Role of glutathione, ROS, and Bcl-xL in the inhibition of apoptosis of monocyte-derived dendritic cells by Leishmania mexicana promastigotes
- 120 Downloads
Dendritic cells (DCs) are one of the principal host cells of the obligate intracellular parasite Leishmania that can survive and reproduce within cells due to the ability to regulate different cellular events, including apoptosis. Inhibition of host cell apoptosis is a strategy employed by multiple pathogens to ensure their survival in the infected cell. We have previously reported that Leishmania mexicana promastigotes and amastigotes inhibit camptothecin-induced apoptosis of monocyte-derived dendritic cells (moDCs) through the downregulation of p38 and JNK phosphorylation. The upregulation of glutathione (GSH), the most important regulator of reactive oxygen species (ROS) concentration, has proven to protect cells from apoptosis through the inhibition of JNK1. Another mechanism employed by cells for the protection of apoptosis is the expression of anti-apoptotic proteins of the Bcl-2 family. The aim of this study was to determine if GSH, ROS, and Bcl-xL participate in the inhibition of camptothecin-induced apoptosis of moDC by L. mexicana promastigotes. GSH quantification assays showed that camptothecin and BSO (an inhibitor of glutathione synthesis) strongly decreased intracellular GSH concentration in moDC, while infection with L. mexicana promastigotes had no effect in the level of GSH. On the other hand, infection with L. mexicana promastigotes of BSO- and camptothecin-treated moDC diminished the concentration of ROS and induced the expression of the anti-apoptotic protein Bcl-xL. Our findings suggest that inhibition of camptothecin-induced apoptosis of moDC by L. mexicana promastigotes is preferentially regulated by the expression of anti-apoptotic proteins of the Bcl-2 family rather than by the redox status of the cell.
KeywordsApoptosis Bcl-xL Dendritic cells Glutathione Leishmania mexicana
We thank Dr. Alfonso Olivos García for his valuable help with the ROS determinations.
This work was supported by grant IN225116 from Papiit, DGAPA-UNAM. Jorge Rodríguez-González is a Ph. D student of Posgrado en Ciencias Biológicas, Facultad de Medicina, Universidad Nacional Autónoma de México and is recipient of a CONACyT fellowship, number 481304.
Compliance with ethical standards
Informed consent was obtained for the use of blood samples according to the Declaration of Helsinki and the Comisiones de Investigación y Ética de la Facultad de Medicina, Universidad Nacional Autónoma de México approved the protocol with number 104/2015 on August 25, 2015 with a validity of 3 years.
- Akarid K, Arnoult D, Micic-Polianski J, Sif J, Estaquier J, Ameisen JC (2004) Leishmania major-mediated prevention of programmed cell death induction in infected macrophages is associated with the repression of mitochondrial release of cytochrome c. J Leukoc Biol 76(1):95–103. https://doi.org/10.1189/jlb.1001877 CrossRefPubMedGoogle Scholar
- Armstrong JS, Steinauer KK, Hornung B, Irish JM, Lecane P, Birrell GW, Peehl DM, Knox SJ (2002) Role of glutathione depletion and reactive oxygen species generation in apoptotic signaling in a human B lymphoma cell line. Cell Death Differ 9(3):252–263. https://doi.org/10.1038/sj.cdd.4400959 CrossRefPubMedGoogle Scholar
- Armstrong JS, Whiteman M, Yang H, Jones DP, Sternberg P Jr (2004) Cysteine starvation activates the redox-dependent mitochondrial permeability transition in retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 45(11):4183–4189. https://doi.org/10.1167/iovs.04-0570 CrossRefPubMedGoogle Scholar
- Carneiro PP, Conceicao J, Macedo M, Magalhaes V, Carvalho EM, Bacellar O (2016) The role of nitric oxide and reactive oxygen species in the killing of Leishmania braziliensis by monocytes from patients with cutaneous leishmaniasis. PLoS One 11(2):e0148084. https://doi.org/10.1371/journal.pone.0148084 CrossRefPubMedPubMedCentralGoogle Scholar
- Cruz KK, Fonseca SG, Monteiro MC, Silva OS, Andrade VM, Cunha FQ, Romão PR (2008) The influence of glutathione modulators on the course of Leishmania major infection in susceptible and resistant mice. Parasite Immunol 30(3):171–174. https://doi.org/10.1111/j.1365-3024.2007.01014.x CrossRefPubMedGoogle Scholar
- Dalton TP, Dieter MZ, Yang Y, Shertzer HG, Nebert DW (2000) Knockout of the mouse glutamate cysteine ligase catalytic subunit (Gclc) gene: embryonic lethal when homozygous, and proposed model for moderate glutathione deficiency when heterozygous. Biochem Biophys Res Comm 279(2):324–329. https://doi.org/10.1006/bbrc.2000.3930 CrossRefPubMedGoogle Scholar
- Galadari S, Rahman A, Pallichankandy S, Thayyullathil F (2017) Reactive oxygen species and cancer paradox: to promote or to suppress? Free Rad Biol Med 104:144–164. https://doi.org/10.1016/j.freeradbiomed.2017.01.004 CrossRefPubMedGoogle Scholar
- Gupta P, Srivastav S, Saha S, Das PK, Ukil A (2016) Leishmania donovani inhibits macrophage apoptosis and pro-inflammatory response through AKT-mediated regulation of beta-catenin and FOXO-1. Cell Death Differ 23(11):1815–1826. https://doi.org/10.1038/cdd.2016.101 CrossRefPubMedPubMedCentralGoogle Scholar
- Handley ME, Thakker M, Pollara G, Chain BM, Katz DR (2005) JNK activation limits dendritic cell maturation in response to reactive oxygen species by the induction of apoptosis. Free Rad Biol Med 38(12):1637–1652. https://doi.org/10.1016/j.freeradbiomed.2005.02.022 CrossRefPubMedGoogle Scholar
- Lisi S, Sisto M, Acquafredda A, Spinelli R, Schiavone M, Mitolo V, Brandonisio O, Panaro M (2005) Infection with Leishmania infantum inhibits actinomycin D-induced apoptosis of human monocytic cell line U-937. J Euk Microbiol 52(3):211–217. https://doi.org/10.1111/j.1550-7408.2005.00026.x CrossRefPubMedGoogle Scholar
- Mantzaris MD, Bellou S, Skiada V, Kitsati N, Fotsis T, Galaris D (2016) Intracellular labile iron determines H2O2-induced apoptotic signaling via sustained activation of ASK1/JNK-p38 axis. Free Rad Biol Med 97:454–465. https://doi.org/10.1016/j.freeradbiomed.2016.07.002 CrossRefPubMedGoogle Scholar
- Rodriguez-Gonzalez J, Wilkins-Rodriguez A, Argueta-Donohue J, Gutierrez-Kobeh L (2016) Leishmania mexicana promastigotes down regulate JNK and p-38 MAPK activation: Role in the inhibition of camptothecin-induced apoptosis of monocyte-derived dendritic cells. Exp Parasitol 163:57–67. https://doi.org/10.1016/j.exppara.2015.12.005 CrossRefPubMedGoogle Scholar
- Rogers M, Kropf P, Choi BS, Dillon R, Podinovskaia M, Bates P, Müller I (2009) Proteophosophoglycans regurgitated by Leishmania-infected sand flies target the L-arginine metabolism of host macrophages to promote parasite survival. PLoS Pathog 5(8):e1000555. https://doi.org/10.1371/journal.ppat.1000555 CrossRefPubMedPubMedCentralGoogle Scholar
- Sarkar A, Aga E, Bussmeyer U, Bhattacharyya A, Möller S, Hellberg L, Behnen M, Solbach W, Laskay T (2013) Infection of neutrophil granulocytes with Leishmania major activates ERK 1/2 and modulates multiple apoptotic pathways to inhibit apoptosis. Med Microbiol Immunol 202(1):25–35. https://doi.org/10.1007/s00430-012-0246-1 CrossRefPubMedGoogle Scholar
- Son Y, Kim S, Chung HT, Pae HO (2013) Reactive oxygen species in the activation of MAP kinases. Methods Enzymol 528:27–48. https://doi.org/10.1016/B978-0-12-405881-1.00002-1 CrossRefPubMedGoogle Scholar
- Srivastav S, Basu Ball W, Gupta P, Giri J, Ukil A, Das PK (2014) Leishmania donovani prevents oxidative burst-mediated apoptosis of host macrophages through selective induction of suppressors of cytokine signaling (SOCS) proteins. J Biol Chem 289(2):1092–1105. https://doi.org/10.1074/jbc.M113.496323 CrossRefPubMedGoogle Scholar
- Valdes-Reyes L, Argueta J, Morán J, Salaiza N, Hernández J, Berzunza M, Becker I, Gutiérrez-Kobeh L (2009) Leishmania mexicana: inhibition of camptothecin-induced apoptosis of monocyte-derived dendritic cells. Exp Parasitol 121(3):199–207. https://doi.org/10.1016/j.exppara.2008.10.020 CrossRefPubMedGoogle Scholar
- Van Laethem A, Nys K, Van Kelst S, Claerhout S, Ichijo H, Vandenheede JR, Garmyn M, Agostinis P (2006) Apoptosis signal regulating kinase-1 connects reactive oxygen species to p38 MAPK-induced mitochondrial apoptosis in UVB-irradiated human keratinocytes. Free Radic Biol Med 41(9):1361–1371. https://doi.org/10.1016/j.freeradbiomed.2006.07.007 CrossRefPubMedGoogle Scholar
- Vazquez-Lopez R, Argueta-Donohue J, Wilkins-Rodriguez A, Escalona-Montano A, Gutierrez-Kobeh L (2015) Leishmania mexicana amastigotes inhibit p38 and JNK and activate PI3K/AKT: role in the inhibition of apoptosis of dendritic cells. Parasite Immunol 37(11):579–589. https://doi.org/10.1111/pim.12275 CrossRefPubMedGoogle Scholar
- Zimmermann AK, Loucks FA, Schroeder EK, Bouchard RJ, Tyler KL, Linseman DA (2007) Glutathione binding to the Bcl-2 homology-3 domain groove: a molecular basis for Bcl-2 antioxidant function at mitochondria. J Biol Chem 282(40):29296–29304. https://doi.org/10.1074/jbc.M702853200 CrossRefPubMedPubMedCentralGoogle Scholar