Y. S. Ki, E. Y. Park, H. W. Lee, M. S. Oh, Y. W. Cho, Y. K. Kwon, J. H. Moon and K. T. Lee “Donepezil, a potent acetylcholinesterase inhibitor, induces caspase-dependent apoptosis in human promyelocytic leukemia HL-60 cells,” Biol Pharm Bull, Vol. 33, No.6, 2010, pp. 1054- 1059. doi:10.1248/bpb.33.1054
[9] A. Lawen, “Apoptosis-an introduction 2,” Bioassay, Vol. 25, No. 9, 2003, pp 2013-2026. doi:10.1002/bies.10329
[10] K. W. Tinsley, D. Herzog and D. Leitenberg, “CD4 co-receptor dependent signaling promotes competency for re-stimulation induced cell death of effector T cells,” Cell Immunol, Vol. 266, No. 2, 2011, pp 200-2007. doi:10.1016/j.cellimm.2010.10.006
[11] T. Miyawaki, T. Uehara, R. Nibu, T. Tsuji, A. Yachie, S. Yonehara and N. Taniguchi “Differential expression of apoptosis-related Fas antigen on lymphocyte subpopulations in human peripheral blood,” J Immunol, Vol. 149, No. 11, 1992, pp 3753-3758.
[12] R. R. Kretschmer, M. l. Collado, M. Pacheco, M. Salinas, M. Lopéz-Osuna, M. Lecuona, E. Castro and J. Arellano, “Inhibition of human monocyte locomotion by products of axenically grown E. histolytica,” Parasite Immunol, Vol. 7, No. 5, 1985, pp 527-543. doi:10.1111/j.1365-3024.1985.tb00097.x
[13] G. Rico, O. Diaz-Guerra, J. A. Giménez-Scherer and R. R. Kretschmer, “Effect of the monocyte locomotion inhibitory factor (MLIF) produced by Entamoeba histolytica upon the respiratory burst of human leukocytes,” Arch Med Res, Vol. 23, No. 2, 1992, pp 157-159.
[14] G. Rico, E. Leandro, S. Rojas, J. A. Giménez, and R. R. Kretschmer, “The monocyte locomotion inhibitory factor produced by Entamoeba histolytica inhibits induced nitric oxide production in human leukocytes,” Parasitol Res, Vol. 90, No. 4, 2003, pp 264-267. doi:10.1007/s00436-002-0780-7
[15] J. A. Giménez-Scherer, G. Rico-Rosillo, J. Fernández- Díez and R. R. Kretschmer, “Inhibition of contact cutaneous delayed hypersensitivity reactions to DNBC in guinea pigs by the monocyte locomotion inhibitory factor (MLIF) produced by axenically grown Entamoeba histolytica,” Arch Med Res, Vol. 28, No. 4, 1997, pp. 237-238.
[16] J. A. Giménez-Scherer, G. Cárdenas, M. López-Osuna, J. R. Velázquez, G. Rico, A. Isibasi, M. C. Maldonando, M. E. Morales, J. Fernandez-Díez J and R.R. Kretschmer, “Immunization with a tetramer derivative of an anti-inflammatory pentapeptide produced by Entamoeba histolytica protects gerbils (Meriones unguiculatus) against experimental amoebic abscess of the liver,” Parasite Immunol, Vol. 26, No. 8, 2004, pp. 343-349. doi:10.1111/j.0141-9838.2004.00718.x
[17] C. Soriano-Correa, J. F. Sanchez-Ruiz, G. Rico-Rosillo, J. A. Giménez-Scherer, J.R. Velázquez, and R. Kretschmer, “Electronic structure and physicochemical properties of the anti-inflammatory pentapeptide produced by Entamoeba histolytica: Theoretical study,” J Molecular Structures: THEOCHEM, Vol.769, No. 91, 2006, pp 91-95. http://dx.doi.org/10.1016/j.theochem.2006.02.031
[18] A. B?yum, “Isolation of mononuclear cells and granulocytes from human blood,” Scand J Clin Lab In vest, Vol. 97, No. 97, 1968, pp 77-89.
[19] R. Freshney, “Scaling-up of animal cell cultures”, In: Rickwood D, Hames BD, Culture of Animal Cells: A Manual of Basic Technique. New York, 1987, pp 47-92.
[20] D. R. Green, “Apoptotic pathways: the roads to ruin,” Cell, vol. 94, No. 6, 1998, pp 695-698. doi.10.1016/S0092-8674(00)81728-6
[21] K. Chadee and E. Meerovitch, “The pathogenesis of experimentally induced amebic liver abscess in the gerbil (Meriones unguiculatus),” Am J pathol, Vol. 117, No.1, 1984, pp 71-80.
[22] S. Rojas-Dotor, G. Rico, J. Pérez, J. Velázquez and R. Kretschmer, “Cytokine expression in CD4+ cells exposed to the monocyte locomotion inhibitory factor produced by Entamoeba histolytica,” Parasitol Res, Vol. 98, No.5, 2006, pp 493-495. doi.10.1007/s00436-005-0090-y
[23] S. Rojas-Dotor, J. Pérez-Ramos, J. A. Giménez-Scherer, F. Blanco-Favela and G. Rico-Rosillo, “Effect of the monocyte locomotion inhibitory factor (MLIF) produced by E. histolityca on cytokines and chemokine receptors in T CD4+ lymphocytes,” Biol Res, Vol. 42, No.4, 2009, pp 415-425.
[24] S. Yonehara, A. Ishii and M. Yonehara, “A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor,” J Exp Med, Vol. 169, No. 5, 1989, pp. 1747-1756. doi:10.1084/jem.169.5.1747
[25] B. C. Trauth, A. M. Peters, S. Matzku, P. M?ller, W. Falk, K. M. Debatin and P. H. Krammer, “Monoclonal antibody-mediated tumor regression by induction of apoptosis,” Science, Vol. 245, No. 4915, 1989, pp 301-305. doi:10.1126/science.2787530
[26] G. A. Porter and R. J?nicke, “Emerging roles of caspase-3 in apoptosis,” Cell Death and Differentiation”, Vol. 6, No. 2, 1999, pp 99-104. doi:10.1038/sj.cdd.4400476
[27] J. P. Medema and J. Borst, “T cell signaling: a decision of life and death,” Hum Immunol, Vol. 60, No. 5, 1999, pp 403-11. doi:10.1016/S0198-8859(99)00008-7
[28] E. O’Flaherty, W. K. Wong, S. J. Pettit, K. Seymour, S. Ali and J.A. Kirby, “Regulation of T-cell apoptosis: a mixed lymphocyte reaction model,” Immunology, Vol. 100, No. 3, 2000 pp 289-99. doi:10.1046/j.1365-2567.2000.00048.x
[29] R. R. Kretschmer, E. M. Castro, G. Pacheco, G. Rico, O. Diaz-Guerra and J. Arellano, “The role of mannose in the receptor of the monocyte locomotion inhibitory factor produced by Entamoeba histolytica,” Parasitol Res, Vol. 77, No.5, 1991, pp 374-378. doi:10.1007/BF00931631
[30] G. Rico, O. Díaz-Guerra and R. Kretschmer, “Cyclic nucleotide changes induced in human leukocytes by a product of axenically grown Entamoeba histolytica that inhibits human monocyte locomotion,” Parasitol Res, Vol. 81, No. 5, 1995, pp 158-162. doi:10.1007/BF00931623
[31] D. Utrera-Barrillas, J. R. Velázquez, A. Enciso, S. M. Cruz, G. Rico, E. Curiel-Quezada, L. M. Terán and R. Kretschmer, “An anti-inflammatory oligopeptide produced by Entamoeba histolytica down-regulates the expression of pro-inflammatory chemokines,” Parasite Immunol, Vol. 25, No. 10, 2003, pp 475-482. doi:10.1111/j.1365-3024.2003.00657.x
[32] R. Kretschmer, J. Velázquez, D. Utreras-Barillas and A. Zentella, “The amebic anti nflammatory monocyte locomotion MLIF inhibits the NF kB nuclear translocation in human monocytes,” FASEB J, Vol. 18, No. 8, 2004, pp A1147.

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