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Biomedical subjects

M R Macnaughton

Publications and source records attributed to M R Macnaughton.

At least 19 recordsLinked to original sources

Diagnosis of human coronavirus infections in children using enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay (ELISA) was developed for diagnosing human coronavirus (HCV) infections in children. One hundred and seventy seven nose swabs, throat swabs, and nasopharyngeal aspirates were collected from 30 children suffering from acute respiratory infections. These samples were tested for HCV antigens by ELISA and 28.2% of the samples were shown to be HCV positive. These results indicate that our ELISA should prove useful in the diagnosis of HCV infections in children. Further studies are in progress to extend the ELISA to detect HCVs in experimentally and naturally acquired infections in adults.

Antigens, Viral

Coronaviridae.

The family Coronaviridae comprises a monogeneric group of 11 viruses which infect vertebrates. The main characteristics of the member viruses are: (i) Morphological: Enveloped pleomorphic particles typically 100 nm in diameter (range 60-220 nm), bearing about 20 nm long club-shaped surface projections. (ii) Structural: A single-stranded infectious molecule of genomic RNA of about (5-7) X 10(6) molecular weight. A phosphorylated nucleocapsid protein [mol. wt. (50-60) X 10(3)] complexed with the genome as a helical ribonucleoprotein; a surface (peplomer) protein, associated with one or two glycosylated polypeptides [mol. wt. (90-180) X 10(3)]; a transmembrane (matrix) protein, associated with one polypeptide which may be glycosylated to different degrees [mol. wt. (20-35) X 10(3)]. (iii) Replicative: Production in infected cells of multiple 3' coterminal subgenomic mRNAs extending for different lengths in the 5' direction. Virions bud intracytoplasmically. (iv) Antigenic: 3 major antigens, each corresponding to one class of virion protein. (v) Biological: Predominantly restricted to infection of natural vertebrate hosts by horizontal transmission via the fecal/oral route. Responsible main for respiratory and gastrointestinal disorders.

Antigens, Viral

Prevalence of human coronavirus antibody in the population of southern Iraq.

Sera from adults in Southern Iraq were collected during winter and screened by an enzyme-linked immunosorbent assay for the presence of antibodies to the two antigenic groups of human coronaviruses, the 229E and the OC43 groups: 91% of the sera had antibodies to at least one of the groups, whereas 4 and 5% of the sera had antibodies to only the 229E or OC43 groups, respectively. There was significant correlation between the levels of antibody to the 229E and OC43 group coronaviruses in these sera.

Adult

Infection of a calf with the enteric coronavirus strain Paris.

A tissue-culture-grown enteric coronavirus infected the whole of the gastrointestinal tract and caused enteritis and diarrhoea in a 10-day-old gnotobiotic Friesian calf. Diarrhoea occurred 2 days after inoculation and excretion of virus in faeces and rectal swabs increased until necropsy at 3 days. Virus growth, as detected by indirect immunofluorescence, virus isolation, and thin section electron microscopy, was most extensive in the epithelium of the colon and rectum at 3 days after inoculation. No virus was detected in other organs or in the pleural, peritoneal and oropharyngeal cavities. The origin and host specificity of this coronavirus is discussed.

Animals

Serological relationships of the subcomponents of human coronavirus strain 229E and mouse hepatitis virus strain 3.

Antibodies were raised in rabbits against the structural components of human coronavirus strain 229E and mouse hepatitis virus strain 3, prepared from disrupted virus particles. Hyperimmune sera to the subcomponents showed cross-reactions by enzyme-linked immunosorbent assay between ribonucleoprotein antigens of these viruses, indicating the presence of a common antigen(s). None of the other virus structural components showed any cross-reactivity.

Antigens, Viral

Replication of human respiratory coronavirus strain 229E in human macrophages.

Evidence for the replication of human coronavirus strain 229E (HCV 229E) in macrophages is presented. Virus antigen was detected in macrophages by an immunofluorescent technique 24 h after infection and virus particles were observed in the cisternae of the endoplasmic reticulum by electron microscopy. Giant cells were observed by light and scanning electron microscopy, and large multinucleate cells were seen by thin-section electron microscopy, suggesting that HCV 229E can induce syncytial formation in cultured human macrophages. Furthermore, the production of infectious virus by macrophages was demonstrated by an infectious centre assay.

Antigens, Viral

Occurrence and frequency of coronavirus infections in humans as determined by enzyme-linked immunosorbent assay.

The occurrence of human coronavirus (HCV) infections was analyzed by using sequential sera taken between 1976 and 1981 from adults working in the London area. Antibody rises to HCV 229E and HCV OC43 group viruses were measured in serum samples from these subjects by enzyme-linked immunosorbent assay. HCV infections were found throughout the year, although most occurred during two periods, from June through September and from December through February. There were no marked seasonal differences in either the range of antibody rises obtained or in the HCV groups to which these antibody rises were directed. However, there were more HCV antibody rises during the summer than in the winter. The antibody duration varied considerably, but had a mean of 3.5 months. Finally, the frequency of HCV infection per person was calculated to be 1 per 7.8 months.

Adult

Antigenicity of mouse hepatitis virus strain 3 subcomponents in C57 strain mice.

C57 strain mice were inoculated intraperitoneally with denatured mouse hepatitis virus strain 3 particles and virus surface projection, membrane and ribonucleoprotein subcomponents, obtained from detergent treated purified virus preparations. All immunised animals developed high levels of serum antibody directed against the respective antigens, detectable by enzyme-linked immunosorbent assay. Mice that had been immunised with denatured virus particles or surface projections were protected against infection with mouse hepatitis virus strain 3, whereas immunisation with virus membrane or ribonucleoprotein subcomponents failed to protect mice against virus challenge.

Animals

Ribonucleoprotein of avian infectious bronchitis virus.

The ribonucleoprotein (RNP) of avian infectious bronchitis virus (IBV) was examined by electron microscopy after shadowing with carbon/platinum. Linear RNP strands up to 6.7 microns in length, from three IVB strains, were sensitive to both pancreatic RNase and to proteases. These strands were obtained from spontaneously disrupted complete particles but not from disrupted incomplete particles that lacked RNP. They were also released from Nonidet P40-disrupted particles and could be isolated on sucrose density gradients at a density of 1.27 g/ml. In some cases, helical RNP complexes associated with virus particles were observed that were similar to RNPs of human coronavirus strain 229E and mouse hepatitis virus strain 3.

Centrifugation, Density Gradient

The distribution of human coronavirus strain 229E on the surface of human diploid cells.

The distribution of human coronavirus strain 229E (HCV 229E) particles on the surface of human diploid (MRCc) cells was examined. Virus particles showed a totally random distribution on fixed cells and on cells to which virus had been adsorbed in the cold. A marked redistribution of virus particles was observed on warming virus-cell preparations to 33 degrees C for 20 min, the peripheral areas of the cell becoming relatively devoid of virus particles while the majority of particles were now located some distance from the edge of the cell. Redistribution did not occur in the presence of metabolic inhibitors.

Azides

Antibody to virus components in volunteers experimentally infected with human coronavirus 229E group viruses.

Antibody rises to various virus subcomponents were measured by enzyme-linked immunosorbent assay in the paired sera of volunteers experimentally infected with human coronavirus 229E group viruses. Most of the antibody made during infection was directed against the virus surface projections, with only small amounts of antibody made against membrane or ribonucleoprotein components.

Antibodies, Viral

Two antigenic groups of human coronaviruses detected by using enzyme-linked immunosorbent assay.

Paired sera from volunteers inoculated with one of the five recently isolated strains of human coronavirus (HCV), AD, GI, HO, PA, and RO, none of which has been grown in tissue culture, or with strain OC38 were tested against coronavirus antigens by enzyme-linked immunosorbent assay. When HCV strains OC43, 229E, or the 229E-related tissue culture-adapted strains PR and TO were used as antigens, it was shown that all strains fell into one of two antigenic groups. The HCV OC43 group was comprised of strains OC43, GI, HO, and RO, and the HCV 229E group contained strains AD and PA as well as the tissue culture-adapted strains PR, TO, and KI. Enzyme-linked immunosorbent assay of the paired sera with the coronavirus mouse hepatitis virus strain 3 as antigen confirmed the relationship of this virus to the HCV OC43 group but not to the HCV 229E group.

Adult

The polypeptides of human and mouse coronaviruses. Brief report.

The polypeptide compositions of two coronaviruses, human coronavirus strain 229E (HCV229E) and mouse hepatitis virus strain 3 (MHV3), were characterised on polyacrylamide gels. Similar polypeptide patterns were observed for both viruses consisting of large surface projection glycopolypeptides of mol. wt. 160,000 and 105,000 for HCV229E, and 170,000 and 90,000 for MHV3, two small polypeptides of mol. wt. varying from 24,000 to 20,000, and a polypeptide of mol. wt. 50,000. The results are discussed with respect to previous reports of the polypeptides of these and other coronaviruses.

Coronaviridae

Two particle types of avian infectious bronchitis virus.

Two distinct types of avian infectious bronchitis virus (IBV) particles were isolated on sucrose density gradients. The higher density particles banded at 1.18 g/ml, had typical coronavirus morphology and contained all the structural polypeptides and a complete genome. The less dense particles of density 1.13 g/ml appeared to have typical coronavirus morphology, although they were much more flattened than the more dense particles. Furthermore, these particles lacked the ribonucleoprotein polypeptide and the genome, although all the other polypeptides were present in the same amounts as in the denser particles.

Centrifugation, Density Gradient