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C Orvell

Publications and source records attributed to C Orvell.

At least 73 records · Page 4Linked to original sources
PubMed1991

A single-season epidemic with respiratory syncytial virus subgroup B2 during 10 epidemic years, 1978 to 1988.

Respiratory syncytial virus strains isolated from infants and children hospitalized with acute respiratory tract disease in Huntington, W. Va., from July 1979 through June 1988 were categorized by subgroup by using two panels of monoclonal antibodies specific for subgroup A and subgroup B. Of 405 strains tested, 319 (78.8%) were subgroup A, 73 (18.0%) were subgroup B1, and 13 (3.2%) were subgroup B2. The subgroup B2 strains occurred as a single-season epidemic during the epidemiologic year July 1980 to June 1981, except for one strain isolated in the epidemiologic year 1978 to 1979. Subgroup B strains were associated with significantly fewer bronchiolitis illnesses than subgroup A strains. The severity of illnesses caused by subgroup B1 and B2 strains was the same.

Age Factors↗
PubMed1990

Comparison of two morbilliviruses isolated from seals during outbreaks of distemper in north west Europe and Siberia.

Recently morbilliviruses were isolated from harbour seals (Phoca vitulina) in North West Europe (phocid distemper virus-1: PDV-1) and from Baikal seals (Phoca sibirica) in Siberia (phocid distemper virus-2: PDV-2) during outbreaks of severe disease which resembled distemper in dogs. PDV-1 and PDV-2 were passaged in SPF dogs, in which they caused distemper-like disease symptoms, and were subsequently passaged in Vero cells in which they caused cytopathic changes. PDV-1, PDV-2, and canine distemper virus (CDV) were compared with respect to their biological, morphological, physical, protein chemical, and antigenic properties. It was concluded that PDV-1 should be considered a newly recognized member of the genus Morbillivirus, whereas PDV-2 proved to be quite similar if not identical to CDV.

Animals↗
PubMed1990

Hemagglutinin-neuraminidase (HN) amino acid alterations in neutralization escape mutants of Kilham mumps virus.

The hemagglutinin-neuraminidase genes of the Kilham strain of mumps virus and three neutralization escape mutants (M11, M12 and M13) of this strain (Löve et al., 1985a) were sequenced using their genomes as template. The predicted amino acid sequences were compared. While one mutant had only one amino acid substitution the other two mutants had four and five respectively. A putative region for the epitope of the selected neutralizing monoclonal antibody was identified in a hydrophilic region encompassing amino acids 352-360, since the single amino acid substitution of one mutant occurred in this region and the other two mutants showed non-conserved amino acid changes in this part of the protein. The previously sequenced prototype strain RW, which lacks capacity to react with the selected neutralizing monoclonal antibody also has one non-conserved amino acid change in the region of the proposed neutralizing epitope. The three mutants showed different biological characteristics. These particular characteristics were therefore interpreted to be primarily associated with strain-specific amino acid changes outside the region of the presumed neutralizing epitope. The decrease in molecular weight in one mutant (M11) was shown to be due to a substitution in position 329 of an asparagine for an aspartic acid, leading to abolishment of a potential N-linked glycosylation site. In the other mutants, one substitution in position 239 of a lysine for a methionine was correlated with an increased neuraminidase activity of strain M12, while a substitution in position 360 of an arginine for a cysteine appeared to represent the most likely explanation for the reduced neurovirulence of strain M13.

Amino Acid Sequence↗
PubMed1990

Virus and host cell-dependent variation in transcription of the mumps virus genome.

Evidence has been presented that generation of polycistronic readthrough RNAs in mumps virus-infected cells is not a simple stochastic process with strain-dependent variations in the generation of certain readthrough products, but that this process is affected by host as well as viral factors. RNAs extracted from infected Vero cells or chicken embryo fibroblast (CEF) cells have been analysed by Northern blotting with virus-specific probes for the nucleocapsid (N), phosphoprotein (P), matrix (M), fusion (F), small hydrophobic (SH) and haemagglutinin-neuraminidase (HN) genes. Vero cells infected with tissue culture cell-adapted virus strains generate monocistronic as well as polycistronic RNAs. Transcription analysis of Vero cells infected with an egg-adapted strain reveal the absence of monocistronic M and F transcripts, with a concomitant increase in readthrough transcripts involving these genes. When the same virus infects CEF cells monocistronic RNAs accumulate. The presence of viral proteins in the various virus/host cell combinations assessed by immunofluorescence with mumps virus-specific monoclonal antibodies for the N, P, M, F and HN proteins correlates well with the patterns of transcription.

Animals↗
PubMed1990

Immunological relationships between phocid and canine distemper virus studied with monoclonal antibodies.

The immunological relationships between distemper viruses, isolated from a seal and mink in Denmark and from a dog in Greenland, were investigated with 39 previously developed monoclonal antibodies (MAbs) directed against four major structural proteins of canine distemper virus (CDV). They were also investigated with 16 newly developed MAbs directed against the fusion (F) and large glycoprotein (named H in analogy with measles virus) of phocid distemper virus (PDV) isolated from a harbour seal (Phoca vitulina). These MAbs were reacted with the three different isolated viruses and with the LEC strain of measles virus, in ELISA and immunofluorescence tests. In addition, immunoprecipitation tests were carried out with some of the cross-reacting antibodies. All 55 MAbs reacted identically with distemper virus isolated from seals or mink. When the MAbs produced against CDV were tested, 37 of 39 antibodies reacted with a virus isolated from a sled dog diseased in an outbreak of distemper in Greenland prior to the epizootic among seals in the North Sea. Of the 39 antibodies, 25 reacted with PDV and distemper virus isolated from mink. Of these antibodies, only three of the nine antibodies directed against the H protein of CDV cross-reacted with PDV and distemper virus from mink. Eleven MAbs, reacting with six epitopes of the H protein of PDV, were produced. All 11 antibodies reacted with distemper virus from mink, two of the antibodies reacted with CDV and none reacted with measles virus. All five antibodies reacting with three different epitopes of the F protein of PDV reacted with distemper virus from mink and CDV. Of these five antibodies three, directed against two epitopes, reacted with measles virus. Of the two envelope proteins, the H protein shows pronounced immunological differences between PDV and CDV. In contrast, immunologically the F protein appears to be well conserved among morbilliviruses. It is concluded that the virus causing the epizootic in seals in the North Sea in 1988 may have infected mink on land, or, alternatively, the virus in the sea may have originated from virus-infected mink.

Animals↗
PubMed1990

Segregation of viral structural proteins in cultured neurons of rat spinal ganglia and cord.

Cultured spinal ganglion and spinal cord neurons were used to examine the intraneuronal distribution of five structural proteins of Sendai virus by immunohistochemistry. In spinal ganglion cells the internal, cytosolic viral proteins (the nucleocapsid, polymerase and matrix proteins) were confined to the perikarya, while the envelope glycoproteins (the haemagglutinin-neuraminidase and fusion proteins) also appeared in the axon-like processes. All five proteins occurred in the dendrite-like processes of spinal cord neurons. In both types of neuron the cytosolic viral proteins showed a pattern of distribution similar to that observed for the microtubule-associated protein MAP2. The segregated occurrence of the viral envelope and cytosolic proteins in axons may prevent virus assembly in axons and limit long-distance spread of paramyxoviruses in the nervous system.

Animals↗
PubMed1990

A subgroup-specific antigenic site in the G protein of respiratory syncytial virus forms a disulfide-bonded loop.

An antigenic site (represented by 15 amino acids, residues 174 to 188, designated peptide 12) of the large glycoprotein G of respiratory syncytial virus was demonstrated to be subgroup specific in peptide enzyme-linked immunosorbent assay tests with murine monoclonal antibodies and human postinfection sera. The role of individual amino acids in this subgroup-specific site was determined by use of single-amino-acid-deletion sets of peptides. When monoclonal antibodies were reacted with the deletion sets, a broad amino acid dependence of 11 or 12 residues, Cys-176 (Ile-175 in subgroup B) to Cys-186, was found. Human postinfection sera exhibited a narrower reaction profile (for subgroup A, Cys-182 to Trp-183; for subgroup B, Cys-176 to Lys-183). Reduction of peptides on microtiter plates by treatment with dithiothreitol completely destroyed their antigenic activity in tests with monoclonal antibodies and human postinfection sera of subgroup B. A variant of peptide 12 containing all four cysteines of the G protein (represented by 16 amino acids, residues 172 to 187, designated peptide 12var) also was subgroup specific. We concluded that the activity of the antigenic site in tests with monoclonal antibodies for subgroups A and B appears to depend on intrapeptide disulfide bonds. Reactions with postinfection sera of subgroup B also may depend on a disulfide bond. In contrast, postinfection sera of subgroup A appeared to have the capacity to identify a subgroup-specific site in a linear form of the selected 15-amino-acid-long peptide. Treatment of peptides with dithiothreitol had no effect on their antigenic activity in tests with human postinfection sera of subgroup A. These findings have relevance for molecular engineering of peptide antigens for use in respiratory syncytial virus subgroup-specific site-directed serology.

Amino Acid Sequence↗
PubMed1989

Comparison of non-ionic detergents for extraction and ion-exchange high-performance liquid chromatography of Sendai virus integral membrane proteins.

The integral membrane proteins of Sendai virus haemagglutinin-neuraminidase (HN) and fusion protein (F) were extracted from purified virions with 2% of a non-ionic detergent, i.e., polyoxyethylene alkyl ethers varying by 8-14 hydrocarbon units in the alkyl chain and by 4-8 ethylene glycol units in the oxyethylene chain. Triton X-100 and octyl glucoside were included as reference detergents. The hydrophile-lipophile balance (HLB) and the critical micelle concentration (CMC) of the detergents were determined. A decrease in length of the oxyethylate by 8-5 ethylene glycol units and an increase in the alkylate by 8-12 hydrocarbon units resulted in higher yields of extracted proteins. The highest yields were obtained for C12E5 with an HLB of 11.7. Yields of extracted protein could be correlated with the HLB values of the polyoxyethylene alkyl ethers. The structural integrity of HN and F was not affected during extraction by either detergent as measured by their reactivity with monoclonal antibodies directed against native HN and F. Extracts were subjected to anion-exchange high-performance liquid chromatography (HPLC) on a Mono Q column in the presence of 0.1% of the detergent used for extraction. Eluate fractions were analysed by sodium dodecyl polyacrylamide gel electrophoresis and recoveries of HN and F protein were determined by size-exclusion HPLC. The immunological activity of HN and F was tested in an enzyme-linked immunosorbent assay. The highest recoveries of HN and F (80%) were obtained with C10E5 in the elution buffer. HN and F were partially purified and the immunological activity was well preserved.

Animals↗
PubMed1989

Comparison of ion-exchange high-performance liquid chromatography columns for purification of Sendai virus integral membrane proteins.

The recovery and separation of the integral membrane proteins, the haemagglutinin-neuraminidase (HN) and the fusion protein (F), from a Sendai virus detergent extract were compared on three different ion-exchange high-performance liquid chromatography (IE-HPLC) columns: Mono Q, TSK DEAE-NPR and Zorbax BioSeries SAX. The detergent, either 1-O-n-octyl-beta-glucopyranoside (octylglucoside) or decyl polyethylene glycol-300 (decyl PEG-300), used for extraction of HN and F proteins from the virions, was also present in the elution buffers at a concentration of 0.1%. Recovery of HN and F proteins was primarily dependent on the detergent present in the eluent, resulting in yields of HN varying from 18 to 28 and 56 to 67%, when octylglucoside and decyl PEG-300, respectively, were used. The highest yield for HN protein was obtained by separation on either a Mono Q or a TSK DEAE-NPR column with decyl PEG-300 as the additive. Yields of F protein were lower, and the highest recovery of 46% was found in the presence of decyl PEG-300 by separation on the Mono Q column. Analysis of the fractions by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and by size-exclusion HPLC indicated that the HN protein eluted in the presence of decyl PEG-300 from the Mono Q and the TSK DEAE-NPR columns was obtained in pure form, while the F protein was slightly contaminated with HN. Analysis of the fractions with monoclonal antibodies directed against conformational epitopes of HN and F proteins indicated that after IE-HPLC the conformation of the proteins is largely retained.

Chromatography, Gel↗
PubMed1989

Naturally occurring canine distemper virus encephalitis: distribution and expression of viral polypeptides in nervous tissues.

Formalin-fixed brain tissues from 16 dogs with naturally occurring canine distemper virus (CDV) infection were investigated immunohistochemically by a panel of eight monoclonal antibodies (mAb) directed against four structural CDV proteins. Three mAb recognizing different epitopes of the polymerase (P-1, P-2, P-3) protein, two clones identifying different epitopes on each, the nucleocapsid (NP-1, NP-2) and fusion (F-2, F-3) protein, and one mAb directed against the hemagglutinin (H-2) protein were used. The immunoreactivity of the clones was tested on formalin-fixed, paraffin-embedded Vero cells, which were lytically infected with the neurotropic R252 (R252-CDV) or the Onderstepoort (CDV/Ond) strain of CDV. Clones directed against the H-2 and F-3 epitope recognized CDV/Ond but not R252-CDV. The remaining six clones showed positive immunoreaction with both CDV strains. In vivo expression and distribution of the individual proteins and their epitopes varied substantially between animals and within lesions from the same animal. The NP-2 epitope showed positive immunostaining in all 16 cases. The P-2 epitope was demonstrated in 13, the NP-1 epitope in 12, the P-3 epitope in 9, and the P-1 epitope in 3 brains, but staining was severely reduced compared with the NP-2 epitope and restricted to areas with strong NP-2 expression. Immunostaining was prominent in early and subacute and reduced in chronic demyelinating lesions. mAb directed against the H and F protein showed no immunoreaction in diseased brains.

Animals↗
PubMed1989

Antigenic characterization of respiratory syncytial virus associated with acute respiratory infections in Uruguayan children from 1985 to 1987.

The occurrence of subgroup A and B strains of respiratory syncytial virus (RSV) was studied during three epidemic years, 1985 to 1987, in Uruguay. A set of monoclonal antibodies was selected according to their reactivity with local RSV isolates and used for the typing of RSV directly in nasopharyngeal cells by indirect immunofluorescence. Of 77 specimens, 69 could be typed as belonging to subgroup A or B, 5 could not be typed with the restricted set of monoclonal antibodies employed, and 3 reacted with both subgroup-specific antibodies. In 1985 and 1986 subgroup A predominated, accounting for 65.7% of all typed specimens, but in 1987 subgroup B surpassed subgroup A, accounting for 82.4% of the samples.

Antibodies, Monoclonal↗
PubMed1988

Effect of detergents on the structure of integral membrane proteins of Sendai virus studied with size-exclusion high-performance liquid chromatography and monoclonal antibodies.

The integral membrane proteins of Sendai virus, the fusion protein F (Mr = 65,000) and the haemagglutinin-neuraminidase protein HN (Mr = 68,000), were used as a model protein mixture. They were subjected to size-exclusion high-performance liquid chromatography on Superose 6HR columns with eluents containing various additives in order to solubilize the proteins. The effect of the additives on the structure of the membrane proteins was investigated with conformation-dependent monoclonal antibodies, either directed against F or HN protein, and by determination of the haemagglutinating capacity of the HN protein. The results show that the structure of the HN protein is more easily disturbed by eluents than that of the F protein. When the elution conditions are mild, e.g., 0.1% octylglucoside, the structure of both proteins is conserved but no separation is obtained. Elution with a buffer containing 0.05% sarkosyl (dodecyl methylglycine sodium salt) did not affect the structure and resulted in pure F protein. Pretreatment of the Amberlite XAD-2-treated Sendai virus envelope extract with 4% sodium dodecyl sulphate (SDS) and elution with 0.1% SDS in 50 mM sodium phosphate (pH 6.5) altered the structure of the HN protein but resulted in purification of the tetramer and the dimer of the HN protein, and the monomer of the F protein.

Antibodies, Monoclonal↗
PubMed1988

Accentuated antibody response to paramyxoviruses in individuals infected with human immunodeficiency virus.

Sera from 31 human immunodeficiency virus (HIV)-infected patients, representing different clinical stages of HIV infection, were assayed for antibodies against measles and mumps viruses by various serological tests and compared to 23 healthy controls. Sera from four patients (two primary, one asymptomatic, and one acquired immunodeficiency syndrome) exhibited a pronounced antibody response to measles as detected by haemagglutination inhibition and radioimmuno-precipitation assay. The RIPA-positive sera showed increased reactivity to all the viral components and in particular to the haemagglutinin (HA) protein of the virus (Fig. 1). Three of these positive patients also showed a similar response to mumps virus. One of the control sera also showed an increase in antibody titre in measles serological tests. The measles antibodies were shown not be anti-HIV antibodies crossreacting with paramyxoviruses. The reactivity to haemagglutinin was still present when using nonglycosylated measles virus antigen grown in the presence of tunicamycin. Whether the accentuated antibody response is due to polyclonal activation mediated by HIV or to reactivation of the viruses remains to be answered.

AIDS-Related Complex↗
PubMed1988

Laboratory markers for over-attenuation of mumps vaccine virus.

The properties of mumps vaccine virus (Leningrad-3 strain) gradually changed upon passaging in quail embryo fibroblasts, the substrate normally used for mumps vaccine production in the USSR. Alterations were extremely noticeable in the over-attenuated (38th passage) virus variant, and involved (a) poor, if any, antibody response in guinea-pigs, (b) turbid plaque formation, (c) lack of expression in cell culture of fusion protein and reduced expression of polymerase protein, and (d) enrichment by abnormally small, fusion-protein-deficient virus particles. Two other laboratory strains exhibited a similar trend to over-attenuation, though after variable passage numbers. Due to a good inter-correlation, every test (namely, inoculation of guinea-pigs, plaque assay, protein analysis, or immune electron microscopy) is indicative of mumps vaccine over-attenuation and hence might be valuable in seed virus quality control.

Animals↗
PubMed1988

Respiratory syncytial virus epidemics: variable dominance of subgroups A and B strains among children, 1981-1986.

We examined the distribution of subgroups A and B strains from respiratory syncytial virus during five epidemic years from 1981 to 1986 in Huntington, West Virginia. Of 235 infants and children with respiratory syncytial virus infection, 211 had virus reisolated from frozen throat swab specimens for subgroup characterization by reactivity with a panel of monoclonal antibodies to the G, F, NP, M, and P proteins by using an enzyme immunoassay. We identified 160 (75.8%) strains as subgroup A and 51 (24.2%) as subgroup B. Strains of both subgroups were isolated in all years. Small, but approximately equal, numbers of subgroup B strains were isolated each epidemic year. By contrast, subgroup A strains occurred at least three times as often in all years except 1984-1985. The very low number of subgroup A strains isolated during the 1984-1985 epidemic gave dominance to subgroup B strains.

Antibodies, Monoclonal↗
PubMed1988

Parainfluenza virus type 2 haemagglutinin-neuraminidase glycoprotein characterized with monoclonal antibodies.

Thirteen monoclonal antibodies (MAbs) were prepared against human parainfluenza virus type 2 (PIV2). These MAbs reacted with the haemagglutinin-neuraminidase glycoprotein with an Mr of 84K. The MAbs defined one antigenic site which could be divided into five epitopes. A correlation between haemagglutination inhibition (HI) and neutralization activity could be seen although one MAb, which recognized a distinct epitope, showed neutralization and no HI activity to PIV2. The reactivity of the MAbs was tested against Sendai virus, parainfluenza virus type 3, simian virus 5 (SV5), mumps virus, Newcastle disease virus, measles virus and canine distemper virus. Only one MAb showed any cross-reaction with a low HI titre to SV5.

Antibodies, Monoclonal↗
PubMed1988

Respiratory syncytial virus: heterogeneity of subgroup B strains.

In order to investigate further possible structural differences among the two subgroups of respiratory syncytial virus (RSV), we analysed the antigenic characteristics and size of structural proteins of 20 subgroup A and 43 subgroup B strains by their reactions with monoclonal antibodies (MAbs) directed against the proteins of RSV using immunofluorescence, ELISA and radioimmunoprecipitation assays. The latter test also enabled determination of the size of different structural components. The 37 MAbs employed were generated by immunization with both subgroup A and B strains. They represented specificities for distinct epitopes on five different structural proteins. The subgroup A strains proved to be relatively uniform. The fusion (F) protein, nucleoprotein (NP) and matrix (M) proteins of all strains tested had the same Mr and all except one strain had a phosphoprotein (P protein) of the same Mr. The F and P proteins were lower in Mr in B strains compared to A strains, which confirmed previous findings. The Mr of the large surface glycoprotein (G protein) of subgroup A strains varied slightly, probably on the basis of differing glycosylation. By contrast, the subgroup B strains exhibited substantial variation in the Mr of the G and also the P proteins and in reactivity with MAbs directed against the G and F proteins. Three size classes of the P protein were identified in B strains: 33K to 34K, 32K to 33K, and 31K to 32K. Twenty-seven subgroup B strains failed to react with four anti-G MAbs representing a single epitope, G2; the remaining 16 strains reacted with these MAbs. We designated these two sets of variants of B strains B1, which lacked the epitope, and B2, which had the epitope. The B1 strains also varied in the size of the G and P proteins. In contrast, all B2 strains had large G proteins and all except two strains had relatively large P proteins (33K to 34K). All subgroup B1 and B2 strains exhibited the same sizes of NP, F and M proteins. We conclude that the subgroup B strains of RSV include two variants, B1 and B2, and that the major difference between them resides in the G and P proteins.

Antibodies, Monoclonal↗