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Biomedical subjects

R Rott

Publications and source records attributed to R Rott.

At least 127 records · Page 7Linked to original sources
PubMed1984

Studies on the adaptation of influenza viruses to MDCK cells.

The amino acid sequences and biological properties of the haemagglutinin of three variants of the influenza virus X-31 (H3N2) selected for their capacity to grow in MDCK cells are reported. In two variants, amino acid substitutions at HA1 residues 8 and 144 correlated with the loss of a site for glycosylation and specific changes in antigenicity, respectively. In all three variants substitution of an arginine residue for histidine at HA1 position 17 was correlated with increased pH optima of haemolysis. The importance of this substitution for cleavage of the haemagglutinin precursor required to produce infectious virus is discussed in relation to the three-dimensional structure of X-31 haemagglutinin.

Adaptation, Physiological↗
PubMed1984

Inhibition of proteolytic cleavage of the hemagglutinin of influenza virus by the calcium-specific ionophore A23187.

At calcium-specific ionophore A23187 concentrations of approximately 0.25 microM [which still allow assembly and release of fowl plague virus (FPV) particles] post-translational proteolytic cleavage of the viral hemagglutinin precursor HA into the fragments HA1 and HA2 is inhibited. The resulting virus particles with uncleaved hemagglutinin, that cannot be obtained under normal conditions, provide a suitable substrate for in vitro assays of the protease sensitivity of the FPV hemagglutinin. Proteolytic activation is accomplished with trypsin. Treatment with cathepsin B at low pH yields aberrant cleavage products suggesting that the cellular cleavage enzyme is not of lysosomal origin. A protease that cleaves the FPV hemagglutinin in the correct place can be detected in lysates of MDBK cells. This enzyme is calcium dependent and has a neutral pH optimum.

Animals↗
PubMed1984

Effect of inhibitors of glycosylation on proteolytic activation of avian influenza virus hemagglutinins: discrimination between tryptic cleavage and elimination of the connecting peptide.

The glycosylation inhibitors tunicamycin (TM), 2-deoxyglucose (2-dg), bromoconduritol (BC; 3,5/4,6-6-bromo 3,4,5-trihydroxycyclohex-1-ene), and N-methyl-deoxynojirimycin (MdN) have been used to study the role of glycosylation in the two proteolytic reactions involved in the biological activation of H7 influenza virus hemagglutinins (HAs): trypsinlike cleavage and subsequent elimination of the connecting peptide. The results obtained revealed that trypsin-like cleavage of the HAs of pathogenic strains does not require glycosylation, since these HAs were efficiently cleaved in the presence of TM and 2-dg. The elimination of the connecting peptide between HA1 and HA2, however, appears to require the transfer of oligosaccharides onto the HA polypeptide, since this activity was blocked by TM and by 2-dg. Elimination was not blocked by BC or MdN, which inhibit glucose trimming and subsequent conversion of the high-mannose type to the complex type of carbohydrate.

1-Deoxynojirimycin↗
PubMed1984

On the role of oligosaccharide trimming in the maturation of Sindbis and influenza virus.

The alpha-glucosidase inhibitor bromoconduritol inhibits the formation of the N-linked, complex-type oligosaccharides of the glycoproteins from influenza viruses (fowl plague virus, influenza virus PR-8) and from sindbis virus. Viral glycoproteins produced in bromoconduritol-treated chicken-embryo and baby-hamster kidney cells are fully glycosylated, but accumulate N-linked, high-mannose oligosaccharides of the composition Glc1Manx (GlcNAc)2 (x = 7, 8, and 9). Other alpha-glucosidase inhibitors (nojirimycin, deoxynojirimycin, acarbose) were not specific inhibitors of oligosaccharide processing under the conditions used in the present investigation. In bromoconduritol-treated, sindbis virus-infected chicken-embryo and baby-hamster kidney cells, the sindbis glycoproteins are metabolically stable. Specific proteolytic cleavage of the polyprotein precursors to form E2 and E1 occurs in bromoconduritol-treated chicken-embryo cells, but cleavage of PE2 to E2 is prevented in the infected baby-hamster kidney cells. Yet, release of infectious sindbis virus particles is inhibited in both cell types indicating that the formation of complex oligosaccharides is required for a late step in virus formation. The release of virus particles from influenza virus PR-8-infected bromoconduritol-treated chicken-embryo cells is not inhibited, and virus with only high-mannose oligosaccharides is formed. In contrast, when chicken-embryo cells were infected with the influenza virus fowl plague virus, release of infectious particles was inhibited. The fowl plague virus hemagglutinin is cleaved in chicken-embryo cells, in contrast to the hemagglutinin of the PR-8 virus. However, the cleavage products HA1 and HA2 do not reach the cell surface. In addition, or as a consequence, HA1 and HA2 are proteolytically broken down, whereas uncleaved hemagglutinin of PR-8 appeared metabolically stable. These results may explain the decrease in formation of fowl plague virus particles and the lack of effect on PR-8 virus in bromoconduritol-treated cells. This work thus shows different biological roles for oligosaccharide processing.

Animals↗
PubMed1984

Biological functions of monospecific antibodies to envelope glycoproteins of Newcastle disease virus.

Monospecific antisera to HN and F glycoproteins of Newcastle disease virus were prepared, and their effects on the biological activities of the virus were investigated. Anti-HN serum inhibited hemagglutinating and neuraminidase activity, as well as hemolysis. Anti-F serum had no effect on hemagglutination or neuraminidase but inhibited hemolysis and virus-induced cell fusion. Anti-HN serum was highly neutralizing, while neutralization by anti-F serum was very inefficient in conventional plaque reduction tests, although both sera were estimated to contain comparable amounts of antibody reacting with the virus as indicated by complement fixation and immuno-diffusion tests. The neutralizing activity of anti-F serum was greatly enhanced by the addition of anti-IgG serum or fresh guinea pig serum, whereas that of anti-HN serum was little enhanced. Anti-HN serum incorporated in the agar overlay suppressed the development of plaques to some degree, while anti-F serum had little effect. The combination of anti-HN and anti-F sera resulted in a marked decrease in the number and size of plaques, demonstrating the synergistic effect of the two species of antibody in the containment of the spread of viral infection.

Animals↗
PubMed1984

Replication of Borna disease virus in rats: age-dependent differences in tissue distribution.

There are age-dependent differences in the tissue distribution of Borna disease (BD) virus in rats infected intracerebrally. While in adult rats BD virus replication is restricted to neural cells, in neonatally infected rats infectious virus or viral antigens were found in the cells of most organs. The possibility that differences in the immune status between newborn and adult animals are responsible for different tissue susceptibility could be excluded.

Age Factors↗
PubMed1984

Correlation between loss of the temperature-sensitive phenotype and pathogenicity of fowl plague virus mutants in the chicken.

The reversion of temperature-sensitive (ts) mutants of fowl plague virus to the ts+ phenotype was correlated with pathogenicity for chicken. Two types of ts mutants were investigated: those obtained by mutagenesis with 5-fluorouracil and those obtained by undiluted passages at 33 degrees C. The reversion frequency of the former mutants depended on the RNA segment in which the ts defect was located, mutations in RNA segments 1 and 2 having the highest reversion frequency, those in the RNA segments coding for the glycoproteins the lowest. ts mutants obtained by undiluted passages behaved differently in this respect. There was an approximate correlation between frequency of reversion and pathogenicity for chicken. Double mutants induced by 5-fluorouracil, having one tight and one leaky mutation, reverted easily without loss of the leaky mutation. These double mutants were still to a limited extent pathogenic for the chicken. Only one double mutant with two tight mutations (ts 293) was completely nonpathogenic after intramuscular inoculation. Two ts mutants with multiple tight defects (ts 1/1 and ts 3/18) obtained by undiluted passage did not revert to wild-type after injection into embryonated eggs and incubation at 33 degrees C, but they were still slightly pathogenic for the chicken. There was no obvious correlation between the shut-off temperature and pathogenicity of mutants carrying a single ts defect. However, for mutants with multiple tight mutations a high shut-off temperature seemed to be essential for reversion during serial passages as well as for pathogenicity in the chicken, when different routes of inoculation were examined. ts mutants seem to be safe as live vaccines only, (1) if they carry at least two tight ts defects, (2) if they have a relatively low shut-off temperature, and (3) if they could be administered other than via the respiratory tract.

Animals↗
PubMed1984

Immunogenic properties of the small chain HA2 of the haemagglutinin of influenza viruses.

The small chain of influenza virus haemagglutinin, HA2 was isolated by a selective enzymic removal of HA1 or by preparative SDS-polyacrylamide gel electrophoresis. Anti-HA2 specific antisera and monoclonal antibodies were subtype-specific in immunodiffusion tests and radioimmunoassays. These antibodies did not inhibit haemagglutination or haemolysis, did not prevent virus release, did not neutralize infectivity, and HA2 did not induce a protective immunity. HA2-specific antigenic determinants could not be demonstrated on the surface of infected cells. Lymphocytes from pre-immunized mice could not be stimulated by HA2 to exert a cytotoxic effect.

Animals↗
PubMed1984

[Significance of hemagglutinins for the pathogenicity of avian influenza viruses].

In addition to acute viral diseases, persistent infections have attained considerable interest in recent years. Such persistent infections are characterized by extended time periods in which the infecting virus remains within the organism before the eventual appearance of manifest symptoms. These infections may be evoked by a variety of virus species resulting in a diversity of pathogenic reactions and clinical manifestations. The mechanisms of viral persistence, where known, also appear to be quite diverse. As far as space permits, some examples of persistent infections will be presented and the mechanisms of the pathogenesis of the resulting diseases will be discussed.

Amino Acid Sequence↗
PubMed1983

Behavioral disease in rats caused by immunopathological responses to persistent borna virus in the brain.

Borna virus replicated persistently in the brains of rats, causing frenzied and apathetic behavioral states in sequence but no mortality. The transient frenzied behavior was caused by an immune-mediated, cytolytic, encephalitic response that was unexpectedly self-limiting. Cessation of active pathological processes coincided with the onset of the passive phase of the disease. This study thus demonstrates suppression of virus-specific inflammation despite continuous viral replication and describes a new mechanism by which chronic encephalitis may become established.

Animals↗
PubMed1983

Pathogenicity reactivation of nonpathogenic influenza virus recombinants under von Magnus conditions.

The pathogenicity for the chicken of a number of nonpathogenic recombinants between fowl plague virus and various avian and mammalian influenza A viruses can be reactivated by passaging serially at high multiplicities (von Magnus conditions) at 41 degrees, which is the nonpermissive temperature of nonpathogenic recombinants. While the mechanism underlying this reactivation is unclear, it could be excluded that it was due to segregation of heterozygotes to the wild type homozygotes.

Animals↗
PubMed1983

Evidence for the contribution of the host species to the extent of antigenic variation of N1 influenza virus neuraminidase.

N1 influenza virus neuraminidases (NA) derived from avian, swine and human virus isolates, including the genetically related classic strains A/FPV/Rostock/34, A/Swine/1976/31, A/PR8/34 and A/FM1/47, were analysed serologically by neuraminidase inhibition (NI), inhibition of virus release (IVR) and competitive radio-immunoassays (competitive RIA). Comparing the three tests, competitive RIA appeared to be more reliable than NI and IVR for a quantitative assessment of antigenic relatedness. Together with evidence presented by others, these studies indicate that the host species contributes to the extent of antigenic variation of NAses. In contrast to NAses of human viruses where antigenic drift occurs readily, NAses of animal influenza viruses, from birds or mammalians, undergo far fewer antigen changes.

Animals↗
PubMed1983

Pathogenesis of Borna disease in rats: immune-mediated viral ophthalmoencephalopathy causing blindness and behavioral abnormalities.

Borna disease virus is an unclassified agent that causes a rare but fatal encephalitis in horses in Germany. In experimental animals the virus causes acute fatal encephalitis in some instances and chronic encephalitis with abnormal behavior in others. In initial studies of the pathogenesis of the latter disease in rats, the virus was shown to replicate only in the nervous system, with the greatest concentration of infectivity in the cerebrum and eyes. Viral replication continued indefinitely in both newborn and adult rats. The adult animals developed self-limiting, necrotizing encephalitis in the cerebrum, with inflammation spreading to the retina. Inflammation receded after two months, however, with concomitant cessation of necrosis; static hydrocephalus was observed at this point. Levels of viral replication were unaffected by these changes. Rats became frenzied and aggressive during the encephalitic period but became permanently passive and inactive after inflammation receded. Infected neonates and immunosuppressed adults did not become ill. The frenzied behavior and subsequent blindness in immunocompetent adults were therefore attributed to a uniquely transient immunopathologic reaction targeted to centers in the limbic system and retinal neurons.

Animals↗