Purification and Character of the Swine Influenza Virus

Taylor, A. R.; Sharp, D. G.; McLean, Ian W.; Beard, Dorothy; Beard, J. W.; Dingle, John H.; Feller, Anita

doi:10.1126/science.98.2557.587

Cited by 26 publications

(34 citation statements)

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“…Preparation of Virus for Assay.--Higldy purified preparations of the PR8 and Lee strains of influenza virus were obtained from allantoie fluids of infected chick embryos by a combination of the methods of differential centrifugation and adsorption on and elution from chicken red cells (8)(9)(10)(11)(12). Such preparations were found to consist of particles which were highly active biologically and which were uniform in size, in electrochemical behavior, and in serological reactions (11).…”

Section: Methods and Findingsmentioning

confidence: 99%

“…In view of these findings, it was naturally of interest to determine whether or not similar chemical differences exist between strains or types of an animal virus. Highly purified preparations of influenza viruses (8)(9)(10)(11)(12) were available for this purpose and the PR8 strain of influenza A and the Lee strain of influenza B were chosen for comparison. These viruses produce clinically indistinguishable diseases (13) and appear t 9 be very similar in gross chemical properties (9).…”

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confidence: 99%

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Amino Acid Composition of Highly Purified Viral Particles of Influenza a and B

Knight¹

1947

The Journal of Experimental Medicine

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Microbiological assays for amino acids were made on hydrolysates of four to five highly purified preparations each of influenza A virus (PR8 strain) and influenza B virus (Lee strain). The results of the assays indicated that these strains of influenza virus contain approximately the same amounts of alanine, aspartic acid, glycine, histidine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, and valine. However, significant differences were found in the values for arginine, glutamic acid, lysine, tryptophane, and tyrosine. It is believed that these differences may provide, at least in part, a chemical explanation for some of the differing properties of the PR8 and Lee strains of influenza viruses.

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Section: Methods and Findingsmentioning

confidence: 99%

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confidence: 99%

Amino Acid Composition of Highly Purified Viral Particles of Influenza a and B

Knight¹

1947

The Journal of Experimental Medicine

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“…It has been shown that high speed eentrlfugation of infectious allantoic fluid for 20 to 30 minutes will sediment the PR8 virus activity almost quantitatively (6,10,11). Therefore, it was considered of interest to determine how much normal protein would sediment in 30 minutes as compared with the yield obtained in the usual 90 minute period.…”

Section: Relation Of Time Of Centrifugation To Yield Of Normal Proteinmentioning

confidence: 99%

A Sedimentable Component of Allantoic Fluid and Its Relationship to Influenza Viruses

Knight¹

1944

Journal of Experimental Medicine

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Macromolecular material was isolated from normal allantoic fluid by a centrifugation procedure comparable to that currently employed for the concentration and purification of influenza viruses. The yield of material was found to vary with the age of the embryo, reaching a maximum average value after 14 days of incubation at 39°C. of about 0.02 mg. per ml. of allantoic fluid. The purified material was found to contain protein, carbohydrate, and lipid and to have a general composition similar to purified preparations of PR8 influenza virus. A typical preparation of normal material had an isoelectric point at pH 2.3. Sedimentation studies indicated that the normal material can give a variety of sedimentation constants depending upon the concentration and viscosity of the preparations. The sedimentation constant, corrected for viscosity, of the major component of a fresh preparation was 170 S. The diameters of the predominant particles shown in electron micrographs of the normal material and of preparations of PR8 influenza virus were about 40 and 100 mµ, respectively. Serological tests indicated that the normal material is a good antigen and that preparations of both A and B types of influenza virus obtained from allantoic fluids by centrifugation show a strong serological relationship to the normal material. Freezing and thawing of allantoic fluid, and repeated adsorption of virus on red cells, failed to provide a practical basis for the separation of normal protein from the virus entity in the case of PR8 virus. In the cases of similar preparations of F12 and of Lee viruses, a partial separation of a small component was accomplished by fractional centrifugation and this component and the normal protein were shown to be identical or very closely related. Antiserum to the purified normal material inhibited red cell agglutination by A and B types of influenza virus at serum dilutions of 600 to 700, but failed to show significant neutralizing capacity in chick embryo and in mouse tests at a serum dilution of 100. Rabbit antiserum to purified preparations of PR8 virus gave a 50 per cent red cell agglutination inhibition endpoint at a serum dilution of 112,000. Some of the implications of the findings are discussed.

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“…Although these authors state that their conclusions are wholly dissimilar to those of Chambers and associates, it may be observed that the experimental centrifuge data obtained by the two groups of workers are nevertheless quite similar in certain respects. For example, the experiment of Taylor, Sharp, Beard, Beard, Dingle, and Feller (10), in which the rate of sedimentation of influenza virus chick red cell agglutinating activity was compared with the rate of sedimentation of the Shope papilloma virus, indicates that about 5 per cent of the influenza virus sedimented at a rate less than that of the papilloma virus. This result could be regarded as evidence that about 5 per cent of the influenza virus is smaller than the papilloma virus which has a particle diameter of about 40 mg. A similar interpretation could be offered for the results obtained in the centrifugation of PR8 allantoic fluid for 1 hour at 27,000 g, following which about 25 per cent of the virus activity was found in the supernatant liquid.…”

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confidence: 99%

“…I n addition, it appeared desirable to investigate the components present in different virus strains because of the ease with which a component having a sedimentation constant of about 30 S was demonstrated to exist in F 12 aUantoic fluid (8) and the observation that no similar component could be found in PR8 virus preparations (10). The question as to whether preparations of virus having particle diameters of 80 to 120 mt~ represent aggregates of smaller infectious units also required elucidation.…”

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The Size of Influenza Virus

Stanley¹

1944

Journal of Experimental Medicine

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The sedimentation behavior of influenza virus in dilute solutions of electrolyte was found to be quite variable. At times the virus activity appeared to sediment at a rate comparable with that of particles about 80 to 120 mµ in diameter, at other times at a rate comparable with that of particles about 10 mµ in diameter, and at still other times the bulk of the activity appeared to sediment at a rate comparable with that of the larger particles and the residual activity at a rate comparable with that of the smaller particles. However, in the presence of a sucrose density gradient, the virus activity was always found to sediment with a rate comparable to that of particles about 80 to 120 mµ in diameter; hence it appeared that the variable sedimentation behavior in dilute electrolyte solution was due to convection or mechanical disturbances during centrifugation. About 30 per cent of the high molecular weight protein present in the allantoic fluid of chick embryos infected with the F 12 strain of influenza virus was found to consist of a component having a sedimentation constant of about 30 S, and hence a probable particle diameter of about 10 mµ. The residual protein of high molecular weight was present in the form of a component having a sedimentation constant of about 600 S, and hence a probable particle diameter of about 70 mµ. The proportion of the 30 S component in allantoic fluid of chick embryos infected with the PR8 strain of influenza virus was found to be considerably less. The 600 S and 30 S components of F 12 allantoic fluid were purified and separated by differential centrifugation. The purified preparations of the 600 S component were found to possess a specific virus activity from 100 to over 10,000 times that of the purified preparations of the 30 S component, the difference in activity apparently depending only on the degree of fractionation of the two components. The purified 30 S component was found to sediment normally in the presence of 12 per cent sucrose, whereas the small residual virus activity of such preparations was found to sediment in the presence of a sucrose density gradient with a rate comparable to that of much heavier particles. It is concluded that influenza virus activity is not associated with material having a particle diameter of about 10 mµ, but is associated solely with material having a sedimentation constant of about 600 S and hence a probable particle diameter of about 70 mµ.

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Purification and Character of the Swine Influenza Virus

Cited by 26 publications

References 2 publications

Amino Acid Composition of Highly Purified Viral Particles of Influenza a and B

Amino Acid Composition of Highly Purified Viral Particles of Influenza a and B

A Sedimentable Component of Allantoic Fluid and Its Relationship to Influenza Viruses

The Size of Influenza Virus

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