Ultrastructural Study of Long-Term Measles Infection in Cultures of Hamster Dorsal-Root Ganglion
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Abstract:
The morphogenesis of the Edmonston strain of measles is described in cultures of hamster dorsal-root ganglion maintained for as long as 63 days postinoculation. The patterns observed confirmed those previously reported in both neural and non-neural tissue. However, in the present tissue, the development of viral material could be followed chronologically within different cell types such as neurons and Schwann cells. Active replication was visualized up to 63 days postinoculation. The appearance of cytoplasmic nucleocapsid preceded that of intranuclear nucleocapsid, the latter occurring after 14 days. These intranuclear inclusions were formed after the transformation of the nucleoli into bizarre pleomorphic bodies which eventually segregated into clumps of nucleocapsid. These intranuclear inclusions mimic those seen in subacute sclerosing panencephalitis, now known to be etiologically related to a measles-like virus.Keywords:
Subacute sclerosing panencephalitis
Dorsal root ganglion
Papovavirus
Measles virus generally produces acute illness. Rarely, however, persistent infection of brain cells occurs, resulting in a chronic and fatal neurological disease, subacute sclerosing panencephalitis (SSPE). Evidence indicates that expression of the measles virus matrix protein is selectively restricted in this persistent infection, but the mechanism underlying this restriction has not been identified. Defective translation of matrix messenger RNA has been described in one SSPE cell line. This report presents evidence that in a different SSPE tissue culture cell line IP-3-Ca, the matrix protein is synthesized but fails to accumulate. A general scheme is proposed to reconcile the different levels at which restriction of matrix protein has been observed.
Subacute sclerosing panencephalitis
Morbillivirus
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Subacute sclerosing panencephalitis (SSPE) is characterized by a hyperimmune state toward the polypeptides of measles virus except the matrix (M) protein. Using cloned (3H)-labeled complementary DNA probes for in situ hybridization, we found the IM protein and nucleocapsid (NP) protein nucleotide sequences in glial cells and neurons of cryostat sections from two SSPE brains. In one SSPE brain, M protein was lacking, but the other measles polypeptides were present. IgG and IgM antibodies eluted from that brain lacked antibodies to M protein, but antibodies to other measles polypeptides were present. In SSPE brain, the viral M-protein defect is not a deletion of the M gene, but rather a block in gene expression.
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An appendix removed 15 days before onset of symptoms of subacute sclerosing panencephalitis was examined retrospectively for measles virus ribonucleic acid (RNA). Tissue sections hybridised in situ to a cloned measles virus probe of deoxyribonucleic acid specific for nucleocapsid protein showed that many cells of the lymphoid tissue contained measles virus RNA. In contrast, only a few infected lymphoid cells were detected in three out of six seropositive controls and none in three seronegative infants. A widespread chronic viral infection of the immune system, established after measles, may promote or even initiate nerve cell infection in subacute sclerosing panencephalitis.
Subacute sclerosing panencephalitis
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SUBACUTE sclerosing panencephalitis (SSPE) is a slowly progressing disease of the central nervous system of children and young adults, caused by a persistent measles-virus infection. Patients with SSPE have high levels of antibodies to measles virus in their serum and cerebrospinal fluid (CSF), and the virus has been isolated from the brain; however, isolation requires cocultivation of brain cells with other cultured cell lines.1 2 3 4 5 Measles virus contains six major proteins6 7 8 9: L and P, which are associated with the internal nucleoprotein (nucleocapsid) and are involved in the activity of RNA polymerase; NP, the nucleocapsid structural protein; H, the hemagglutinin, a . . .
Subacute sclerosing panencephalitis
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Morbillivirus
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Subacute sclerosing panencephalitis
Morbillivirus
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Subacute sclerosing panencephalitis
Morbillivirus
Mononegavirales
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The persistence of measles virus in selected areas of the brains of four patients with subacute sclerosing panencephalitis (SSPE) was characterized by immunohistological and biochemical techniques. The five measles virus structural proteins were never simultaneously detectable in any of the brain sections. Nucleocapsid proteins and phosphoproteins were found in every diseased brain area, whereas hemagglutinin protein was detected in two cases, fusion protein was detected in three cases, and matrix protein was detected in only one case. Also, it could be shown that the amounts of measles virus RNA in the brains differed from patient to patient and in the different regions investigated. In all patients, plus-strand RNAs specific for these five viral genes could be detected. However, the amounts of fusion and hemagglutinin mRNAs were low compared with the amounts in lytically infected cells. The presence of particular measles virus RNAs in SSPE-infected brains did not always correlate with mRNA activity. In in vitro translations, the matrix protein was produced in only one case, and the hemagglutinin protein was produced in none. These results indicate that measles virus persistence in SSPE is correlated with different defects of several genes which probably prevent assembly of viral particles in SSPE-infected brain tissue.
Subacute sclerosing panencephalitis
Morbillivirus
Mononegavirales
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Comparative studies between two measles virus strains isolated from patients with subacute sclerosing panencephalitis (SSPE) and a prototype low tissue culture passage Edmonston measles virus are described. Differences were noted in several properties. The findings described in this report suggest that strains of measles virus associated with SSPE have different biological properties and apparently cannot be distinguished from laboratory and field strains of the virus.
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Morbillivirus
Mononegavirales
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Groups of oligoclonal immunoglobulin G (IgG) bands were isolated from sera of patients with subacute sclerosing panencephalitis by employing preparative isoelectric focusing. Six IgG fractions containing two to three oligoclonal bands with different isoelectric points were used to precipitate the proteins from Vero cells infected with measles virus. The results showed that all of the measles virus proteins except the M protein were precipitated by all of the IgG fractions and that the precipitation of viral proteins by the fractions containing groups of oligoclonal IgG showed slightly different patterns in some sera, whereas other sera showed no significant differences. The present study indicates that oligoclonal IgGs in subacute sclerosing panencephalitis sera are not specific to individual measles virus proteins.
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Measles virus was isolated from brain cell tissue cultures derived from two SSPE patients. These cultures proved to contain intracellular measles antigen which was not released in the fluid phase. Infectious, complete virus was obtained when mixed cultures containing the brain cells and HeLa cells were prepared. It appears that SSPE is due to suppressed measles virus infection. Once “rescued” through the mixed culture technique, the virus recovered from SSPE patients proved indistinguishable from measles virus.
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