Original Article

In vitro evaluation of inhibitory effect of Lactobacillus reuteri supernatant on the replication of herpes simplex virus type 1 and expression of UL54, UL52 and UL27 genes

Abstract

Background and Objectives: Human herpes virus type 1 (HSV-1) is a neurotropic pathogen that is infected more than 70% of the world population. The increasing of viral resistance to antiviral drugs and the emergence of side effects has motivated researchers to study the use of probiotics as new antiviral agents. The aim of the present study was to study for the first time the potential antiviral activity of Lactobacillus reuteri (L. reuteri) supernatant against HSV-1.
Materials and Methods: After measuring the cytotoxicity of L. reuteri supernatant by MTT assay, 1:16 dilution of it was added to HeLa cells before and after HSV-1 infection, after 1.5 hours incubation with HSV-1, and simultaneously with HSV-1 infection. After 48 hours of incubation at 37°C, the viral titer and expression levels of UL54, UL52 and UL27 genes were measured by tissue culture infectious dose 50 (TCID50) and Real-Time PCR methods, respectively.
Results: HSV-1 titer in the treatment conditions before infection, incubation with HSV-1, simultaneously with infection and after infection was reduced by 0.42, 3.42, 1.83, and 0.83 log 10 TCID50/ml, respectively. When the bacterial supernatant was first incubated with the virus and then added to the cell, or when it was added simultaneously with the virus, the expression of the UL27, UL52, and UL54 genes decreased significantly (p<0.05). When the bacterial supernatant is added to the cell before or after virus infection, the expression of UL52 and UL54 genes does not change significantly (P>0.05).
Conclusion: The study findings indicated that the supernatant of L. reuteri has a potent anti-HSV-1 effect especially if it is incubated with the virus before inoculation into the cell. Its possible antiviral mechanism is to inhibit the virus by binding to it or changing the surface structure of the virus. Metabolites of L. reuteri can be considered as a novel inhibitor of HSV-1infection.

1. Barzoki MG, Malekshahi SS, Shayestehpour M. In vitro evaluation of antiviral activity of Shouchella clausii probiotic strain and bacterial supernatant against herpes simplex virus type 1. Arch Microbiol 2022; 204: 522.
2. Shayestehpour M, Rahimi MR, Piroozmand A, Khaledi A, Fateminasab ZS. In vitro evaluation of antiviral activity effect of selenium, Bacillus clausii supernatant, and their combination on the replication of herpes simplex virus 1. Jundishapur J Microbiol 2022; 15(8): e129848.
3. Birkmann A, Bonsmann S, Kropeit D, Pfaff T, Rangaraju M, Sumner M, et al. Discovery, chemistry, and preclinical development of pritelivir, a novel treatment option for acyclovir-resistant Herpes simplex virus infections. J Med Chem 2022; 65: 13614-13628.
4. Schalkwijk HH, Snoeck R, Andrei G. Acyclovir resistance in herpes simplex viruses: Prevalence and therapeutic alternatives. Biochem Pharmacol 2022; 206: 115322.
5. Valladares HC, Rudd N, Leslie K. P76 Management of aciclovir-resistant mucocutaneous herpes simplex virus infection: a systematic review. Br J Dermatol 2023; 188(Supplement_4): ljad113.104.
6. Abd Wahab NZ, Ganasen T, Rahman NIA, Ibrahim N. In vitro virucidal activity of Kyllinga nemoralis aqueous extract against herpes simplex virus. J Appl Biol Biotechnol 2023; 11: 208-213.
7. Elebeedy D, Ghanem A, Aly SH, Ali MA, Faraag AH, El-Ashrey MK, et al. Synergistic antiviral activity of Lactobacillus acidophilus and Glycyrrhiza glabra against Herpes Simplex-1 Virus (HSV-1) and Vesicular Stomatitis Virus (VSV): experimental and In Silico insights. BMC Microbiol 2023; 23: 173.
8. Lasanen T, Frejborg F, Lund LM, Nyman MC, Orpana J, Habib H, et al. Single therapeutic dose of an antiviral UL29 siRNA swarm diminishes symptoms and viral load of mice infected intranasally with HSV‐1. Smart Med 2023; 2(2): e20230009.
9. Ng QX, Lim YL, Yaow CYL, Ng WK, Thumboo J, Liew TM. Effect of probiotic supplementation on gut microbiota in patients with major depressive disorders: A systematic review. Nutrients 2023; 15: 1351.
10. Kullar R, Goldstein EJ, Johnson S, McFarland LV. Lactobacillus bacteremia and probiotics: A review. Microorganisms 2023; 11: 896.
11. Mu Q, Tavella VJ, Luo XM. Role of Lactobacillus reuteri in human health and diseases. Front Microbiol 2018; 9: 757.
12. Kassaa IA, Hober D, Hamze M, Caloone D, Dewilde A, Chihib N-E, et al. Vaginal Lactobacillus gasseri CMUL57 can inhibit herpes simplex type 2 but not Coxsackievirus B4E2. Arch Microbiol 2015; 197: 657-664.
13. Khani S, Motamedifar M, Golmoghaddam H, Hosseini HM, Hashemizadeh Z. In vitro study of the effect of a probiotic bacterium Lactobacillus rhamnosus against herpes simplex virus type 1. Braz J Infect Dis 2012; 16: 129-135.
14. Mousavi E, Makvandi M, Teimoori A, Ataei A, Ghafari S, Samarbaf-Zadeh A. Antiviral effects of Lactobacillus crispatus against HSV-2 in mammalian cell lines. J Chin Med Assoc 2018; 81: 262-267.
15. Wang Y-Y, Lyu Y-N, Xin H-Y, Cheng J-T, Liu X-Q, Wang X-W, et al. Identification of putative UL54 (ICP27) transcription regulatory sequences binding to Oct-1, v-Myb, Pax-6 and hairy in herpes simplex viruses. J Cancer 2019; 10: 430-440.
16. Bermek O, Williams RS. The three-component helicase/primase complex of herpes simplex virus-1. Open Biol 2021; 11: 210011.
17. Li P, Gandhi D, Mutas M, Ran Y-F, Carr M, Rampini S, et al. Direct identification of the herpes simplex virus UL27 gene through single particle manipulation and optical detection using a micromagnetic array. Nanoscale 2020; 12: 3482-3490.
18. Ramakrishnan MA. Determination of 50% endpoint titer using a simple formula. World J Virol 2016; 5: 85-86.
19. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402-408.
20. Ranjbar R, Goudarzi MM, Jounaidi N. Lactobacillus acidophilus and assessment for its antiviral effect against herpes simplex virus type I. Biosci Biotechnol Res Asia 2015; 12: 1351-1356.
21. Al Kassaa I, Hober D, Hamze M, Chihib NE, Drider D. Antiviral potential of lactic acid bacteria and their bacteriocins. Probiotics Antimicrob Proteins 2014; 6: 177-185.
22. Ang LY, Too HK, Tan EL, Chow TK, Shek LP, Tham EH, et al. Antiviral activity of Lactobacillus reuteri protectis against Coxsackievirus A and enterovirus 71 infection in human skeletal muscle and colon cell lines. Virol J 2016; 13: 111.
23. Botić T, Klingberg TD, Weingartl H, Cencič A. A novel eukaryotic cell culture model to study antiviral activity of potential probiotic bacteria. Int J Food Microbiol 2007; 115: 227-234.
24. Vilhelmova-Ilieva N, Atanasov G, Simeonova L, Dobreva L, Mancheva K, Trepechova M, et al. Anti-herpes virus activity of lactobacillus' postbiotics. Biomedicine (Taipei) 2022; 12: 21-29.
25. Xiao J, Cai M, Wang Y, Ding P. Antiviral activities of officinaloside C against Herpes simplex Virus-1. Molecules 2022; 27: 3365.
26. Fakhredini K, Soleimanjahi H, Bamdad T. Prevalence of herpes simplex viruses types 1 and 2 infections among suspected children of encephalitis in Kermanshah, Iran. Iran J Microbiol 2023; 15: 149-154.
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IssueVol 16 No 1 (2024) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v16i1.14877
Keywords
Antiviral; Cell culture; Lactobacillus reuteri; Herpes simplex virus type 1

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How to Cite
1.
Ebneali F, Shayestehpour M, Piroozmand A, Sedaghat H, Yazdani S, Fateminasab Z. In vitro evaluation of inhibitory effect of Lactobacillus reuteri supernatant on the replication of herpes simplex virus type 1 and expression of UL54, UL52 and UL27 genes. Iran J Microbiol. 2024;16(1):90-96.