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C R Pringle

Publications and source records attributed to C R Pringle.

At least 55 records · Page 3Linked to original sources
PubMed1993

Immunogenicity and pathogenicity of a triple temperature-sensitive modified respiratory syncytial virus in adult volunteers.

The immunogenicity and pathogenicity of a strain of respiratory syncytial (RS) virus modified by sequential induction of three temperature-sensitive (ts) mutations have been evaluated by intranasal administration to 22 adult volunteers. This modified virus, a triple ts mutant designated ts1C, was derived from a double mutant ts1B evaluated in a previous trial. The original isolate (strain RSS-2) and all its derivatives were propagated throughout in human diploid cells in a specially assigned laboratory. The triple mutant ts1C is unable to multiply in MRC-5 cells at 37 degrees C and above. Following nasal administration of ts1C, immune responses were observed in volunteers with low pre-existing neutralizing antibody titres. The ability of mutant ts1C to induce upper respiratory tract disease in adults was greatly diminished in comparison with the non-ts wild-type virus, but not markedly more so than a previously tested double ts mutant (ts1B) which replicates at 37 degrees C. Mutant ts1C, however, may have greater potential as a live vaccine in view of its inherently greater genetic stability.

Adult↗
PubMed1993

Human adenovirus type 5 recombinants expressing simian immunodeficiency virus macaque strain gag antigens.

The p55 gag gene of simian immunodeficiency virus macaque strain (SIVmac) and the core p27 gag component linked to a synthetic AUG codon have been cloned into adenovirus type 5 vectors to generate either viable E3-replacement or defective E1-replacement viruses. The viruses express the expected SIV proteins in both human and, for the non-defective viruses, monkey cells. A considerable proportion of the p55 produced is exported from the infected cell. These viruses should prove useful both in studies of the immune response to SIV and as components of candidate vaccines aimed specifically at provoking cytotoxic T cell responses.

Adenoviruses, Human↗
PubMed1993

Frequent polymerase errors observed in a restricted area of clones derived from the attachment (G) protein gene of respiratory syncytial virus.

Sequence analysis of a large number of clones derived from the carboxy-terminal one-third of the attachment (G) protein gene of subgroup A respiratory syncytial viruses revealed a region very prone to polymerase errors which resulted mainly in frameshifts because of the insertion or deletion of adenosine residues in some but not all runs of such residues. Such mutations were detected in 14% of clones derived from mRNA, 58% of clones derived from genomic-sense RNA, and 50% of clones derived from in vitro-transcribed RNA. This phenomenon appears to be dependent on the template sequence.

Adenosine↗
PubMed1993

Role of SPECT in differentiating malignant from benign lesions in the lower thoracic and lumbar vertebrae.

The authors categorized 125 spinal lesions in cancer patients and 127 lesions in patients with back pain according to their location in the vertebra on single photon emission computed tomographic (SPECT) images. Forty-four lesions were metastases, all in patients with known malignancy. Lesions in the apophyseal joints were all benign. Lesions manifesting as abnormal uptake projecting beyond the vertebral body surface were osteophytes. Thirty-seven percent of the lesions detected in cancer patients were categorized in either of these two benign categories. Lesions showing focal or diffuse uptake in the body were usually benign (96% and 87%, respectively). Lesions showing uptake in the body and pedicle were usually metastases (83%). When abnormal uptake was seen in both the body and posterior elements but with an intervening normal pedicle, benign disease was the most common cause (93%). It was concluded that the location of lesions on tomographic images provides useful information for differentiation between malignant and benign lesions in the vertebrae.

Diagnosis, Differential↗
PubMed1992

Molecular epidemiology of respiratory syncytial virus: rapid identification of subgroup A lineages.

Methods for the rapid analysis of samples of respiratory syncytial (RS) virus are described using the polymerase chain reaction (PCR) followed by restriction mapping. Isolates (either clinical samples or tissue culture grown virus) can readily be divided into subgroups and then further classified into lineages. These methods enable examination of large numbers of isolates by molecular techniques, thereby facilitating research into the molecular epidemiology of the virus.

Base Sequence↗
PubMed1992

Analysis of relatedness of subgroup A respiratory syncytial viruses isolated worldwide.

Respiratory syncytial virus strains (subgroup A) isolated from around the world during the period 1988-1991 were analysed to determine their relatedness. Analysis was by restriction mapping and nucleotide sequencing following amplification of selected regions of the virus genome by polymerase chain reaction (PCR). Twenty-three viruses of subgroup A isolated from cities in temperate regions of the Northern and Southern hemispheres and the tropics during the period 1988-1991 fell into distinct groupings closely related to four of the six lineages defined in analysis of recurrent epidemics within the same city (Birmingham, UK) during the same period. These observations confirm that multiple lineages of RS virus co-circulate locally, and show that very similar viruses are present simultaneously in widely separated countries.

Amino Acid Sequence↗
PubMed1992

Effect of changes in the nucleotide sequence of the P gene of respiratory syncytial virus on the electrophoretic mobility of the P protein.

A consensus sequence for the P protein gene of the RSN-2 strain of respiratory syncytial (RS) virus was obtained by PCR amplification of cDNA obtained by reverse transcription. This established that the extent of sequence variation between two P genes of strains of antigenic subtype B is similar to that among A strains, confirming the conservation of P genes within subtypes and the divergence of the two antigenic subtypes of RS virus. The P protein of RS virus exhibits anomalous electrophoretic mobility with respect to its molecular weight. In vitro transcription and translation of RSN-2 strain cDNA possessing single point mutations revealed that substitutions involving charged amino acids in the carboxy-terminal region had a marked effect on the electrophoretic mobility of the P protein.

Amino Acid Sequence↗
PubMed1992

Assignment of mutant tsN19 (complementation group E) of respiratory syncytial virus to the P protein gene.

The mutation responsible for the temperature-sensitive (ts) phenotype of mutant tsN19 (complementation group E) of respiratory syncytial virus has been located to the P protein gene. Viral protein synthesis was completely restricted at 39 degrees C, and the tsN19 P protein did not react with an anti-P monoclonal antibody (MAb) (3-5) at 33 degrees C. Reversion of temperature sensitivity restored reactivity with MAb 3-5. Nucleotide sequence determination and in vitro expression of cDNA clones of P mRNA derived from wild-type, tsN19 and non-ts revertant-infected cells, revealed that temperature sensitivity and loss of reactivity with MAb 3-5 were consequences of a Gly----Ser amino acid change at position 172. A low M(r) polypeptide, which represented the C-terminal 93 amino acids of the P protein, was produced by internal initiation in the P open reading frame during in vitro translation, and a similar product was detected transiently in vivo.

Animals↗
PubMed1992

Sequence analysis of the 22K, SH and G genes of turkey rhinotracheitis virus and their intergenic regions reveals a gene order different from that of other pneumoviruses.

The nucleotide and deduced amino acid sequences of three genes of turkey rhinotracheitis virus (TRTV) together with the nucleotide sequences of the relevant intergenic regions were determined. The deduced amino acid sequence of one of the genes shows significant identity (42%) to that of the 22K protein of human respiratory syncytial virus (RSV). The TRTV 22K gene, like that of RSV, has a second open reading frame, although the amino acid sequence deduced from this reading frame does not show any similarity to the equivalent predicted RSV protein. The other two genes and their deduced amino acid sequences do not show any sequence similarity to the genes of other pneumoviruses. However, the hydrophobicity profiles of the predicted proteins do show similarities to those of the small hydrophobic (SH) and attachment protein (G) genes of RSV. The TRTV G gene is 1193 nucleotides in length and encodes a protein of 391 amino acids (M(r) 42984), which is rather larger than the RSV G protein (predicted M(r) 36000). The TRTV SH gene is 589 nucleotides in length, encoding a protein of 174 amino acids (M(r) 18797), which is considerably larger than the size of the RSV SH protein (M(r) 7500). The sequences of the intergenic regions derived from clones of polycistronic mRNAs and polymerase chain reaction products obtained with primers from different genes reveal the order on the virus genome to be 3' F-22K-SH-G 5'. This differs from the gene order of paramyxoviruses and morbilliviruses, which lack a 22K gene (and in some cases a SH gene), and the pneumoviruses RSV and pneumonia virus of mice, which have the F and 22K genes located after the G gene.

Amino Acid Sequence↗
PubMed1992

Sequence analysis of the gene encoding the fusion glycoprotein of pneumonia virus of mice suggests possible conserved secondary structure elements in paramyxovirus fusion glycoproteins.

The gene encoding the fusion (F) glycoprotein of pneumonia virus of mice consists of 1657 bases and contains an open reading frame encoding 537 amino acids which is more similar to the F proteins of pneumoviruses than to those of other paramyxoviruses. Computer-assisted sequence analyses can be combined with data on the antigenicity of various F proteins to suggest a possible arrangement of secondary structure elements common to all pneumovirus and paramyxovirus F proteins.

Amino Acid Sequence↗
PubMed1991

The nucleotide sequences of intergenic regions between nine genes of pneumonia virus of mice establish the physical order of these genes in the viral genome.

We have cloned eight intergenic regions from the Pneumovirus pneumonia virus of mice that link the nine small and medium sized genes previously described (Chambers et al., 1990). The nucleotide sequences of the clones confirm the locations of these genes and their mRNA transcripts in the viral genome. The intergenic regions vary in size from 2-56 nucleotides and show only faint homology to each other or to their analogues in respiratory syncytial virus. Sequence alignments suggest that the location of the transcriptional start site for the mRNA encoding the major nucleocapsid protein of pneumonia virus of mice and respiratory syncytial virus may have altered during virus evolution by gain or loss of a transcriptional start signal.

Base Sequence↗
PubMed1991

Defective synthesis of envelope proteins by temperature-sensitive mutants representing complementation groups B and D of respiratory syncytial virus.

The phenotypes of two complementing temperature-sensitive (ts) mutants of respiratory syncytial (RS) virus indicate that the mutational lesions involve the attachment (G) and matrix (M) proteins of the viral envelope. Synthesis of the G protein was affected in cells infected with mutant tsA2 (complementation group B); the p50 precursor of the G protein was synthesized normally, but further maturation to the fully glycosylated form was defective at 39 degrees C. A non-ts alteration in the efficiency of cleavage of the F0 precursor to the F1 and F2 subunits of the fusion protein was also observed in tsA2-infected cells, which is consistent with the aberrant non-syncytial plaque morphology induced by tsA2 in certain cells. In cells infected with mutant tsN1 (complementation group D) the M protein disappeared from the soluble cytoplasmic fraction soon after synthesis at 39 degrees C and had a slightly decreased electrophoretic mobility. The M protein of non-ts revertants was stable at 39 degrees C, which links the defect in M protein stability with the tsN1 phenotype. However, the aberrant mobility phenotype remained, suggesting pseudoreversion. These results assign two of the eight complementation groups of ts mutants of RS virus.

Cell Line↗
PubMed1991

Genes 1 and 2 of pneumonia virus of mice encode proteins which have little homology with the 1C and 1B proteins of human respiratory syncytial virus.

Genes 1 and 2 of pneumonia virus of mice (PVM) consist of 410 and 571 nucleotides and encode proteins of 113 and 156 amino acids respectively. The proteins show no extensive (gene 1 analogous to 1C) or low (gene 2 analogous to 1B) homology to their presumed counterparts in human respiratory syncytial virus (HRSV). The strongest homology is between regions of approximately 35 amino acids located near the carboxy termini of the gene 2 product and the 1B protein with 29% identity, although a lower level of homology can be detected throughout much of these proteins (18% identity overall). These observations contrast with the conservation of 1C and 1B proteins between subgroups of HRSV and with the conservation of nucleocapsid proteins between HRSV and PVM.

Amino Acid Sequence↗
PubMed1991

Respiratory syncytial virus heterogeneity during an epidemic: analysis by limited nucleotide sequencing (SH gene) and restriction mapping (N gene).

The genes encoding the small hydrophobic (SH) proteins of a series of respiratory syncytial (RS) virus strains were amplified using the polymerase chain reaction, cloned and sequenced. Analysis of the SH gene sequences from 12 RS virus strains isolated between 1956 and 1989 confirmed the homogeneity of the two subgroups. A and B, previously defined serologically. Although there is only 76% deduced amino acid sequence identity of SH proteins between subgroups, there was little variation in deduced amino acid sequences within the subgroups; nucleotide homologies within the subgroups ranged between 93% and 99%. Forty-two isolates of RS virus from a single epidemic season (autumn/winter 1989) were also examined to determine their relatedness. For these isolates regions of both the SH and nucleocapsid protein genes of each isolate were amplified and these regions were further analysed by direct nucleotide sequencing or restriction mapping. It was possible to discriminate at least six different lineages (or substrains) of RS virus circulating at the same time and in the same locality.

Amino Acid Sequence↗
PubMed1991

Sequence of the major nucleocapsid protein gene of pneumonia virus of mice: sequence comparisons suggest structural homology between nucleocapsid proteins of pneumoviruses, paramyxoviruses, rhabdoviruses and filoviruses.

The complete nucleotide sequence of gene 3 of pneumonia virus of mice has been determined, and the 5' end of the mRNA mapped using a modification of the polymerase chain reaction technique. The gene contains a single open reading frame, beginning with a 5'-proximal AUG initiation codon, encoding a polypeptide with a predicted Mr of 43141. Expression of the gene 3 protein in Escherichia coli and in vitro showed that it reacted with virus-specific antiserum and comigrated with the major nucleocapsid (N) polypeptide. The predicted amino acid sequence has extensive identity with that of the N protein of human respiratory syncytial virus. Comparisons with the amino acid sequences of N proteins of other paramyxoviruses, vesicular stomatitis virus and Ebola virus suggest that these proteins may have retained much of the same structure. These regions of conserved structure would most likely have the common functions of RNA binding and protein/protein interactions in the virus nucleocapsid.

Amino Acid Sequence↗
PubMed1991

Identification of variable domains of the attachment (G) protein of subgroup A respiratory syncytial viruses.

We have previously classified isolates from a respiratory syncytial (RS) virus epidemic into distinct lineages by restriction mapping and nucleotide sequencing of parts of the nucleocapsid protein and small hydrophobic protein genes, which are areas of the genome not considered to be under immunological pressure. This study has now been extended by the determination of the nucleotide sequences of the attachment (G) protein genes of isolates from each subgroup A lineage. Deduced amino acid identities of the G proteins ranged between 80% and 99%, corresponding closely to the previously determined relatedness of the lineages. The amino acid variability was not evenly distributed; in the extracellular part of the protein there was a sharply defined hypervariable domain which was separated from a more extended variable domain by a highly conserved region. Most nucleotide changes in the variable domains were in the first and second positions of the codon triplets. These results suggest that there may be considerable immunological pressure for change in certain areas of the G protein and this may account for the ability of this virus to reinfect individuals repeatedly. The results presented here reflect the pattern of published data comparing prototype strains of the A and B subgroups.

Amino Acid Sequence↗
PubMed1990

Determinants of susceptibility to challenge and the antibody response of adult volunteers given experimental respiratory syncytial virus vaccines.

The virulence and immunogenicity of a wild-type respiratory syncytial (RS) virus together with four temperature sensitive (ts) mutants derived from this isolate were tested by intranasal inoculation into adult volunteers. Resistance to challenge correlated with neutralizing antibody titres in nasal secretions and to a lesser extent in serum. All ts mutants were reduced in virulence. Mutant ts1B caused mild or asymptomatic infections yet induced antibody responses comparable with wild-type RS virus. Ts1B might be developed further to produce a live-virus vaccine.

Adult↗