Virologica Sinica

, Volume 32, Issue 5, pp 404–414 | Cite as

Attenuated phenotypes and analysis of a herpes simplex virus 1 strain with partial deletion of the UL7, UL41 and LAT genes

  • Xingli Xu
  • Yingqiu Guo
  • Shengtao Fan
  • Pingfang Cui
  • Min Feng
  • Lichun Wang
  • Ying Zhang
  • Yun Liao
  • Xiaolong Zhang
  • Qihan LiEmail author
Research Article


We previously constructed a herpes simplex virus 1 (HSV-1) UL7 mutant virus (M1) and showed that a partial deletion mutation of the UL7 gene led to a lower proliferative rate and an attenuated phenotype. Using the M1 mutant, we further modified the UL41 gene, which encodes another tegument protein, and the latency-associated transcript (LAT) gene. Observations of the resulting mutants with modified UL7 and UL41 (M2) or UL7, UL41 and LAT (M3) genes indicated attenuated phenotypes, with lower proliferative ratios in various cells, non-lethal infections in mice and lower viral loads in nervous tissues compared with the wild-type strain. Furthermore, no LAT stable intron could be detected in the trigeminal ganglion of M3-infected animals. The results obtained with the three HSV-1 mutants indicate that the M3 mutant is an attenuated strain with low pathogenicity during both acute and latent infections. Together, the results support the use of the M3 mutant as a candidate for the development of an HSV-1 vaccine.


herpes simplex virus 1 (HSV-1) UL7 UL41 LAT mutant 



This work was supported by the National Basic Research Program (2012CB518901), Chinese academy of medical sciences (CAMS) Initiative for Innovative Medicine (2016-I2M-1-019), the National Natural Science Foundation of China (31300143, 31100127), and the Fundamental Research Funds for the Central Universities (2016ZX310047, 2016ZX350072). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author Contributions

QHL designed the experiments; XLX, STF, PFC, LCW and XLZ performed the experiments; XLX, YZ and YL performed the analyses; XLX and MF wrote the manuscript; YQG completed the majority of the manuscript revisions and refined the English used in the manuscript, and all of the authors contributed to, read, and approved the final version of the manuscript.

Compliance with Ethics Guidelines

The authors declare that they have no conflict of interest. The experimental protocols were reviewed and approved by the Yunnan Provincial Experimental Animal Management Association (approval number: SCXK [Dian] 2011–0005) and the Experimental Animal Ethics Committee of the Institute of Medical Biology, Chinese Academy of Medical Sciences. The animal experiments were designed based on the principles expressed in the “Guide for the Care and Use of Laboratory Animals” and “Guidance for Experimental Animal Welfare and Ethical Treatment.” All institutional and national guidelines for the care and use of laboratory animals were followed.

Supplementary material

12250_2017_3947_MOESM1_ESM.pdf (2.3 mb)
Attenuated phenotypes and analysis of a herpes simplex virus 1 strain with partial deletion of the UL7, UL41 and LAT genes


  1. Aranda AM, Epstein AL. 2015. Herpes simplex virus type 1 latency and reactivation: an update. Med Sci (Paris), 31: 506–514.CrossRefGoogle Scholar
  2. Augustinova H, Hoeller D, Yao F. 2004. The dominant-negative herpes simplex virus type 1 (HSV-1) recombinant CJ83193 can serve as an effective vaccine against wild-type HSV-1 infection in mice. J Virol, 78: 5756–5765.CrossRefGoogle Scholar
  3. Awasthi S, Lubinski JM, Eisenberg RJ, Cohen GH, Friedman HM. 2008. An HSV-1 gD mutant virus as an entry-impaired live virus vaccine. Vaccine, 26: 1195–1203.CrossRefGoogle Scholar
  4. BenMohamed L, Osorio N, Srivastava R, Khan AA, Simpson JL, Wechsler SL. 2015. Decreased reactivation of a herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) mutant using the in vivo mouse UV-B model of induced reactivation. J Neurovirol, 21: 508–517.CrossRefGoogle Scholar
  5. Brehm M, Samaniego LA, Bonneau RH, DeLuca NA, Tevethia SS. 1999. Immunogenicity of herpes simplex virus type 1 mutants containing deletions in one or more alpha-genes: ICP4, ICP27, ICP22, and ICP0. Virology, 256: 258–269.CrossRefGoogle Scholar
  6. Cotter CR, Nguyen ML, Yount JS, López CB, Blaho JA, Moran TM. 2010. The virion host shut-off (vhs) protein blocks a TLRindependent pathway of herpes simplex virus type 1 recognition in human and mouse dendritic cells. PLoS One, 5: e8684.CrossRefGoogle Scholar
  7. David AT, Saied A, Charles A, Subramanian R, Chouljenko VN, Kousoulas KG. 2012. A herpes simplex virus 1 (McKrae) mutant lacking the glycoprotein K gene is unable to infect via neuronal axons and egress from neuronal cell bodies. MBio, 3(e00144-e00112): e00144–00112.PubMedPubMedCentralGoogle Scholar
  8. Dumitrascu OM, Mott KR, Ghiasi H. 2014. A comparative study of experimental mouse models of central nervous system demyelination. Gene Ther, 21: 599–608.CrossRefGoogle Scholar
  9. Farrell MJ, Dobson AT, Feldman LT. 1991. Herpes simplex virus latency-associated transcript is a stable intron. Proc Natl Acad Sci USA, 88: 790–794.CrossRefGoogle Scholar
  10. Fatahzadeh M, Schwartz RA. 2007. Human herpes simplex virus infections: epidemiology, pathogenesis, symptomatology, diagnosis, and management. J Am Acad Dermatol, 57: 737–63; quiz 764.CrossRefGoogle Scholar
  11. Garland SM, Steben M. 2014. Genital herpes. Best Pract Res Clin Obstet Gynaecol, 28: 1098–1110.CrossRefGoogle Scholar
  12. Herrera FJ, Triezenberg SJ. 2004. VP16-dependent association of chromatin-modifying coactivators and underrepresentation of histones at immediate-early gene promoters during herpes simplex virus infection. J Virol, 78: 9689–9696.CrossRefGoogle Scholar
  13. Johnston C, Gottlieb SL, Wald A. 2016. Status of vaccine research and development of vaccines for herpes simplex virus. Vaccine, 34: 2948–2952.CrossRefGoogle Scholar
  14. Kelly BJ, Fraefel C, Cunningham AL, Diefenbach RJ. 2009. Functional roles of the tegument proteins of herpes simplex virus type 1. Virus Res, 145: 173–186.CrossRefGoogle Scholar
  15. Koelle DM, Corey L. 2003. Recent progress in herpes simplex virus immunobiology and vaccine research. Clin Microbiol Rev, 16: 96–113.CrossRefGoogle Scholar
  16. Kukhanova MK, Korovina AN, Kochetkov SN. 2014. Human herpes simplex virus: life cycle and development of inhibitors. Biochemistry (Mosc), 79: 1635–1652.CrossRefGoogle Scholar
  17. Lee K, Kolb AW, Larsen I, Craven M, Brandt CR. 2016. Mapping murine corneal neovascularization and weight loss virulence determinants in the herpes simplex virus 1 genome and the detection of an epistatic interaction between the UL and IRS/US regions. J Virol, 90: 8115–8131.CrossRefGoogle Scholar
  18. Leib DA, Bogard CL, Kosz-Vnenchak M, Hicks KA, Coen DM, Knipe DM, Schaffer PA. 1989. A deletion mutant of the latency-associated transcript of herpes simplex virus type 1 reactivates from thelatent state with reduced frequency. J Virol, 63: 2893–2900.PubMedPubMedCentralGoogle Scholar
  19. Looker KJ, Magaret AS, May MT, Turner KM, Vickerman P, Gottlieb SL, Newman LM. 2015. Global and Regional Estimates of Prevalent and Incident Herpes Simplex Virus Type 1 Infections in 2012. PLoS One, 10: e0140765.CrossRefGoogle Scholar
  20. Loret S, Lippé R. 2012. Biochemical analysis of infected cell polypeptide (ICP)0, ICP4, UL7 and UL23 incorporated into extracellular herpes simplex virus type 1 virions. J Gen Virol, 93: 624–634.CrossRefGoogle Scholar
  21. Maggioncalda J, Mehta A, Fraser NW, Block TM. 1994. Analysis of a herpes simplex virus type 1 LAT mutant with a deletion between the putative promoter and the 5′ end of the 2. 0-kilobase TranScript. J Virol, 68: 7816–7824.PubMedGoogle Scholar
  22. Morrison LA, Knipe DM. 1997. Contributions of antibody and T cell subsets to protection elicited by immunization with a replication-defective mutant of herpes simplex virus type 1. Virology, 239: 315–326.CrossRefGoogle Scholar
  23. Paludan SR, Bowie AG, Horan KA, Fitzgerald KA. 2011. Recognition of herpesviruses by the innate immune system. Nat Rev Immunol, 11: 143–154.CrossRefGoogle Scholar
  24. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. 2013. Genome engineering using the CRISPR-Cas9 system. Nat Protoc, 8: 2281–2308.CrossRefGoogle Scholar
  25. Read GS, Karr BM, Knight K. 1993. Isolation of a herpes simplex virus type 1 mutant with a deletion in the virion host shutoff gene and identification of multiple forms of the vhs (UL41) polypeptide. J Virol, 67: 7149–7160.PubMedPubMedCentralGoogle Scholar
  26. Saffran HA, Read GS, Smiley JR. 2010. Evidence for translational regulation by the herpes simplex virus virion host shutoff protein. J Virol, 84: 6041–6049.CrossRefGoogle Scholar
  27. Samady L, Costigliola E, MacCormac L, McGrath Y, Cleverley S, Lilley CE, Smith J, Latchman DS, Chain B, Coffin RS. 2003. Deletion of the virion host shutoff protein (vhs) from herpes simplex virus (HSV) relieves the viral block to dendritic cell activation: potential of vhs-HSV vectors for dendritic cellmediated immunotherapy. J Virol, 77: 3768–3776.CrossRefGoogle Scholar
  28. Samoto K, Perng GC, Ehtesham M, Liu Y, Wechsler SL, Nesburn AB, Black KL, Yu JS. 2001. A herpes simplex virus type 1 mutant deleted for gamma34.5 and LAT kills glioma cells in vitro and is inhibited for in vivo reactivation. Cancer Gene Ther, 8: 269–277.CrossRefGoogle Scholar
  29. Sawtell NM, Triezenberg SJ, Thompson RL. 2011. VP16 serine 375 is a critical determinant of herpes simplex virus exit from latency in vivo. J Neurovirol, 17: 546–551.CrossRefGoogle Scholar
  30. Stanfield B, Kousoulas KG. 2015. Herpes Simplex Vaccines: Prospects of Live-attenuated HSV Vaccines to Combat Genital and Ocular infections. Curr Clin Microbiol Rep, 2: 125–136.CrossRefGoogle Scholar
  31. Strain AK, Rice SA. 2011. Phenotypic suppression of a herpes simplex virus 1 ICP27 mutation by enhanced transcription of the mutant gene. J Virol, 85: 5685–5690.CrossRefGoogle Scholar
  32. Tanaka M, Sata T, Kawaguchi Y. 2008. The product of the herpes simplex virus 1 UL7 gene interacts with a mitochondrial protein, adenine nucleotide translocator 2. Virol J, 5: 125.CrossRefGoogle Scholar
  33. Thompson RL, Sawtell NM. 1997. The herpes simplex virus type 1 latency-associated transcript gene regulates the establishment of latency. J Virol, 71: 5432–5440.PubMedPubMedCentralGoogle Scholar
  34. Wagner EK, Flanagan WM, Devi-Rao G, Zhang YF, Hill JM, Anderson KP, Stevens JG. 1988. The herpes simplex virus latencyassociated transcript is spliced during the latent phase of infection. J Virol, 62: 4577–4585.PubMedPubMedCentralGoogle Scholar
  35. Xu F, Sternberg MR, Kottiri BJ, McQuillan GM, Lee FK, Nahmias AJ, Berman SM, Markowitz LE. 2006. Trends in herpes simplex virus type 1 and type 2 seroprevalence in the United States. JAMA, 296: 964–973.CrossRefGoogle Scholar
  36. Xu X, Che Y, Li Q. 2016a. HSV-1 tegument protein and the development of its genome editing technology. Virol J, 13: 108.CrossRefGoogle Scholar
  37. Xu X, Fan S, Zhou J, Zhang Y, Che Y, Cai H, Wang L, Guo L, Liu L, Li Q. 2016b. The mutated tegument protein UL7 attenuates the virulence of herpes simplex virus 1 by reducing the modulation of alpha-4 gene transcription. Virol J, 13: 152.CrossRefGoogle Scholar
  38. Yu X, Liu L, Wu L, Wang L, Dong C, Li W, Li Q. 2010. Herpes simplex virus type 1 tegument protein VP22 is capable of modulating the transcription of viral TK and gC genes via interaction with viral ICP0. Biochimie, 92: 1024–1030.CrossRefGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS and Springer Nature Singapore Pte Ltd. 2017

Authors and Affiliations

  1. 1.Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical CollegeYunnan Key Laboratory of Vaccine Research and Development of Severe Infectious DiseaseKunmingChina

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