Volume 5, Issue 2 (Summer-Fall 2022)
Mod Med Lab J 2022, 5(2): 61-69 |
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Parsa M, Zargan J, Honari H, Hajinourmohammadi A, Mousavi S M, Keshavarz Alikhani H. In-vitro Evaluation of the Antibacterial and Cytotoxicity Activity of the PAD4 Antigen of Bacillus anthracis as a vaccine candidate. Mod Med Lab J 2022; 5 (2) :61-69
URL: http://modernmedlab.com/article-1-119-en.html
URL: http://modernmedlab.com/article-1-119-en.html
Mohsen Parsa 
, Jamil Zargan , Hossein Honari , Ashkan Hajinourmohammadi , Seyyed Mohsen Mousavi , Hani Keshavarz Alikhani
, Jamil Zargan , Hossein Honari , Ashkan Hajinourmohammadi , Seyyed Mohsen Mousavi , Hani Keshavarz Alikhani
Abstract: (1330 Views)
Introduction: Infectious diseases are one of the main causes of death worldwide. This has driven scientists to invest in extraction and identification of antimicrobial agents from natural toxins and presentation of novel antibiotics and vaccines. The aim of the current study is to investigate the antibacterial and cytotoxicity effects of the protective antigen domain 4 (PAD4) from Bacillus anthracis as a strong immunogen and vaccine candidate for B. anthracis.
Matherial and Methods: In this study, the antibacterial effect of the antigen was evaluated in concentrations of 0.28-4.5μg/ml using MTT reduction and MIC assays and the anticancer effect of the recombinant PAD4 on MCF-7 cell line was examined in concentrations of 0.5-2μg/ml via MTT, neutral red uptake, and comet assays. NO, GSH and catalase determination assays following the treatment with PAD4 was also evaluated.
Results: According to the antibacterial results, PAD4 did not show any antibacterial effect against S. aureus, but very little inhibition on E. coli cells' growth was recorded. The results of MTT and neutral red assays showed that this antigen has a significant inhibiting effect on cancer cell growth. Comet assay results showed that PAD4 can cause death of breast cancer cells by apoptosis induction. NO, GSH and catalase determination assays did not show any significant fluctuations following the treatment with PAD4.
Conclusion: Our results showed that this antigen does not have any antibacterial effect but it can inhibit the proliferation of breast cancer cells, making PAD4 a candidate for producing antitumor drugs.
Matherial and Methods: In this study, the antibacterial effect of the antigen was evaluated in concentrations of 0.28-4.5μg/ml using MTT reduction and MIC assays and the anticancer effect of the recombinant PAD4 on MCF-7 cell line was examined in concentrations of 0.5-2μg/ml via MTT, neutral red uptake, and comet assays. NO, GSH and catalase determination assays following the treatment with PAD4 was also evaluated.
Results: According to the antibacterial results, PAD4 did not show any antibacterial effect against S. aureus, but very little inhibition on E. coli cells' growth was recorded. The results of MTT and neutral red assays showed that this antigen has a significant inhibiting effect on cancer cell growth. Comet assay results showed that PAD4 can cause death of breast cancer cells by apoptosis induction. NO, GSH and catalase determination assays did not show any significant fluctuations following the treatment with PAD4.
Conclusion: Our results showed that this antigen does not have any antibacterial effect but it can inhibit the proliferation of breast cancer cells, making PAD4 a candidate for producing antitumor drugs.
Type of Study: Original Research Article |
Subject:
Immunology
References
1. Schmidt TR, Scott EJ, Dyer DW. Whole-genome phylogenies of the family Bacillaceae and expansion of the sigma factor gene family in the Bacillus cereus species-group. BMC genomics. 2011;12(1):1-17.
2. Read TD, Salzberg SL, Pop M, Shumway M, Umayam L, Jiang L, et al. Comparative genome sequencing for discovery of novel polymorphisms in Bacillus anthracis. Science. 2002;296(5575):2028-33.
3. Khanna H, Singh Y. War against anthrax. Molecular Medicine. 2001;7(12):795-6.
4. Varshney A, Kumar M, Nagar D, Pal V, Goel A. Development of a novel chimeric PA-LF antigen of Bacillus anthracis, its immunological characterization and evaluation as a future vaccine candidate in mouse model. Biologicals. 2019;61:38-43.
5. Okinaka R, Cloud K, Hampton O, Hoffmaster A, Hill K, Keim P, et al. Sequence, assembly and analysis of pX01 and pX02. Journal of Applied Microbiology. 1999;87(2):261-2.
6. Santelli E, Bankston LA, Leppla SH, Liddington RC. Crystal structure of a complex between anthrax toxin and its host cell receptor. Nature. 2004;430(7002):905-8.
7. Brossier F, Weber-Levy M, Mock M, Sirard J-C. Role of toxin functional domains in anthrax pathogenesis. Infection and immunity. 2000;68(4):1781-6.
8. Manish M, Verma S, Kandari D, Kulshreshtha P, Singh S, Bhatnagar R. Anthrax prevention through vaccine and post-exposure therapy. Expert Opinion on Biological Therapy. 2020;20(12):1405-25.
9. Rezaee M, Honari H, Kooshk MRA. Cloning, expression and purification of binding domains of lethal factor and protective antigen of Bacillus anthracis in Escherichia coli and evaluation of their related murine antibody. Molecular biology reports. 2014;41(4):2445-52.
10. MehrAzin H, Honari H, Saadati M, Alizadeh H, Nazarian S. Production of polyclonal antibody against domain 4 of protective antigen of Bacillus anthracis in laboratory animals. Passive Defence Sci. 2011;2:19-25.
11. Wang H, Cheng H, Wang F, Wei D, Wang X. An improved 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) reduction assay for evaluating the viability of Escherichia coli cells. Journal of microbiological methods. 2010;82(3):330-3.
12. Veiga A, Maria da Graça TT, Rossa LS, Mengarda M, Stofella NC, Oliveira LJ, et al. Colorimetric microdilution assay: Validation of a standard method for determination of MIC, IC50%, and IC90% of antimicrobial compounds. Journal of microbiological methods. 2019;162:50-61.
13. Repetto G, Del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nature protocols. 2008;3(7):1125-31.
14. Collins AR. The comet assay for DNA damage and repair. Molecular biotechnology. 2004;26(3):249-61.
15. Sun J, Zhang X, Broderick M, Fein H. Measurement of nitric oxide production in biological systems by using Griess reaction assay. Sensors. 2003;3(8):276-84.
16. Owen JB, Butterfield DA. Measurement of oxidized/reduced glutathione ratio. Protein misfolding and cellular stress in disease and aging: Springer; 2010. p. 269-77.
17. Montavon P, Kukic KR, Bortlik K. A simple method to measure effective catalase activities: optimization, validation, and application in green coffee. Analytical biochemistry. 2007;360(2):207-15.
18. Kaur M, Singh S, Bhatnagar R. Anthrax vaccines: present status and future prospects. Expert review of vaccines. 2013;12(8):955-70.
19. Flick-Smith HC, Walker NJ, Gibson P, Bullifent H, Hayward S, Miller J, et al. A recombinant carboxy-terminal domain of the protective antigen of Bacillus anthracis protects mice against anthrax infection. Infection and immunity. 2002;70(3):1653-6.
20. Singh Y, Khanna H, Chopra AP, Mehra V. A Dominant Negative Mutant of Bacillus anthracisProtective Antigen Inhibits Anthrax Toxin Action in Vivo. Journal of Biological Chemistry. 2001;276(25):22090-4.
21. Albrecht MT, Li H, Williamson ED, LeButt CS, Flick-Smith HC, Quinn CP, et al. Human monoclonal antibodies against anthrax lethal factor and protective antigen act independently to protect against Bacillus anthracis infection and enhance endogenous immunity to anthrax. Infection and immunity. 2007;75(11):5425-33.
22. Lin C-G, Kao Y-T, Liu W-T, Huang H-H, Chen K-C, Wang T-M, et al. Cytotoxic effects of anthrax lethal toxin on macrophage-like cell line J774A. 1. Current microbiology. 1996;33(4):224-7.
23. Rogers MS, Christensen KA, Birsner AE, Short SM, Wigelsworth DJ, Collier RJ, et al. Mutant anthrax toxin B moiety (protective antigen) inhibits angiogenesis and tumor growth. Cancer research. 2007;67(20):9980-5.
24. Liu S, Bugge TH, Leppla SH. Targeting of tumor cells by cell surface urokinase plasminogen activator-dependent anthrax toxin. Journal of Biological Chemistry. 2001;276(21):17976-84.
25. Cui X, Moayeri M, Li Y, Li X, Haley M, Fitz Y, et al. Lethality during continuous anthrax lethal toxin infusion is associated with circulatory shock but not inflammatory cytokine or nitric oxide release in rats. American Journal of Physiology-Regulatory, Integrative and Comparative Physiology. 2004;286(4):R699-R709.
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