Nucleotide sequences responsible for the inability of a herpes simplex virus type 2 strain to grow in human lymphocytes are identical to those responsible for its inability to grow in mouse tissues following ocular infection.
A study was undertaken to determine whether genes associated with herpes simplex virus (HSV) neuroinvasiveness in mice influence the growth of HSV in man, the virus's natural host. HSV-2(186), a nonneuroinvasive HSV strain, was found to replicate poorly (less than 3-fold) in cultures of phytohemagglutinin (PHA) stimulated human peripheral blood mononuclear cells (PBMC). In contrast, seven other HSV strains all multiplied 40- to 100-fold. The paucity of HSV-2(186) growth in PBMC was not due to a failure of this strain to grow in primary human cells because high titers (greater than 10(8) PFU/ml) were obtained following infection of human foreskin fibroblasts. The genetic basis for the deficient growth was analyzed by marker rescue experiments. Recombinant HSV-2 strains were generated in marker rescue experiments utilizing HSV-2(186) DNA and plasmids containing a cloned DNA polymerase gene isolated from a neuroinvasive HSV strain possessing the capacity to replicate in human PBMC. Progeny which rescued DNA from the cloned HSV DNA polymerase gene replicated 40- to 100-fold in PHA-stimulated PBMC. Moreover, unlike the HSV-2(186) parent, HSV-2(186) isolates possessing rescued DNA grew well in the eye, trigeminal ganglion, and brain of mice and induced fatal encephalitis. The results indicate that nucleotide sequences responsible for increasing the capacity of HSV-2(186) to grow in PBMC of man are identical to those responsible for increasing the capacity of this strain to grow in mouse tissues and to spread from the eye to the brain.
Lausch
R N
RN
Department of Microbiology/Immunology, College of Medicine, University of South Alabama, Mobile 36688.
Yeung
K C
KC
Miller
J Z
JZ
Oakes
J E
JE
D000328:Adult; D000818:Animals; D015139:Blotting, Southern; D001921:Brain / Q000382:microbiology; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D015245:Deoxyribonuclease BamHI; D004660:Encephalitis / Q000209:etiology; D005123:Eye / Q000382:microbiology; D005347:Fibroblasts / Q000382:microbiology; D005819:Genetic Markers; D006801:Humans; D007635:Keratitis, Dendritic / Q000150:complications / Q000382:microbiology*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000382:microbiology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D015183:Restriction Mapping; D018139:Simplexvirus / Q000235:genetics* / Q000502:physiology; D012668:Trigeminal Ganglion / Q000382:microbiology; D014709:Vero Cells; D014779:Virus Replication / Q000235:genetics
D004279:DNA, Viral; D005819:Genetic Markers; D004259:DNA-Directed DNA Polymerase; D015245:Deoxyribonuclease BamHI