Coates, Helen V. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), Ben R. Forsyth, and R. M. Chanock. Biophysical studies of respiratory syncytial virus. I. Density of respiratory syncytial virus and associated complement-fixing antigens in a cesium chloride density gradient. J. Bacteriol. 91:1263-1269. 1966.-Concentrated fluids from respiratory syncytial (RS) virus-infected tissue cultures (HEp-2 and BEK) were subjected to equilibrium sedimentation in cesium chloride. When two antigenically distinct strains of RS virus (Long and 18537) were tested, approximately 90% of the infectious virus was recovered in a sharp, symmetrical peak with a density of 1.22 to 1.24. In a similar study, unconcentrated virus had a density of 1.25 to 1.27. Two immunologically distinguishable complement-fixing antigens (antigens A and B) were detected at densities of 1.28 to 1.32 and 1.23 to 1.37. In addition, the existence of a third antigen (density of 1.22 to 1.30) was suggested. The possible origin of these antigens is discussed relative to the known properties of RS virus and the other myxoviruses.

Antigens↗

PubMed1966

Biophysical studies of respiratory syncytial virus. II. Identification of two soluble complement-fixing antigens of respiratory syncytial virus.

Forsyth, Ben R. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), Helen V. Coates, and Robert M. Chanock. Biophysical studies of respiratory syncytial virus. II. Identification of two soluble complement-fixing antigens of respiratory syncytial virus. J. Bacteriol. 91:1270-1276. 1966.-Column chromatography of concentrated preparations was used to separate two antigenic components from the "soluble complement-fixing (CF) antigen" of respiratory syncytial (RS) virus. These antigens, designated antigen A and antigen B, stimulated homologous but not heterologous CF antibodies. Only antigen A stimulated the production of neutralizing antibody in guinea pigs. There was a dissociation between the CF and neutralizing antibody responses of guinea pigs immunized with antigen A. The two antigens were shown to have different molecular size, density, and sensitivity to lipolytic agents. Both antigens appeared to be components of the RS viral particle. Serological studies demonstrated that both antigens were formed during RS virus infection of children and adults.

Adult↗

PubMed1966

Density of infectious virus and complement-fixing antigens of two rhinovirus strains.

Dans, P. E. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), B. R. Forsyth, and R. M. Chanock. Density of infectious virus and complement-fixing antigens of two rhinovirus strains. J. Bacteriol. 91:1605-1611. 1966.-Two rhinovirus serotypes (echovirus 28 and HGP) and poliovirus type 1 were banded by isopycnic centrifugation in cesium chloride. The rhinovirus virions had a density of 1.41 g/ml, whereas that of poliovirus was 1.34. Since a number of other enteroviruses also have a density of 1.34 g/ml in cesium chloride, a basic difference in density may exist between the rhinovirus and enterovirus subgroups of the picornavirus family. Whether this difference reflects differences in ribonucleic acid content or binding of cesium ions remains to be determined. In tests with echovirus 28 two peaks of CF activity were detected: one in association with the virion (1.41 g/ml), and a larger peak of lower density (1.30 g/ml). With echovirus 28 antiserum, a heterotypically reactive complement-fixing (CF) antigen was detected in the HGP virus suspension at a density less than that of the virion (1.30 g/ml). This antigen corresponded in density to the less dense CF antigen of echovirus 28.

Animals↗

PubMed1966

Isolation and characterization of fractions of Mycoplasma pneumoniae. I. Chemical and chromatographic separation.

Prescott, B. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), O. Sobeslavsky, G. Caldes, and R. M. Chanock. Isolation and characterization of fractions of Mycoplasma pneumoniae. I. Chemical and chromatographic separation. J. Bacteriol. 91:2117-2125. 1966.-Fractionation of Mycoplasma pneumoniae, cultured on a beef heart infusion-horse serum-yeast extract medium, was carried out by chemical and chromatographic procedures. The chemical method yielded eight fractions consisting of lipid, carbohydrates, and proteins. Four protein-rich fractions were isolated by chromatographing a supernatant fluid of sonically treated organisms on Sephadex G-25. The 12 fractions were tested for serological and antigenic activity in vitro and in vivo. The lipid fraction was serologically active and the relative order of activity of the protein fractions appeared to depend on the amount of lipid present in the molecule. The highly serologically active Sephadex G-25 protein fraction 1 prepared chromatographically contained 15% lipid in the molecule, whereas the less serologically active protein fraction 2 prepared by chemical means contained 2% lipid. The acetone-extracted lipid fraction was chromatographed on thin-layer chromatography plates and found to consist of nine fractions. Serological activity was associated with only the first three spots above the origin. Lipid extracted from the protein fractions seemed to be similar to the acetone-extracted lipid from the sediment of the sonically treated organisms.

Amino Acids↗

PubMed1966

Isolation and characterization of fractions of Mycoplasma pneumoniae. II. Antigenicity and immunogenicity.

Sobeslavsky, O. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), B. Prescott, W. D. James, and R. M. Chanock. Isolation and characterization of fractions of Mycoplasma pneumoniae. II. Antigenicity and immunogenicity. J. Bacteriol. 91:2126-2138. 1966.-Chemical and chromatographic fractions of disrupted Mycoplasma pneumoniae organisms were examined for serological and immunogenic activity. Complement-fixing activity was associated with lipid components, whereas precipitin activity was chiefly associated with polysaccharide components. When chemically extracted lipids were separated by thin-layer silica gel chromatography, only three of the nine fractions exhibited complement-fixing activity. Although lipids were highly active serologically, they were only weakly immunogenic. However, lipids combined with protein in lipoprotein complexes were highly immunogenic, stimulating high levels of complement-fixing, indirect-hemagglutinating, and growth-inhibiting antibodies. The specificity of these antibodies was directed chiefly against the serologically active lipid constituents of the organism. It was suggested that these serologically active lipids are present at the sites on the limiting membrane of M. pneumoniae at which antibody acts to inhibit growth of the organism. Only protein fractions adsorbed to tanned erythrocytes. The main function of protein in the indirect-hemagglutination reaction appeared to be that of serving as a carrier for the serologically active lipids.

Animals↗

PubMed1966

Color test for the measurement of antibody to T-strain mycoplasmas.

Purcell, Robert H. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), D. Taylor-Robinson, D. Wong, and R. M. Chanock. Color test for the measurement of antibody to T-strain mycoplasmas. J. Bacteriol. 92:6-12. 1966.-A metabolic inhibition technique for the measurement of antibody to T-strain mycoplasmas was developed, based upon the ability of T-strain mycoplasmas to metabolize urea with the concomitant production of ammonia, and the ability of specific antiserum to inhibit this ammonia production. Phenol red added to the medium served as an indicator of pH change resulting from ammonia production. Specific antiserum to T-strain mycoplasma T-960 was prepared. The T-strain organism was shown to be serologically distinct from the recognized large-colony mycoplasmas. Antibody to mycoplasma strain T-960 in human sera was demonstrated with the metabolic inhibition technique.

Ammonia↗

PubMed1966

Genetic differentiation by nucleic Acid homology I. Relationships among Mycoplasma species of man.

Reich, Paul R. (National Institutes of Health, Bethesda, Md.), Norman L. Somerson, Carol J. Hybner, Robert M. Chanock, and Sherman M. Weissman. Genetic differentiation by nucleic acid homology. I. Relationships among Mycoplasma species of man. J. Bacteriol. 92:302-310. 1966.-Genetic relatedness among human mycoplasmas was evaluated by measuring the amount of nucleic acid hybrid retained on a membrane filter. Hybrids were formed from deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) derived from representative strains of seven serologically distinct human Mycoplasma species. The results indicate that serologically distinct human Mycoplasma species can also be distinguished by the homology techniques. Low-level cross-reactivity was observed among nucleic acids derived from the seven species. Genetic heterogeneity was demonstrated among three strains of M. salivarium and between two strains of M. orale type 2. In contrast, comparison of three strains and three passage levels of M. pneumoniae revealed them to be indistinguishable. M. pneumoniae appears to be the most distinct of all human mycoplasmas, as shown by both homology and the high buoyant density value of its DNA. Nucleic acids from mycoplasmas which had identical buoyant densities were in some cases differentiable. Mycoplasmas with different DNA buoyant densities were invariably distinguishable by the homology technique.

Journal Article↗

PubMed1966

Genetic Differentiation by Nucleic Acid Homology II. Genotypic Variations Within Two Mycoplasma Species.

Somerson, Norman L. (National Institutes of Health, Bethesda, Md.), Paul R. Reich, Barbara E. Walls, Robert M. Chanock, and Sherman M. Weissman. Genetic differentiation by nucleic acid homology. II. Genotypic variations within two Mycoplasma species. J. Bacteriol. 92:311-317. 1966.-A deoxyribonucleic-ribonucleic acid (DNA-RNA) homology technique was used to determine genetic relatedness among the nucleic acids of eight mycoplasmas which were serologically classified as Mycoplasma hominis type 1. The DNA preparations from these organisms were each found to be distinct. No subgrouping of the M. hominis type 1 strains could be demonstrated. In contrast, when the nucleic acids from six serologically related mycoplasmas which were isolated from tissue cultures were studied, the DNA from these species could not be distinguished. The DNA buoyant densities of the tissue culture isolates were similar. These isolates were closely related genetically to a porcine mycoplasma, M. hyorhinis.

Journal Article↗

PubMed1966