Margaret Lenahan

Identification of herpes simplex virus infection by immunoperoxidase and in situ hybridization methods

Seven cases of visceral herpes simplex virus (HSV) infection were observed in five cases of hematopoietic disease and in one case each of a newborn baby and a pregnant woman. These seven cases were studied with an immunoperoxidase method and in situ hybridization. In HSV lesions of squamous epithelium, the immunoperoxidase method using rabbit anti-human HSV revealed positive staining, mainly in the nucleus but with some cytoplasmic staining. DNA in situ hybridization revealed stronger positive staining in the nucleus. In HSV hepatitis positive staining was seen in the nucleus and cytoplasm, both by immunoperoxidase and in situ hybridization methods. In the newborn baby, HSV lesions were observed in the brain only, with numerous positive astrocytes identified by the immunoperoxidase method and a few positive astrocyte nuclei by in situ hybridization. Cultured human fetal fibroblasts from the lung were infected with HSV. The immunoperoxidase method revealed diffuse positive staining in the nucleus and in the cytoplasm whereas in situ hybridization revealed fibrillar positive staining in the nucleus only. Thus, the immunoperoxidase method using rabbit antihuman HSV can detect the presence of HSV protein more sensitively than in situ hybridization, probably because of the greater quantity of HSV protein compared with HSV DNA in infected cells.

The pathogenesis of poliomyelitis. Sites of multiplication of poliovirus in cynomolgus monkeys after alimentary infection

The P 1549 (antigenic type 1) strain of poliovirus was administered by gavage to cynomolgus monkeys in order to obtain additional information on primary sites of multiplication of poliovirus. The first sites of virus multiplication were in tissues of the alimentary tract, principally in the oropharynx, and possibly in the ileum. During the early period in pathogenesis poliovirus was found also in lymph nodes collateral to these alimentary sites of multiplication. As early as 24 hours after detection of poliovirus in these tissues low concentrations of virus were detected in the blood; the concentration of virus in the blood rose gradually to reach maximal levels within 48 to 96 hours; thereafter, the concentration gradually fell until no further evidence of viremia was obtained by the 10 th day after infection. When the viremia was approaching, or had reached maximal levels poliovirus was distributed widely in all 46 tissues examined for its presence. Near the end of the period of viremia specific neutralizing antibodies were also found in the blood.

Propagation of Group A Coxsackie Viruses in Primary Human Amnion Cells. I. Cytopathic Changes Produced by 5 More Serotypes.

Summary Seventeen serotypic Group A Coxsackie viruses, representing prototypes established by Dalldorf and Sickles(5,10) have been propagated in tissue cultures. Sixteen strains produced CPE in primary human amnion cells; another (type 7) which has failed thus far to affect amnion cells produced CPE in renal cells obtained from monkeys. Among these 17 Group A viruses were 5 serotypes heretofore not associated with CPE in tissue cultures.

Identification of cytomegalovirus infection in acquired immunodeficiency syndrome

Cytomegalovirus (CMV) infection was observed in 10 of 12 autopsy cases of acquired immunodeficiency syndrome (AIDS) and appears to be the commonest life-threatening viral infection in AIDS. In all 10 cases, adrenal glands were affected with CMV and adrenal medullary necrosis was present in 6 cases. Lungs were affected with CMV in 7 cases with disseminated infection and positive CMV culture. In situ hybridization of tissue sections with CMV-specific DNA provided positive staining for CMV in inclusions as well as other infected cells without obvious inclusions. Human diploid lung fibroblasts were infected with isolated CMV in culture, yielding positive CMV identification within 5 days by in situ hybridization before specific cytopathic changes appeared in the fibroblasts. The early and specific detection of CMV is possible by in situ hybridization with cultured fibroblasts.