Restriction Endonuclease Analysis of Herpes Simplex Virus from Recrudescent Lesions, from Latent Infection and During Passage in the Skin and Nervous System of Mice

Abstract

SUMMARYThe stability of the restriction endonuclease profile of herpes simplex virus type 1 strain SC16 in mice was studied. Virus isolated from skin during acute infection was compared with that from latently infected ganglia and with that from recrudescent lesions induced by trauma. In another experiment virus serially passaged only in skin was compared with virus that had also replicated in the nervous system. The loss or gain of specific restriction sites was not observed but in some cases the mobility of certain fragments decreased.Restriction endonuclease analysis of the viral DNA can clearly differentiate between the two types of herpes simplex virus (HSV) (Skare et al., 1975) and distinguish many intratypic variants (Buchman et al., 1978). Almost every clinical isolate from different patients has its own distinct profile (Buchman et al., 1978), except in cases of proven or likely contact (Buchman et al., 1978; J. M. Darville, unpublished observations). However, despite this variety of profiles, those obtained from successive isolates from the same individual show at most only minor differences in the mobility of some fragments (Maitland et al., 1982; J. M. Darville, unpublished observations). This suggests that within one individual the virus retains a constant genetic pattern over long periods and raises the question of the origin of the diversity between strains from different individuals. In this study we have attempted to find by endonuclease analysis whether changes in the viral DNA occur in mice experimentally infected with HSV-1. We investigated the restriction profiles of the DNA from virus isolated from primary infection, recrudescent lesions and latently infected ganglia. In addition, we examined DNA from virus passaged repeatedly only in the skin and from that isolated at each passage from the nervous system. Restriction profiles of the DNA of virus passaged repeatedly in cell culture were also examined.The virus used was HSV-1 strain SC16 (Hill et al., 1975), isolated from a human recurrent lesion and passaged five times in Vero cells. To preserve any heterogeneity in the original population the virus was not cloned. Mice were female (to avoid damage to the ears from fighting) outbred Swiss white (Hill et al., 1975); they were 4 weeks old when inoculated.All HSV isolates were subjected to restriction enzyme analysis by a simplified method (Darville, 1983) derived from one previously described (Lonsdale, 1979). In the first experiment the enzymes used were BstI, EcoRI (Cambridge Biotechnology Laboratories, CBL), PvulI and KpnI (Bethesda Research Laboratories). In later experiments EcoRI and KpnI were replaced by SstI (CBL) since HSV has more sites recognized by this enzyme.In the first two experiments samples were so coded before analysis that all isolates from an animal could be examined blind in the correct order in adjacent tracks of the same gel.The first experiment was designed to compare virus replicating at the inoculation site during primary infection with that isolated from latentl...

Possible latent infection with herpes simplex virus in the mouse eye

“…The work of Subbuga et al (1988) demonstrates that HSV nucleic acid can persist in the ocular tissue of experimental animals when the primary infection has subsided and this could represent HSV in a latent state. Our findings that the isolates of HSV from cultured AS were identical to the strain used for the original inoculation as defined by restriction endonuclease analysis (data not shown) are in agreement with those of Darville et al (1987) who studied virus strains isolated from recrudescent lesions in the skin. They also agree with those of Caudill et al (1986), although in the latter experiments (where HSV shedding was activated by iontophoresis) the source of virus could equally have been from the latently infected TG.…”

Experimental in vivo generation of intertypic recombinant strains of HSV in the mouse

“…The restriction enzyme profile of LIP 5 C remained constant during primary infection and other strains of HSV have shown similar stability during acute and latent infection in both mice [3,17] and humans [13]. Therefore, the variants observed here are most likely to be recombinants.…”

“…1 HSV-1/LIP 5 C (PFAr). [2][3][4] Recombinants from skin of inoculation site during acute infection (designated "type 1"). 5-8, 10, 12 Recombinants from ganglia during acute infection (7 "type 2", others "type 1").…”

Reiterated sequences of herpes simplex virus type 1 (HSV-1) genome can serve as physical markers for the differentiation of HSV-1 strains

“…Thus, the common type variation can be detected more easily. Second, the reiterated sequences in common type variations have a tendency to be more variable than RFLP [6,7,13,19,21,22,[29][30][31]. The lengths of reiterations are assumed to be readily changed by crossover events between tandemly repeated units on the 296 K. Umene and M. Yoshida reiterations.…”