Characterization of strain HSZP of herpes simplex virus type 1 (HSV1)

The generic background of HSZP virus, an HSV1 strain with extensive passage history, was analyzed by parallel comparative sequencing of four relevant genes (UL27/gB, UL41/vhs, UL44/gC and UL53/gK) of HSZP and additional three selected viruses [strains ANGpath, strains KOS(a) and KOS(b) and the prototype strain 17]. Mutation at position 858 (His for Arg) in gB of HSZP was found to be responsible for giant cell formation (syn(3)gB mutation) similarly as the 855 mutation (Val for Ala) in the gB of ANGpath. No syn(1)gK mutations were detected in the UL53 gene either of HSZP or of ANGpath viruses. The reduced virulence of HSZP for adult mice after peripheral inoculation, similarly as that of KOS virus, seems to be related (at least in part) to numerous mutations in the gB ectodomain. Of these, two mutations located in the antigenic domain IV were the same in gB(HSZP) as well as in gB(KOS) (at amino acids 59 and 79), at least two (amino acids 313 and 553) were specific for gB(KOS), while one mutation (Ser for Ala at position 108) was specific for gB(HSZP). Th, abolished shutoff function of the HSZP virus was related to at least four out of six specific mutations seen in the vhs polypeptide (vhr(HSZP)) encoded by the UL41 gene, of which three (amino acids 374, 386, 392) were clustered in the semiconservative box A of vhr(HSZP) (the truncation of which abrogates the inhibition provided by this protein) and one mutation (at amino acid 18) was situated in the highly conservative locus I of vhs(HSZP). In addition, the two vhs(KOS) specific mutations (amino acids 19 and 317) not found in vhs(HSZP), enhanced the early host shutoff function of the vhs(KOS) protein. Finally, gC(HSZP) had two specific mutations (amino acids 137 and 147) located in the antigenic domain II of gC, which is responsible for binding of HSV1 virions to the glycosoaminoglycan (GAG) receptor. When expressed in Sf21 cells using the recombinant baculovirus system (Bac-to-Bac), gC(HSZP) and gC(KOS) showed no essential antigenic differences.