Inactivation of the UL37 Deamidase Enhances Virus Replication and Spread of the HSV-1(VC2) Oncolytic Vaccine Strain and Secretion of GM-CSF

• Published in: Viruses 2023-01, Vol.15 (2), p.367
• Main Authors: Clark, Carolyn M, Jambunathan, Nithya, Collantes, Therese M A, Kousoulas, Konstantin G
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Breast cancer; Cancer; Cell culture; Cerebrospinal fluid; Cytokines; Enzyme inhibitors; Epithelial cells; Fibroblasts; Genes; Genetic aspects; glycoprotein K; GM-CSF; Granulocyte-Macrophage Colony-Stimulating Factor; Guinea Pigs; Herpes simplex virus; Herpes viruses; herpesvirus; Herpesvirus 1, Human - genetics; Infections; Laboratory animals; Latency; Male; Melanoma; Melanoma - therapy; Membrane proteins; Membranes; Mice; Oncolysis; Pathogens; Physiological aspects; Plaques; Prostate cancer; Proteins; Replication; Tegument; Tumor cell lines; UL20; UL37; Vaccines; Vaccines, Attenuated; VC2; Virions; Virulence; Virus Replication; Virus research; Viruses
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v15020367

The HSV-1 (VC2) live-attenuated vaccine strain was engineered with specific deletions in the amino termini of glycoprotein K (gK) and membrane protein UL20, rendering the virus unable to enter neurons and establish latency. VC2 replicates efficiently in epithelial cell culture but produces lower viral titers and smaller viral plaques than its parental HSV-1 (F) wild-type virus. VC2 is an effective live-attenuated vaccine against HSV-1 and HSV-2 infections in mice and guinea pigs and an anti-tumor immunotherapeutic and oncolytic virus against melanoma and breast cancer in mouse models. Previously, we reported that the gK/UL20 complex interacts with the UL37 tegument protein, and this interaction is essential for virion intracellular envelopment and egress. To investigate the potential role of the UL37 deamidase functions, the recombinant virus FC819S and VC2C819S were constructed with a C819S substitution to inactivate the UL37 predicted deamidase active site on an HSV-1(F) and HSV-1(VC2) genetic background, respectively. FC819S replicated to similar levels with HSV-1(F) and produced similar size viral plaques. In contrast, VC2C819S replication was enhanced, and viral plaques increased in size, approaching those of the wild-type HSV-1(F) virus. FC819S infection of cell cultures caused enhanced GM-CSF secretion in comparison to HSV-1(F) across several cell lines, including HEp2 cells and cancer cell lines, DU145 (prostate) and Panc 04.03 (pancreas), and primary mouse peritoneal cells. VC2 infection of these cell lines caused GM-CSF secretion at similar levels to FC819S infection. However, the VC2C819S virus did not exhibit any further enhancement of GM-CSF secretion compared to the VC2 virus. These results suggest that the UL37 deamidation functions in conjunction with the gK/UL20 complex to facilitate virus replication and GM-CSF secretion.