Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-23T04:12:31.497Z Has data issue: false hasContentIssue false
  • English
  • Français

Malaria pigment accelerates MTT – formazan exocytosis in human endothelial cells

Published online by Cambridge University Press:  01 October 2018

Sarah D'Alessandro ,
Yolanda Corbett Open the ORCID record for Yolanda Corbett [Opens in a new window] ,
Silvia Parapini ,
Federica Perego ,
Loredana Cavicchini ,
Lucia Signorini ,
Serena Delbue ,
Carla Perego ,
Pasquale Ferrante  and
Donatella Taramelli
...Show all authors
Sarah D'Alessandro
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
Yolanda Corbett
Affiliation:
Dipartimento di Bioscienze, Università degli Studi di Milano, Milano, Italy
Silvia Parapini
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
Federica Perego
Affiliation:
Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Milano, Italy
Loredana Cavicchini
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
Lucia Signorini
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
Serena Delbue
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
Carla Perego
Affiliation:
Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Milano, Italy
Pasquale Ferrante
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
Donatella Taramelli
Affiliation:
Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Milano, Italy
Nicoletta Basilico*
Affiliation:
Dipartimento di Scienze Biomediche, Chirurgiche e Odontoiatriche, Università degli Studi di Milano, Milano, Italy
*
Author for correspondence: Nicoletta Basilico, E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Haemozoin is a by-product of haemoglobin digestion by intraerythrocytic malaria parasites, which induces immunologic responses on different tissues, including endothelial cells. In the present paper, the incubation of human microvascular endothelial cells with haemozoin significantly inhibited MTT reduction, a measure of cytotoxicity, without increasing the release of cytoplasmic lactate dehydrogenase. Moreover, haemozoin did not induce apoptosis or cell cycle arrest nor decreased the number of live cells, suggesting that cells viability itself was not affected and that the inhibition of MTT reduction was only apparent and probably due to accelerated MTT-formazan exocytosis. After 30 min of MTT addition, a significant increase in the % of cells exocytosing MTT formazan crystals was observed in haemozoin-treated cells compared with control cells. Such an effect was partially reversed by the addition of genistein, an inhibitor of MTT-formazan exocytosis. The rapid release of CXCL-8, a preformed chemokine contained in Weibel–Palade bodies, confirmed that haemozoin induces a perturbation of the intracellular endothelial trafficking, including the exocytosis of MTT-formazan containing vesicles. The haem moiety of haemozoin is responsible for the observed effect. Moreover, this work underlines that MTT assay should not be used to measure cytotoxicity induced by haemozoin and other methods should be preferred.

Keywords

Endothelial cellshaemozoinmalariaMTT reduction
Type
Research Article
Information
Parasitology , Volume 146 , Issue 3 , March 2019 , pp. 399 - 406
Check for updates
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abe, K and Saito, H (1998) Amyloid beta protein inhibits cellular MTT reduction not by suppression of mitochondrial succinate dehydrogenase but by acceleration of MTT formazan exocytosis in cultured rat cortical astrocytes. Neuroscience Research 31, 295–305.Google Scholar
Ades, EW, Candal, FJ, Swerlick, RA, George, VG, Summers, S, Bosse, DC and Lawley, TJ (1992) HMEC-1: establishment of an immortalized human microvascular endothelial cell line. Journal of Investigative Dermatology 99, 683–690.Google Scholar
Andrews, KT and Lanzer, M (2002) Maternal malaria: Plasmodium falciparum sequestration in the placenta. Parasitology Research 88, 715–723.Google Scholar
Armah, HB, Wilson, NO, Sarfo, BY, Powell, MD, Bond, VC, Anderson, W, Adjei, AA, Gyasi, RK, Tettey, Y, Wiredu, EK, Tongren, JE, Udhayakumar, V and Stiles, JK (2007) Cerebrospinal fluid and serum biomarkers of cerebral malaria mortality in Ghanaian children. Malaria Journal 6, 147.Google Scholar
Ayimba, E, Hegewald, J, Ségbéna, AY, Gantin, RG, Lechner, CJ, Agosssou, A, Banla, M and Soboslay, PT (2011) Proinflammatory and regulatory cytokines and chemokines in infants with uncomplicated and severe Plasmodium falciparum malaria. Clinical and Experimental Immunology 166, 218–226.Google Scholar
Barrera, V, Skorokhod, OA, Baci, D, Gremo, G, Arese, P and Schwarzer, E (2011) Host fibrinogen stably bound to hemozoin rapidly activates monocytes via TLR-4 and CD11b/CD18-integrin: a new paradigm of hemozoin action. Blood 117, 5674–5682.Google Scholar
Basilico, N, Tognazioli, C, Picot, S, Ravagnani, F and Taramelli, D (2003) Synergistic and antagonistic interactions between haemozoin and bacterial endotoxin on human and mouse macrophages. Parassitologia 45, 135–140.Google Scholar
Basilico, N, Parapini, S, Sisto, F, Omodeo-Salè, F, Coghi, P, Ravagnani, F, Olliaro, P and Taramelli, D (2010) The lipid moiety of haemozoin (Malaria Pigment) and P. falciparum parasitised red blood cells bind synthetic and native endothelin-1. Journal of Biomedicine & Biotechnology 2010, 854927.Google Scholar
Basilico, N, Corbett, Y, D’ Alessandro, S, Parapini, S, Prato, M, Girelli, D, Misiano, P, Olliaro, P and Taramelli, D (2017) Malaria pigment stimulates chemokine production by human microvascular endothelium. Acta Tropica 172, 125–131.Google Scholar
Beeson, JG, Reeder, JC, Rogerson, SJ and Brown, GV (2001) Parasite adhesion and immune evasion in placental malaria. Trends in Parasitology 17, 331–337.Google Scholar
Berridge, MV and Tan, AS (1993) Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Archives of Biochemistry and Biophysics 303, 474–482.Google Scholar
Berridge, M, Tan, A, Mccoy, K and Wang, R (1996) The biochemical and cellular basis of cell proliferation assays that use tetrazolium salts. Biochemica 4, 14–16.Google Scholar
Berridge, MV, Herst, PM and Tan, AS (2005) Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnology Annual Review 11, 127–152.Google Scholar
Brown, H, Rogerson, S, Taylor, T, Tembo, M, Mwenechanya, J, Molyneux, M and Turner, G (2001) Blood-brain barrier function in cerebral malaria in Malawian children. American Journal of Tropical Medicine and Hygiene 64, 207–213.Google Scholar
Claus, V, Jahraus, A, Tjelle, T, Berg, T, Kirschke, H, Faulstich, H and Griffiths, G (1998) Lysosomal enzyme trafficking between phagosomes, endosomes, and lysosomes in J774 macrophages. Enrichment of cathepsin H in early endosomes. Journal of Biological Chemistry 273, 9842–9851.Google Scholar
D'Alessandro, S, Corbett, Y, Ilboudo, DP, Misiano, P, Dahiya, N, Abay, SM, Habluetzel, A, Grande, R, Gismondo, MR, Dechering, KJ, Koolen, KM, Sauerwein, RW, Taramelli, D, Basilico, N and Parapini, S (2015) Salinomycin and other ionophores as a new class of antimalarial drugs with transmission-blocking activity. Antimicrobial Agents and Chemotherapy 59, 5135–5144.Google Scholar
Deroost, K, Pham, TT, Opdenakker, G and Van den Steen, PE (2016) The immunological balance between host and parasite in malaria. FEMS Microbiology Reviews 40, 208–257.Google Scholar
Diaz, G, Melis, M, Musin, A, Piludu, M, Piras, M and Falchi, AM (2007) Localization of MTT formazan in lipid droplets. An alternative hypothesis about the nature of formazan granules and aggregates. European Journal of Histochemistry 51, 213–218.Google Scholar
Egan, TJ (2008) Haemozoin formation. Molecular and Biochemical Parasitology 157, 127–136.Google Scholar
Fisichella, M, Dabboue, H, Bhattacharyya, S, Saboungi, ML, Salvetat, JP, Hevor, T and Guerin, M (2009) Mesoporous silica nanoparticles enhance MTT formazan exocytosis in HeLa cells and astrocytes. Toxicology in Vitro 23, 697–703.Google Scholar
Giribaldi, G, Ulliers, D, Schwarzer, E, Roberts, I, Piacibello, W and Arese, P (2004) Hemozoin- and 4-hydroxynonenal-mediated inhibition of erythropoiesis. Possible role in malarial dyserythropoiesis and anemia. Haematologica 89, 492–493.Google Scholar
Goldie, P, Roth, EF, Oppenheim, J and Vanderberg, JP (1990) Biochemical characterization of Plasmodium falciparum hemozoin. American Journal of Tropical Medicine and Hygiene 43, 584–596.Google Scholar
Guiguemde, WA, Hunt, NH, Guo, J, Marciano, A, Haynes, RK, Clark, J, Guy, RK and Golenser, J (2014) Treatment of murine cerebral malaria by artemisone in combination with conventional antimalarial drugs: antiplasmodial effects and immune responses. Antimicrobial Agents and Chemotherapy 58, 4745–4754.Google Scholar
Hollestelle, MJ, Donkor, C, Mantey, EA, Chakravorty, SJ, Craig, A, Akoto, AO, O'Donnell, J, van Mourik, JA and Bunn, J (2006) von Willebrand factor propeptide in malaria: evidence of acute endothelial cell activation. British Journal of Haematology 133, 562–569.Google Scholar
Into, T, Kanno, Y, Dohkan, J, Nakashima, M, Inomata, M, Shibata, K, Lowenstein, CJ and Matsushita, K (2007) Pathogen recognition by Toll-like receptor 2 activates Weibel-Palade body exocytosis in human aortic endothelial cells. Journal of Biological Chemistry 282, 8134–8141.Google Scholar
Isobe, I, Michikawa, M and Yanagisawa, K (1999) Enhancement of MTT, a tetrazolium salt, exocytosis by amyloid beta-protein and chloroquine in cultured rat astrocytes. Neuroscience Letters 266, 129–132.Google Scholar
Jaramillo, M, Godbout, M and Olivier, M (2005) Hemozoin induces macrophage chemokine expression through oxidative stress-dependent and -independent mechanisms. Journal of Immunology 174, 475–484.Google Scholar
Khadjavi, A, Giribaldi, G and Prato, M (2010) From control to eradication of malaria: the end of being stuck in second gear? Asian Pacific Journal of Tropical Medicine 3, 412–420.Google Scholar
Khaw, LT, Ball, HJ, Golenser, J, Combes, V, Grau, GE, Wheway, J, Mitchell, AJ and Hunt, NH (2013) Endothelial cells potentiate interferon-γ production in a novel tripartite culture model of human cerebral malaria. PLoS ONE 8, e69521.Google Scholar
Lamikanra, AA, Theron, M, Kooij, TW and Roberts, DJ (2009) Hemozoin (malarial pigment) directly promotes apoptosis of erythroid precursors. PLoS ONE 4, e8446.Google Scholar
Liu, Y and Schubert, D (1997) Cytotoxic amyloid peptides inhibit cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction by enhancing MTT formazan exocytosis. Journal of Neurochemistry 69, 2285–2293.Google Scholar
Liu, Y and Schubert, D (1998) Steroid hormones block amyloid fibril-induced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) formazan exocytosis: relationship to neurotoxicity. Journal of Neurochemistry 71, 2322–2329.Google Scholar
Liu, Y, Peterson, DA, Kimura, H and Schubert, D (1997) Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. Journal of Neurochemistry 69, 581–593.Google Scholar
Liu, Y, Peterson, DA and Schubert, D (1998) Amyloid beta peptide alters intracellular vesicle trafficking and cholesterol homeostasis. Proceedings of the National Academy of Sciences of the United States of America 95, 13266–13271.Google Scholar
Liu, M, Dickinson-Copeland, C, Hassana, S and Stiles, JK (2016) Plasmodium-infected erythrocytes (pRBC) induce endothelial cell apoptosis via a heme-mediated signaling pathway. Drug Design, Development and Therapy 10, 1009–1018.Google Scholar
Maknitikul, S, Luplertlop, N, Chaisri, U, Maneerat, Y and Ampawong, S (2018) Featured article: immunomodulatory effect of hemozoin on pneumocyte apoptosis via CARD9 pathway, a possibly retarding pulmonary resolution. Experimental Biology and Medicine (Maywood) 243, 395–407.Google Scholar
Matsushita, K, Yamakuchi, M, Morrell, CN, Ozaki, M, O'Rourke, B, Irani, K and Lowenstein, CJ (2005) Vascular endothelial growth factor regulation of Weibel-Palade-body exocytosis. Blood 105, 207–214.Google Scholar
Metcalf, DJ, Nightingale, TD, Zenner, HL, Lui-Roberts, WW and Cutler, DF (2008) Formation and function of Weibel-Palade bodies. Journal of Cell Science 121, 19–27.Google Scholar
Molinari, BL, Tasat, DR, Palmieri, MA and Cabrini, RL (2005) Kinetics of MTT-formazan exocytosis in phagocytic and non-phagocytic cells. Micron 36, 177–183.Google Scholar
Mosmann, T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65, 55–63.Google Scholar
Omodeo-Salè, F, Basilico, N, Folini, M, Olliaro, P and Taramelli, D (1998) Macrophage populations of different origins have distinct susceptibilities to lipid peroxidation induced by beta-haematin (malaria pigment). FEBS Letters 433, 215–218.Google Scholar
Omodeo-Sale, F, Motti, A, Basilico, N, Parapini, S, Olliaro, P and Taramelli, D (2003) Accelerated senescence of human erythrocytes cultured with Plasmodium falciparum. Blood 102, 705–711.Google Scholar
Pagola, S, Stephens, PW, Bohle, DS, Kosar, AD and Madsen, SK (2000) The structure of malaria pigment beta-haematin. Nature 404, 307–310.Google Scholar
Parroche, P, Lauw, FN, Goutagny, N, Latz, E, Monks, BG, Visintin, A, Halmen, KA, Lamphier, M, Olivier, M, Bartholomeu, DC, Gazzinelli, RT and Golenbock, DT (2007) Malaria hemozoin is immunologically inert but radically enhances innate responses by presenting malaria DNA to Toll-like receptor 9. Proceedings of the National Academy of Sciences of the United States of America 104, 1919–1924.Google Scholar
Polimeni, M, Valente, E, Aldieri, E, Khadjavi, A, Giribaldi, G and Prato, M (2013) Role of 15-hydroxyeicosatetraenoic acid in hemozoin-induced lysozyme release from human adherent monocytes. Biofactors 39, 304–314.Google Scholar
Pongponratn, E, Turner, GD, Day, NP, Phu, NH, Simpson, JA, Stepniewska, K, Mai, NT, Viriyavejakul, P, Looareesuwan, S, Hien, TT, Ferguson, DJ and White, NJ (2003) An ultrastructural study of the brain in fatal Plasmodium falciparum malaria. American Journal of Tropical Medicine and Hygiene 69, 345–359.Google Scholar
Prato, M and Giribaldi, G (2011) Matrix metalloproteinase-9 and haemozoin: wedding rings for human host and Plasmodium falciparum parasite in complicated malaria. Journal of Tropical Medicine 2011, ID 628435.Google Scholar
Prato, M, Gallo, V, Giribaldi, G and Arese, P (2008) Phagocytosis of haemozoin (malarial pigment) enhances metalloproteinase-9 activity in human adherent monocytes: role of IL-1beta and 15-HETE. Malaria Journal 7, 157.Google Scholar
Prato, M, D'Alessandro, S, Van den Steen, PE, Opdenakker, G, Aresel, P, Taramelli, D and Basilico, N (2011) Modulation of Matrix Metalloproteinases and their tissue inhibitors in haemozoin-treated human microvascular endothelial cells. Febs Journal 278, 310–310.Google Scholar
Schwarzer, E, Kuhn, H, Valente, E and Arese, P (2003) Malaria-parasitized erythrocytes and hemozoin nonenzymatically generate large amounts of hydroxy fatty acids that inhibit monocyte functions. Blood 101, 722–728.Google Scholar
Shio, MT, Kassa, FA, Bellemare, MJ and Olivier, M (2010) Innate inflammatory response to the malarial pigment hemozoin. Microbes and Infection 12, 889–899.Google Scholar
Skorokhod, OA, Caione, L, Marrocco, T, Migliardi, G, Barrera, V, Arese, P, Piacibello, W and Schwarzer, E (2010) Inhibition of erythropoiesis in malaria anemia: role of hemozoin and hemozoin-generated 4-hydroxynonenal. Blood 116, 4328–4337.Google Scholar
Stockert, JC, Blázquez-Castro, A, Cañete, M, Horobin, RW and Villanueva, A (2012) MTT assay for cell viability: intracellular localization of the formazan product is in lipid droplets. Acta Histochemica 114, 785–796.Google Scholar
Taramelli, D, Basilico, N, De Palma, AM, Saresella, M, Ferrante, P, Mussoni, L and Olliaro, P (1998) The effect of synthetic malaria pigment (beta-haematin) on adhesion molecule expression and interleukin-6 production by human endothelial cells. Transactions of the Royal Society of Tropical Medicine and Hygiene 92, 57–62.Google Scholar
Taramelli, D, Recalcati, S, Basilico, N, Olliaro, P and Cairo, G (2000) Macrophage preconditioning with synthetic malaria pigment reduces cytokine production via heme iron-dependent oxidative stress. Laboratory Investigation 80, 1781–1788.Google Scholar
Traganos, F and Darzynkiewicz, Z (1994) Lysosomal proton pump activity: supravital cell staining with acridine orange differentiates leukocyte subpopulations. Methods in Cell Biology 41, 185–194.Google Scholar
Tyberghein, A, Deroost, K, Schwarzer, E, Arese, P and Van den Steen, PE (2014) Immunopathological effects of malaria pigment or hemozoin and other crystals. Biofactors 40, 59–78.Google Scholar
Utgaard, JO, Jahnsen, FL, Bakka, A, Brandtzaeg, P and Haraldsen, G (1998) Rapid secretion of prestored interleukin 8 from Weibel-Palade bodies of microvascular endothelial cells. Journal of Experimental Medicine 188, 1751–1756.Google Scholar
Vischer, UM and Wagner, DD (1993) CD63 is a component of Weibel-Palade bodies of human endothelial cells. Blood 82, 1184–1191.Google Scholar
Yeo, TW, Lampah, DA, Gitawati, R, Tjitra, E, Kenangalem, E, Piera, K, Price, RN, Duffull, SB, Celermajer, DS and Anstey, NM (2008) Angiopoietin-2 is associated with decreased endothelial nitric oxide and poor clinical outcome in severe falciparum malaria. Proceedings of the National Academy of Sciences of the United States of America 105, 17097–17102.Google Scholar