Persistent poliovirus infection of human neuroblastoma cells.

Abstract

Two human neuroblastoma cell lines were persistently infected with poliovirus strains of all three serotypes. In persistently infected IMR-32 cells, which were studied in greatest detail, viral antigens were present in most cells, and over a 9-month period virions were found in the medium at high titers. Persistently infected cells were resistant to superinfection by Sabin 1, 2, and 3 poliovirus but sensitive to coxsackievirus B3. The viruses recovered from persistently infected cells were studied for conservation of epitopes, host cell specificity, and temperature resistance phenotype. The antigenic site 1 carried by the major capsid protein VP1 was modified on the persistent viruses of all three serotypes. This was confirmed for one virus by sequencing the corresponding genomic region in which two mutations were detected. The titers of persistent viruses were 1-3 log10 units higher on IMR-32 cells than on nonneuronal HEp-2 cells, while parental viruses had similar titers on both lines. When thermosensitive viruses were used to initiate the infection, the persistent viruses were found to be thermoresistant at 390C. Together the results indicate that the persistent infection correlated with the selection of highly mutated viral strains. Poliovirus-infected neuroblastoma cell lines thus constitute an in vitro model ofchronic viral infections, which are increasingly implicated in human neural diseases.Poliomyelitis paralyses are caused by poliovirus (PV)-induced necroses of motor neurons (1, 2). A late postpolio syndrome has been described: new focal motor neuron deterioration emerges after an average period of 30 years from initial infection (3, 4). One of the hypotheses proposed to account for this syndrome is a persistent viral infection (3, 4). Several members ofthe picornavirus family are effectively able to induce a persistent infection ofcells in vivo and in vitro (5-11). It is assumed that the selection of viral mutants, viral interference, and interferons play a role in this persistence (12,13). For PV, the viral cytolytic action was shown to be limited by the intercellular matrix (14). Cultures in which only a small fraction of cells were susceptible to PV have been described (15)(16)(17). In a more recent study, Kaplan et al. (18) isolated PV-infected HeLa cell lines blocked at different steps of the virus life cycle which underwent periodic crises with cytopathic effects (CPE).In cell lines of neuronal origin, PV-cell interactions have been studied only in one-step growth experiments during the first 8 hr post infection (p.i.) (19). We report here the establishment of a persistent PV infection of human neuroblastoma cells. Most cells harbored viral antigens and cultures produced virions for months at high titers in the absence of detectable CPE. Persistently infected cell lines and the resulting PV mutants are described.MATERIALS AND METHODS Materials. The Leon 37, vFG68 (20), LSc2ab (S1), P712 Ch 2ab (S2), and P3/Leon 12alb (S3) Sabin strains (21) of PV were used. Another enterovirus, coxsacki...

Transgenic expression of replication-restricted enteroviral genomes in heart muscle induces defective excitation-contraction coupling and dilated cardiomyopathy.

“…Several picornaviruses including CVB3 have been shown to establish persistent infections in vivo and in vitro in several diseases and cell types (5)(6)(7)(8)(9)(10)(11)(12)(13)(14). Persistent Coxsackieviral infection in the murine heart is qualitatively distinct from acute viral infection.…”

Topology of Double-Membraned Vesicles and the Opportunity for Non-Lytic Release of Cytoplasm

“…However, the possibility of nonlytic release of poliovirus and other picornaviruses has been suggested from numerous reports of persistently infected cell lines that continuously secrete infectious particles. 17,18,14,19 Even more convincingly, when polarized Caco-2 cultures, growing as intact monolayers, were infected with poliovirus, newly synthesized virus was shown to emerge only from the apical surface. 14 Assays showed that the monolayer remained intact, arguing that a non-lytic, and polarized, exit route of unknown origin had been utilized.…”

Complete Nucleotide Sequence of a Coxsackievirus B4 Strain that Establishes Infection in ICR Mice Pancreas and Induces Glucose Intolerance

“…Other researchers have reported that the coxsackie B virus could also use the decay accelerating factor (DAF, CD55; Bergelson et al, 1995;Shafren et al, 1995), a 70-kDa protein that protects cells from complement-mediated lysis. Early studies revealed that one of the most important factors determining the ability of these viruses to establish a persistent infection in vitro is the change of receptor usage compared with their parental strains or with the prototype strains (Colbere-Garapin et al, 1989;Zhang and Racaniello, 1997;Duncan et al, 1998). This change of receptor usage has been shown to be because of amino acid substitutions in the capsid proteins (Duncan and Colbere-Garapin, 1999;Schmidtke et al, 2000).…”