Abstract
Several antimicrobial agents are commonly included in contact lens disinfectant solutions including chlorhexidine diacetate (CHX), polyhexamethylene biguanide (PHMB) or myristamidopropyl dimethylamine (MAPD); however, their mode of action, i.e. necrosis versus apoptosis is incompletely understood. Here, we determined whether a mechanism of cell death resembling that of apoptosis was present in Acanthamoeba castellanii of the T4 genotype (NEFF) following exposure to the aforementioned antimicrobials using the anticoagulant annexin V that undergoes rapid high affinity binding to phosphatidylserine in the presence of calcium, making it a sensitive probe for phosphatidylserine exposure. The results revealed that under the conditions employed in this study, an apoptotic pathway of cell death in this organism at the tested conditions does not occur. Our findings suggest that necrosis is the likely mode of action; however, future mechanistic studies should be accomplished in additional experimental conditions to further comprehend the molecular mechanisms of cell death in Acanthamoeba.
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References
Akbar N, Hussain K, Khalid M, Siddiqui R, Shah MR, Khan NA (2023) Azole and 5-nitroimidazole based nanoformulations are potential antiamoebic drug candidates against brain-eating amoebae. J Appl Microbiol 134(4):lxad072
Ameisen JC (2002) On the origin, evolution, and nature of programmed cell death: a timeline of four billion years. Cell Death Differ 9:367–393
Arnoult D, Akarid K, Grodet A, Petit PX, Estaquier J, Ameisen JC (2002) On the evolution of programmed cell death: apoptosis of the unicellular eukaryote Leishmania major involves cysteine proteinase activation and mitochondrion permeabilisation. Cell Death Differ 9:65–81
Baig AM, Lalani S, Khan NA (2017) Apoptosis in Acanthamoeba castellanii belonging to thr T4 genotype. J Basic Microbiol 57(7):574–579
Bar PR (1996) Apoptosis – the cells’ silent exit. Life Sci 59:369–378
Biagini GA, Finlay BJ, Lloyd D (1997) Evolution of the hydrogenosome. FEMS Microbiol Lett 155:133–140
Chose O, Noel C, Gerbod D, Brenner C, Viscogliosi E, Roseto A (2002) A form of cell death with some features resembling apoptosis in the amitochondrial unicellular organism Trichomonas vaginalis. Exp Cell Res 276:32–39
Dyall SD, Johnson PJ (2000) Origins of hydrogenosomes and mitochondria: evolution and organelle biogenesis. Curr Opin Microbiol 3:404–411
Elsheikha HM, Siddiqui R, Khan NA (2020) Drug discovery against Acanthamoeba infections: present knowledge and unmet needs. Pathogens 9(5):405
Frohlich K-U, Madeo F (2001) Apoptosis in yeast: a new model for aging research. Exp Gerontol 37:27–31
Fuchs S, Hollins A, Laue M, Schaefer U, Roemer K, Gumbleton M, Lehr CM (2003) Differentiation of human alveolar epithelial cells in primary culture: morphological characterization and synthesis of caveolin-1 and surfactant protein-C. Cell Tissue Res 311:31–45
Hengartner MO (2000) The biochemistry of apoptosis. Nature 407:770–776
Kam PCA, Ferch NI (2000) Apoptosis: mechanisms and clinical implications. Anaesthesia 55:1081–1093
Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, Oers MH (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415–1420
Lee HA, Park SM, Chu KB, Quan FS, Kurz T, Pflieger M, Moon EK (2020) Application of histone deacetylase inhibitors MPK472 and KSK64 as a potential treatment for Acanthamoeba keratitis. Antimicrob Agents Chemother 64(12):e10506–e10520
Madeo F, Frohlich E, Frohlich K-U (1997) A yeast mutant showing diagnostic markers of early and late apoptosis. J Cell Biol 139:729–734
Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. As agents of disease in humans. Clin Microbiol Rev 16:273–307
Moon EK, Choi HS, Kong HH, Quan FS (2018) Polyhexamethylene biguanide and chloroquine induce programmed cell death in Acanthamoeba castellanii. Exp Parasitol 191:31–35
Quan FS, Lee HA, Kong HH, Moon EK (2020) Histone deacetylase Inhibitors enhance the amoebicidal effect of low concentrations of polyhexamethylene biguanide by inducing apoptosis. Cornea 39(2):245–249
Rodriguez-Exposito RL, Sifouij I, Reyos-Batelle M, Maciver SK, Pinero JE, Lorenzo-Morales J (2022) Statins induce actin cytoskeleton disassembly and an apoptosis-like process in Acanthamoeba spp. Antibiotics (Basel) 11(2):280
Siddiqui R, Aqeel Y, Khan NA (2016) The development of drugs against Acanthamoeba infections. Antimicrob Agents Chemother 60(11):6441–6450
Siddiqui R, Makhlouf Z, Akbar N, Khamis M, Ibrahim T, Khan AS, Khan NA (2023) Antiamoebic properties of methyltrioctylammonium chloride based deep eutectic solvents. Cont Lens Anterior Eye 46(2):101758
Sifaoui I, Lopez-Arencibia A, Martín-Navarro CM, Reyes-Batlle M, Wagner C, Chiboub O, Mejri M, Valladares B, Abderrabba M, Pinero JE, Lorenzo-Morales J (2017) Programmed cell death in Acanthamoeba castellanii Neff induced by several molecules present in olive leaf extracts. PloS One 12(8):e0183795
Somersan S, Bhardwaj N (2001) Tethering and tickling: a new role for the phosphatidylserine receptor. J Cell Biol 155:501–504
Turner NA, Russell AD, Furr JR, Lloyd D (1999) Acanthamoeba spp., antimicrobial agents and contact lenses. Sci Prog 82:1–8
Turner NA, Russell AD, Furr JR, Lloyd D (2000) Emergence of resistance to biocides during differentiation of Acanthamoeba castellanii. J Antimicrob Chemother 46:27–34
van Gurp M, Festjens N, van Loo G, Saelens X, Vandenabeele P (2003) Mitochondrial intermembrane proteins in cell death. Biochem Biophys Res Commun 304:487–497
Vaux DL, Strasser A (1996) The molecular biology of apoptosis. Proc Natl Acad Sci USA 93:2239–2244
Vermes I, Haanen C, Reutelingsperger C (2000) Flow cytometry of apoptotic cell death. J Immunol Methods 243:167–190
Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C (1995) A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods 184:39–51
Wu D, Qiao K, Feng M, Fu Y, Cai J, Deng ZY, Tacibana H, Cheng X (2018) Apoptosis of Acanthamoeba castellanii trophozoitees induced by oleic acid. J Eukaryot Microbiol 65(2)):191–199
Zangger H, Mottram JC, Fasel N (2002) Cell death in Leishmania induced by stress and differentiation: programmed cell death or necrosis? Cell Death Differ 9:1126–1139
Acknowledgements
The authors wish to thank Alcon Research Ltd., Fort Worth, Texas 76134-2099, USA, for providing the MAPD and also Dr. Andrew Hollins, Welsh School of Pharmacy, for his help with the flow cytometry.
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Ruqaiyyah Siddiqui and Naveed Ahmed Khan are supported by the Air Force Office of Scientific Research (AFOSR), USA.
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LT and DL conceptualised the study amid discussions with RS and NAK. LT, RS and BSA carried out all bioassays under the supervision of NAK and DL. LT and DL analysed the data and prepared the first draft of the manuscript. RS, BSA and NAK revised and corrected the manuscript. All authors approved the final manuscript.
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Thomas, L., Khan, N.A., Siddiqui, R. et al. Cell death of Acanthamoeba castellanii following exposure to antimicrobial agents commonly included in contact lens disinfecting solutions. Parasitol Res 123, 16 (2024). https://doi.org/10.1007/s00436-023-08061-5
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DOI: https://doi.org/10.1007/s00436-023-08061-5