Original Article

Induction of Apoptosis in Toxoplasma gondii Infected Hela Cells by Cisplatin and Sodium Azide and Isolation of Apoptotic Bodies and Potential Use for Vaccination against Toxoplasma gondii

Abstract

Background: Toxoplasma gondii can infect a wide range of mammalians, especially humans. It controls several intracellular signals for the inhibition of apoptosis. This study aimed to investigate the apoptogenic effect of cisplatin and sodium azide on T. gondii infected HeLa cells and isolate apoptotic bodies (blebs) as a potent stimulator of the immune system.

Methods: The cytotoxic properties of cisplatin and sodium azide (NaN3) on HeLa cells were evaluated by MTT assay. Moreover, the apoptogenic activity of cisplatin and NaN3 was studied using flow cytometry (Annexin V/PI double staining) and scanning electron microscopy (SEM). Finally, apoptotic bodies were separated by centrifugation.

Results: MTT assay data showed that the survival rate of cells treated with different concentration of NaN3 was significantly reduced, compared to negative control groups. Concerning cisplatin, only concentration of 20 μM had not a significant impact on the cell viability; however, the other concentration of cisplatin significantly reduced cell viability, compared to negative control groups. The level of early apoptosis in uninfected HeLa cells was higher compared to infected HeLa cells treated with cisplatin and NaN3. Finally, apoptotic bodies were separated from T. gondii infected HeLa cells treated with cisplatin. Conclusion: Apoptosis was induced in both uninfected and infected HeLa cells with T. gondii and apoptotic bodies were isolated from infected cells. Therefore, further studies on apoptotic bodies are required in order to find a proper candidate for vaccine preparation against T. gondii infections.

Webster JP. Dubey, JP Toxoplasmosis of Animals and Humans. BioMed Central; 2010.

Derouin F, Pelloux H. Prevention of toxoplasmosis in transplant patients. Clinical Microbiology and Infection. 2008;14(12):1089-101.

Awan K. Congenital toxoplasmosis: chances of occurrence in subsequent siblings. Annals of ophthalmology. 1978;10(4):459-65.

Kodjikian L, Hoigne I, Adam O, Jacquier P, Aebi-Ochsner C, Aebi C, et al. Vertical transmission of toxoplasmosis from a chronically infected immunocompetent woman. The Pediatric infectious disease journal. 2004;23(3):272-4.

Vogel N, Kirisits M, Michael E, Bach H, Hostetter M, Boyer K, et al. Congenital toxoplasmosis transmitted from an immunologically competent mother infected before conception. Clinical infectious diseases. 1996;23(5):1055-60.

Silveira C, Ferreira R, Muccioli C, Nussenblatt R, Belfort R. Toxoplamosis transmitted to a newborn from the mother infected 20 years earlier. American journal of ophthalmology. 2003;136(2):370-1.

Cotter TG. Apoptosis and cancer: the genesis of a research field. Nature Reviews Cancer. 2009;9(7):501-7.

Yenari MA, Liu J, Zheng Z, Vexler ZS, Lee JE, Giffard RG. Antiapoptotic and Anti‐inflammatory Mechanisms of Heat‐Shock Protein Protection. Annals of the New York Academy of Sciences. 2005;1053(1):74-83.

Hippe D, Weber A, Zhou L, Chang DC, Häcker G, Lüder CG. Toxoplasma gondii infection confers resistance against BimS-induced apoptosis by preventing the activation and mitochondrial targeting of pro-apoptotic Bax. Journal of cell science. 2009;122(19):3511-21.

Carmen JC, Hardi L, Sinai AP. Toxoplasma gondii inhibits ultraviolet light‐induced apoptosis through multiple interactions with the mitochondrion‐dependent programmed cell death pathway. Cellular microbiology. 2006;8(2):301-15.

Hwang I-Y, Quan JH, Ahn M-H, Ahmed HAH, Cha G-H, Shin D-W, et al. Toxoplasma gondii infection inhibits the mitochondrial apoptosis through induction of Bcl-2 and HSP70. Parasitology research. 2010;107(6):1313-21.

Sedletska Y, Giraud-Panis M-J, Malinge J-M. Cisplatin is a DNA-damaging antitumour compound triggering multifactorial biochemical responses in cancer cells: importance of apoptotic pathways. Current Medicinal Chemistry-Anti-Cancer Agents. 2005;5(3):251-65.

Liu Y, Xing H, Han X, Shi X, Liang F, Cheng G, et al. Apoptosis of HeLa cells induced by cisplatin and its mechanism. Journal of Huazhong University of Science and Technology [Medical Sciences]. 2008;28:197-9.

Grammatopoulos TN, Morris K, Bachar C, Moore S, Andres R, Weyhenmeyer JA. Angiotensin II attenuates chemical hypoxia-induced caspase-3 activation in primary cortical neuronal cultures. Brain research bulletin. 2004;62(4):297-303.

Tanida S, Mizoshita T, Ozeki K, Tsukamoto H, Kamiya T, Kataoka H, et al. Mechanisms of cisplatin-induced apoptosis and of cisplatin sensitivity: potential of BIN1 to act as a potent predictor of cisplatin sensitivity in gastric cancer treatment. International journal of surgical oncology. 2012;2012.

Liu Y, Xing H, Han X, Shi X, Liang F, Chen G, et al. The mechanism of cisplatin-induced apoptosis in HeLa cells. Chinese Journal of Clinical Oncology. 2005;2(6):866-9.

Wahab A, Ibrahim S, Abdul AB, Alzubairi AS, Mohamed Elhassan M, Mohan S. In vitro ultramorphological assessment of apoptosis induced by zerumbone on (HeLa). BioMed Research International. 2009;2009.

Crescitelli R, Lässer C, Szabo TG, Kittel A, Eldh M, Dianzani I, et al. Distinct RNA profiles in subpopulations of extracellular vesicles: apoptotic bodies, microvesicles and exosomes. Journal of extracellular vesicles. 2013;2.

Jones JL, Kruszon-Moran D, Wilson M, McQuillan G, Navin T, McAuley JB. Toxoplasma gondii infection in the United States: seroprevalence and risk factors. American journal of epidemiology. 2001;154(4):357-65.

Dobrowolski JM, Sibley LD. Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite. Cell. 1996;84(6):933-9.

Laliberte J, Carruthers VB. Host cell manipulation by the human pathogen Toxoplasma gondii. Cellular and molecular life sciences. 2008;65(12):1900-15.

Larasati YA, Putri DDP, Utomo RY, Hermawan A, Meiyanto E. Combination of Cisplatin and Cinnamon Essential Oil Inhibits HeLa Cells Proliferation through Cell Cycle Arrest. 2014.

Barcinski M, DosReis G. Apoptosis in parasites and parasite-induced apoptosis in the host immune system: a new approach to parasitic diseases. Brazilian Journal of Medical and Biological Research. 1999;32(4):395-401.

Kim J-Y, Ahn M-H, Jun H-S, Jung J-W, Ryu J-S, Min D-Y. Toxoplasma gondii inhibits apoptosis in infected cells by caspase inactivation and NF-κB activation. Yonsei medical journal. 2006;47(6):862-9.

Contreras-Ochoa CO, Lagunas-Martínez A, Belkind-Gerson J, Díaz-Chávez J, Correa D. Toxoplasma gondii invasion and replication within neonate mouse astrocytes and changes in apoptosis related molecules. Experimental parasitology. 2013;134(2):256-65.

Koprinarova M, Markovska P, Iliev I, Anachkova B, Russev G. Sodium butyrate enhances the cytotoxic effect of cisplatin by abrogating the cisplatin imposed cell cycle arrest. BMC molecular biology. 2010;11(1):49.

Elmore S. Apoptosis: a review of programmed cell death. Toxicologic pathology. 2007;35(4):495-516.

Shamseddin J, Akhlaghi L, Razmjou E, Shojaee S, Monavari SHR, Tajik N, et al. Conjugated Linoleic Acid Stimulates Apoptosis in RH and Tehran Strains of Toxoplasma gondii, in Vitro. Iranian journal of parasitology. 2015;10(2):238.

Kurts C, Robinson BW, Knolle PA. Cross-priming in health and disease. Nature Reviews Immunology. 2010;10(6):403-14.

Henry F, Boisteau O, Bretaudeau L, Lieubeau B, Meflah K, Grégoire M. Antigen-presenting cells that phagocytose apoptotic tumor-derived cells are potent tumor vaccines. Cancer research. 1999;59(14):3329-32.

Hsieh S-H, Lin J-S, Huang J-H, Wu S-Y, Chu C-L, Kung JT, et al. Immunization with apoptotic phagocytes containing Histoplasma capsulatum activates functional CD8+ T cells to protect against histoplasmosis. Infection and immunity. 2011;79(11):4493-502.

Files
IssueVol 13 No 3 (2018) QRcode
SectionOriginal Article(s)
Keywords
Apoptosis Apoptotic blebs Toxoplasma gondii Toxoplasmosis Cisplatin NaN3

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
CHERAGHIPOUR K, SHARIATI L, KHANAHMAD H, GANJALIKHANI-HAKEMI M, MORIDNIA A, MIRIAN M, PESTEHCHIAN N. Induction of Apoptosis in Toxoplasma gondii Infected Hela Cells by Cisplatin and Sodium Azide and Isolation of Apoptotic Bodies and Potential Use for Vaccination against Toxoplasma gondii. Iran J Parasitol. 2018;13(3):406-415.