Yuzo Aoyama

Pathology of the human spinal ganglia in varicella-zoster virus infection.

SummarySpinal ganglia from a patient who died on the 6th day of varicella infection were examined by immunofluorescence and electron microscopy, and were compared with spinal ganglia from a patient dying on the 17th day of herpes zoster infection. In herpes zoster, typical intranuclear inclusion bodies were found in neurons, satellite cells and fibroblast-like cells of the ganglia, which contained numerous naked virus particles. In varicella, few changes were found by light microscopy but viral antigen was detected in a few neurons and satellite cells by immunofluorescence. Electron microscopy revealed scattered virus particles near the nuclear membrane of a neuron, satellite cells and capsular cells and enveloped particles in the cytoplasma of satellite cells. The particles in the nuclei were mostly naked virions with specific crescent-like inner-nuclear structure; those in the cytoplasm had complete and incomplete envelopes and showed pleomorphism. A “virus-like” intranuclear filament found in mononuclear cells in herpes zoster and a “plexiform vermicellar array” found in the nuclei of neurons in varicella are at present considered to be non-specific nuclear changes caused probably by viral infections.

Detection of Viral Antigens in Formalin‐fixed Specimens by Enzyme Treatment

Enzyme treatment (protease or trypsin) was applied to formalin-fixed paraffin-embedded materials and virus-infected cultured cells to detect viral antigens by immunofluorescence. The viral antigens were demonstrated in several organs of autopsy or biopsy cases of which diagnoses had been established by immunofluorescence or virus isolation using frozen materials, or suspected on the basis of serology and/or histopathological findings. These included herpes simplex, varicella-zoster, cytomegalo, subacute sclerosing panencephalitis, progressive multifocal leukoencephalopathy, Japanese B encephalitis, measles, acute hemorrhagic conjunctivitis, Lassa and Korean hemorrhagic fever. Antigen could be recovered also in virus-infected cells (herpes simplex, measles, Lassa, Ebola, Marburg, Rift Valley, Congo and Korean Hemorrhagic fever) by enzyme treatment after periods of formalin fixation of four weeks and storage of three months. In herpes simplex virus-infected mouse brain, antigen was detected after fixation for three months in formalin.

Comparative studies of several vaccinia virus strains by intrathalamic inoculation into cynomolgus monkeys.

SummaryFrom the comparative studies of the virulence of several vaccinia virus strains by intrathalamic inoculation into cynomolgus monkeys, the following results were observed. The CV1 virus was most virulent, the New York City Board of Health, Ikeda, EM63, and Lister viruses were slightly less virulent, and DIs and LC 16 viruses least virulent.The characteristic findings were widespread inflammatory lesions in the meninges and choroid plexus which were closely associated with the replication of vaccinia virus, and parenchymal lesions which might be referred to as encephalopathy in the deceased monkeys. Meningoencephalitis was, however, often recognized in the monkeys sacrificed at 14 days postinoculation and those dying late.

Varicella virus isolation from spinal ganglion

There is strong support for the view that herpes zoster of skin represents a reactivation of varicella virus which has remained latent in one or more of the spinal or cranial ganglia following an episode of varicella infection (5, 6). However, information regarding the presence of virus in nervous tissues is extremely meager despite the regular demonstration of virus in the skin lesions. The present paper describes the isolation of varicella virus from a zoster patient not only from the skin lesion but also from the corresponding spinal ganglion which had typical histological lesions and varicella antigens demonstrable by immunofluorescence. A 71-year-old Japanese female developed vesicles of herpes zoster on the right chest 3 days before her death due to bacterial pneumonia. Autopsy revealed pneumonia in the left lung and the right lower lobe. Multiple fresh vesicles covered an area of the right lateral chest corresponding roughly to the 7th and 8th thoracic dermatomes. The 7th, 8th and 9th thoracic ganglia on both sides, the corresponding segments of the spinal cord and the affected skin region were removed at autopsy. The skin specimen was then subjected to virus isolation. The other specimens were cut into 2 portions, one for virus isolation and the other for histological and immunofluorescent examinations, and stored at 8 0 ~ in Eagles minimum essential medium (MEM) containing 10 per cent fetal calf serum (FCS) and 7 per cent dimethylsulfoxide until use. The conventional histological examination revealed tha t the right 7th thoracic ganglion was severely affected, while the other ganglia and the spinal cord showed no changes. The central portion of the affected ganglion showed complete hemorrhagic necrosis, which was surrounded by a less damaged zone showing degeneration of ganglion cells, satellite cells, Schwann cells and nerve fibers, accompanied by infiltration of lymphocytes and polymorphonuelear leukoeytes and congestion of blood vessels. Ghosts of ganglion cells were identified on account

Analysis of viral antigens in giant cells of measles pneumonia by immunoperoxidase method.

The localization of measles virus proteins was analyzed by immunoperoxidase method using both monospecific and monoclonal antibodies. In Vero cells infected with the Edmonston or EB-L strain, the former being a laboratory strain and the latter a fresh isolate from a measles patient, nucleocapsid protein was located in the nuclei, and matrix protein, phosphoprotein, haemagglutinin and fusion protein were located in the cytoplasm. In the lung tissues of eight cases with measles giant cell pneumonia, the similar findings were obtained. The presence of haemagglutinin on the surface of giant cells at the luminal side was also noticed. Histopathologically, measles giant cells had nuclear and cytoplasmic eosinophilic inclusion bodies with some differences in appearance. The significance of localization of viral proteins is discussed in comparison with histopathological findings in measles giant cells.

Genome variation among varicella-zoster virus isolates derived from different individuals and from the same individuals

SummaryWe have used 12 restriction enzymes to analyse the DNA of 24 clinical isolates of VZV derived from 12 patients in order to compare isolates derived from different individuals and derived serially from the same individual. As reported previously, only a small proportion of the isolates differed with respect to the presence or absence of restriction sites. However, we found that the size of DNA fragments generated from all the isolates derived from different patients varied in any of four regions, one of which was first recognized in this study. In one case, where multiple isolates recovered from the same individual were analysed, each was distinguished from the others not only by differences in the variable regions but also by the presence or absence of a restriction site in a nonvariable region. This suggests that multiple strains of VZV can be present in the same human host.

A Neurotropic Variant of Measles Virus in Suckling Mice.

Strain Sugiyama, adapted to FL cells after 6 passages in primary monkey renal cells, acquired the ability to propagate serially in the brain of suckling mice somewhere between the 49th and 76th FL passage. The virus produced a lethal spastic paralysis in about half of mice at the early passages and in almost 100% from the 9th passage on. Mice were nearly 100% susceptible within 5 days after birth and almost 100% resistant at the age of 7 days or over. Extraneural routes of inoculation were ineffective for infection. Histologic changes were found only in the cerebrum, not in the cerebellum or spinal cord, nor in other viscera organs, and consisted of degeneration of nerve cells with or without formation of inclusions characteristic of measles, multinuclear giant cell formation by fusion of nerve cells, and proliferation of glia cells. Fluorescent antibody study revealed remarkable specificity of nerve cell involvement. Specific measles antigens were demonstrated predominantly in nerve cells, only rarely in glia cells, but not in other types of cell.

THE DISTRIBUTION AND LOCALIZATION OF IMMUNO‐GLOBULIN IN THE GASTRIC MUCOSA AND GASTRIC CANCER

The distribution and localization of immunoglobulin in the gastric mucosa and gastric cancer were studied by means of immunofluorescence. In the lamina propria of the stomach, like the other parts of the gastrointestinal tract, IgA producina cells predominated, but their population density varied from case to case. The main portion of secretion of IgA in the stomach was deep foveolar epithelium. IgA containing epithelial cells in the stomach were much less than those in the intestine, but they increased with advancement of intestinal metaplasia. Some gastric cancer cells retained the ability of secretory component production and took up IgA in their cytoplasm. ACTA PATH. JAP. 29: 523–531, 1979.

ADENOVIRUS HEPATITIS IN A PATIENT WITH SEVERE COMBINED IMMUNODEFICIENCY

An autopsy case of adenovirus hepatitis with severe combined immunodeficiency (SCID) is presented. A 7‐month‐old female was admitted to the Kitasato University Hospital with chief complaint of persistent fever. A diagnosis of severe combined immunodeficiency (SCID) was made because of defective function in both T and B lymphocytes. Respiratory disturbance and severe hepatic dysfunction were manifested after admission. She died of respiratory failure on the 40th hospital day. Autopsy findings revealed many focal necroses scattered irregularly throughout the liver. Degenerating he‐patocytes around the focal necrosis contained nuclear inclusion bodies, and crystalline arrays of adenovirus virions were recognized in these cells by electron microscopy. Adenovirus antigens were brilliantly detected in the nuclear inclusion bodies by indirect immunofluorescence.

Measles associated with coronary arteritis

A two-year-old girl with measles virus (MV) and chronic Epstein-Barr virus (EBV) infection developed lethal coronary aneurysmal arteritis accompanied by giant cell pneumonia, systemic lymphadenitis and hepatosplenomegaly. In her coronary arteries, lungs and aorta, cells containing intranuclear and intracytoplasmic inclusions, including syncytial giant cells, were detected, the presence of MV in the organs being proved by electron microscopic and immunofluorescent studies. Immunopathology further demonstrated MV to be disseminated in almost all organs other than lymph nodes. Clinical diagnosis of chronic EBV infection was established on the basis of persistent high titers of antibodies against capsid and early antigens of EBV and viral presence was confirmed by Southern blot hybridization in a mesenterial lymph node obtained at autopsy. To the best of our knowledge, this is the first description of MV association with coronary aneurysmal arteritis, raising the possibility that measles infection can cause severe vasculitis under immuno-suppressive states, such as that caused by chronic EBV infection.