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Journal of Virology, May 1999, p. 3843-3853, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Intrastrain Variants of Herpes Simplex Virus Type 1 Isolated from a Neonate with Fatal Disseminated Infection Differ in the ICP34.5 Gene, Glycoprotein Processing, and Neuroinvasiveness

John R. Bower,1,2 Hanwen Mao,1 Catherine Durishin,1 Edgardo Rozenbom,1 Michelle Detwiler,3 Donald Rempinski,3 Tracy L. Karban,3 and Ken S. Rosenthal1,*

Department of Microbiology and Immunology, Northeastern Ohio Universities College of Medicine, Rootstown,1 and Department of Pediatrics, Children's Hospital Medical Center of Akron, Akron,2 Ohio, and Roswell Park Cancer Institute, Buffalo, New York3

Received 13 October 1998/Accepted 21 January 1999

Two intrastrain variants of herpes simplex virus type 1 (HSV-1) were isolated from a newborn with fatal disseminated infection. A small-plaque-producing variant (SP7) was the predominant virus (>99%) in the brain, and a large-plaque-producing variant (LP5) was the predominant virus (>99%) in the lung and gastrointestinal tract. EcoRI and BamHI restriction fragment patterns indicated that SP7 and LP5 are related strains. The large-plaque variants produced plaques similar in size to those produced by HSV-1 KOS. Unlike LP5 or KOS, SP7 was highly cell associated and processing of glycoprotein C and glycoprotein D was limited to precursor forms in infected Vero cells. The large-plaque phenotype from KOS could be transferred into SP7 by cotransfection of plasmids containing the EK or JK EcoRI fragment or a 3-kb plasmid with the UL34.5 gene of HSV-1 KOS together with SP7 DNA. PCR analysis using primers from within the ICP34.5 gene indicated differences for SP7, LP5, and KOS. Sequencing data indicated two sets of deletions in the UL34.5 gene that distinguish SP7 from LP5. Both SP7 and LP5 variants were neurovirulent (lethal following intracranial inoculation of young BALB/c mice); however, the LP5 variant was much less able to cause lethal neuroinvasive disease (footpad inoculation) whereas KOS caused no disease. Passage of SP7 selected for viruses (SLP-5 and SLP-10) which were attenuated for lethal neuroinvasive disease, were not cell-associated, and differed in the UL34.5 gene. UL34.5 from SLP-5 or SLP-10 resembled that of KOS. These findings support a role for UL34.5 in promoting virus egress and for neuroinvasive disease.

* Corresponding author. Mailing address: Northeastern Ohio Universities College of Medicine, P.O. Box 95, Rootstown, OH 44272-0095. Phone: (330) 325-6134. Fax: (330) 325-5914. E-mail: ksr{at}neoucom.edu.

Journal of Virology, May 1999, p. 3843-3853, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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