Comparative pathology of infections with baboon and African green monkey alpha-herpesviruses in mice.

The comparative pathology of Herpesvirus papio 2 (HVP2) of baboons and SA8 virus of African green monkeys relative to that of herpes simplex virus (HSV1) of man was investigated in young adult mice inoculated intramuscularly and observed for 21 days. The 50% infectious dose (ID(50)) for HVP2 was approximately 10(2.0) plaque-forming units (PFU), while the ID(50) for HSV1 and SA8 was 10(2.5) and 10(3.8), respectively. There were marked differences in the ability of these three viruses to invade the central nervous system (CNS) and cause clinical neurological disease. HSV1 produced neurological signs in a few animals given 10(6)PFU, but SA8 did not. In contrast, HVP2 readily invaded the CNS and produced fatal disease with doses as low as 10(2)PFU. Two isolates of HVP2 tested had a 50% CNS disease dose (CNSD(50)) of 10(2.5) and 10(3.0)PFU and an LD(50) of 10(3.8) and 10(4.3)PFU, respectively. Histopathological examination of tissue from HVP2-infected mice revealed severe lesions of inflammation and necrosis in the central, peripheral and autonomic nervous systems, as well as of other tissues including skin, adrenal glands and the gastrointestinal tract. Viral antigens were detected immunohistochemically in lesions. This study showed that while both HVP2 and SA8 could infect mice, there were marked differences in the ability of these two closely related viruses to cause clinical disease and CNS lesions. This murine model may prove useful in the investigation of viral or host determinants responsible for the varying neurovirulence of these simian alpha-herpesviruses.

Identification of a 3'-->5'-exonuclease that removes cytosine arabinoside monophosphate from 3' termini of DNA.

Cytosine arabinoside monophosphate (araCMP) at the 3' terminus of DNA constitutes a lesion that impedes further synthesis by DNA polymerase alpha (DNA pol alpha). A biochemical assay has been designed to detect 3'-->5'-exonucleases in cell extracts that remove the 3'-araCMP lesion in an oligonucleotide template-primer and permit subsequent extension by DNA pol alpha. The major 3'-->5'-exonuclease activity in human myeloblast extracts has been purified, and gel filtration chromatography of the purified enzyme indicates that the exonuclease has an apparent native molecular mass of 52 kDa. Incubation of the enzyme with a 5'-32P-labeled araCMP template-primer results in exonucleolytic degradation of the primer exclusively in the 3'-->5' direction, demonstrating that the enzyme is a 3'-->5'-exonuclease. The products of the 3'-->5'-exonuclease reaction are 5'-mononucleotides. The apparent rate of araCMP removal by the exonuclease is approximately the same as the rate of deoxynucleoside monophosphate (dNMP) removal. Furthermore, the apparent rates of 3'-terminal excision are approximately the same whether the oligomer is hybridized to a complementary oligonucleotide, or not, indicating that the enzyme has both single- and double-stranded 3'-->5'-exonuclease activity. The enzyme does not possess 5'-->3'-exonuclease activity, nor is it associated with DNA polymerase activity. In addition, the enzyme does not cleave 3'-phosphoryl-terminated DNA, and it does not cleave RNA. The enzymatic characteristics of the isolated 3'-->5'-exonuclease indicate that it is distinct from previously identified mammalian deoxyribonucleases.