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THE PREPARATION OF HIGHLY PURIFIED PR8 INFLUENZA VIRUS FROM INFECTED MOUSE LUNGS
1Department of Animal and Plant Pathology of The Rockefeller Institute for Medical Research, Princeton, New Jersey.
Abstract:
Highly purified preparations of PR8 influenza virus were obtained from perfused, infected mouse lungs by a combination of methods involving adsorption of the virus on and elution from chicken red cells and differential centrifugation. Such preparations were found to possess 50 per cent infectivity end-points at 10(-11) to 10(-11.8), and 10(-13) to 10(-14.3) gm. of nitrogen in mice and in chick embryos, respectively. A sedimentation constant of 683 S was obtained for the material and electron micrographs revealed essentially spherical particles about 100 mmicro in diameter. The material was isoelectric at pH 5.4 and chemical analyses on several of the preparations indicated the presence of 10.1 per cent of nitrogen, 1.06 per cent of phosphorus, 6.2 per cent of carbohydrate and about 33 per cent of lipid. Positive tests were obtained for both ribonucleic and desoxyribonucleic acids. The virus was precipitated strongly by antisenim to purified PR8 virus obtained from the allantoic fluid of infected chick embryos and this serum inhibited the agglutination of red cells by the mouse virus to a dilution of about 40,000. In general, the properties of the virus isolated from infected mouse lungs were found to coincide with those of the virus obtained from the allantoic fluid of infected chick embryos.
Insights
Highly purified PR8 influenza virus from mouse lungs exhibits properties similar to virus from chick embryos. This research details the virus
Area of Science:
- Virology
- Microbiology
- Biochemistry
Background:
- Influenza A virus (IAV) poses a significant public health threat.
- Understanding the physical and chemical properties of IAV is crucial for developing effective countermeasures.
- Previous studies have characterized IAV strains isolated from various sources, but detailed analysis of mouse lung-derived PR8 strain is essential.
Purpose of the Study:
- To highly purify and characterize the Puerto Rico 8 (PR8) influenza virus strain isolated from infected mouse lungs.
- To compare the properties of mouse lung-derived PR8 influenza virus with those obtained from chick embryos.
- To provide detailed biochemical and physical data for the PR8 influenza virus.
Main Methods:
- Purification of PR8 influenza virus from infected mouse lungs using adsorption-elution with chicken red blood cells and differential centrifugation.
- Determination of 50% infectivity endpoints in mice and chick embryos.
- Measurement of sedimentation constant (Svedberg units).
- Electron microscopy for particle size and morphology assessment.
- Isoelectric point determination.
- Chemical analysis for nitrogen, phosphorus, carbohydrate, and lipid content.
- Detection of nucleic acids (ribonucleic and desoxyribonucleic acids).
- Serological testing using anti-PR8 serum and red blood cell agglutination inhibition assays.
Main Results:
- Highly purified PR8 influenza virus preparations were obtained from mouse lungs.
- Infectivity endpoints were established at 10(-11) to 10(-11.8) gm nitrogen in mice and 10(-13) to 10(-14.3) gm nitrogen in chick embryos.
- A sedimentation constant of 683 S was determined.
- Electron microscopy revealed spherical particles approximately 100 mmicro in diameter.
- The isoelectric point was found to be pH 5.4.
- Chemical analysis indicated 10.1% nitrogen, 1.06% phosphorus, 6.2% carbohydrate, and 33% lipid.
- Both ribonucleic acid (RNA) and desoxyribonucleic acid (DNA) were detected.
- The virus precipitated strongly with anti-PR8 serum, and this serum inhibited hemagglutination up to a dilution of 40,000.
Conclusions:
- The properties of PR8 influenza virus isolated from mouse lungs are consistent with those of the virus derived from chick embryos.
- The purification and characterization methods employed provide a robust dataset for understanding PR8 influenza virus.
- This study confirms the biochemical composition and physical characteristics of the PR8 strain, aiding in future influenza research and vaccine development.
