Abstract
By following careful procedures, mycobacterial ribosomal fractions and ribonucleic acid (RNA) prepared by ethyl alcohol precipitation were obtained which have immunogenic activities similar to the viable attenuated H37Ra cells of Mycobacterium tuberculosis from which they were obtained. This comparison was based on the amount of ribonucleic acid (RNA) present. These preparations consisted of approximately 63% RNA and 37% protein; no deoxyribonucleic acid or polysaccharide was detected by chemical tests. A high correlation was found between the immunogenic activity of a preparation and the per cent increase in hyperchromicity at 260 nm of a ribonuclease-hydrolyzed portion. Final concentrations of sodium dodecyl sulfate higher than 0.25% when used for the preparation of the ribosomal fractions and RNA resulted in significantly lower immune responses and greater variation between experiments. This was not related to the amount of protein present. The stability of the ribosomal and RNA preparations was tested under a variety of conditions. The need for a good protective adjuvant again was shown since mouse serum readily hydrolyzed the RNA. Equal immunity was obtained after immunization by the intraperitoneal and subcutaneous routes; however, no immune response was obtained when the intravenous route was used. Preliminary results with RNA prepared with phenol showed that it was more easily degraded during preparation. This resulted in a lower immune response than was obtained with the RNA prepared with ethyl alcohol.
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Selected References
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- Braun W., Firshein W. Biodynamic effects of oligonucleotides. Bacteriol Rev. 1967 Jun;31(2):83–94. doi: 10.1128/br.31.2.83-94.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CRESTFIELD A. M., SMITH K. C., ALLEN F. W. The preparation and characterization of ribonucleic acids from yeast. J Biol Chem. 1955 Sep;216(1):185–193. [PubMed] [Google Scholar]
- GIERER A., SCHRAMM G. Infectivity of ribonucleic acid from tobacco mosaic virus. Nature. 1956 Apr 14;177(4511):702–703. doi: 10.1038/177702a0. [DOI] [PubMed] [Google Scholar]
- KANAI K., YOUMANS G. P., YOUMANS A. S. Allergenicity of intracellular particles, cell walls, and cytoplasmic fluid from Mycobacterium tuberculosis. J Bacteriol. 1960 Nov;80:615–621. doi: 10.1128/jb.80.5.615-621.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Pinchuck P., Fishman M., Adler F. L., Maurer P. H. Antibody formation: initiation in "nonresponder" mice by macrophage synthetic polypeptide RNA. Science. 1968 Apr 12;160(3824):194–195. doi: 10.1126/science.160.3824.194. [DOI] [PubMed] [Google Scholar]
- ROTH J. S., MILSTEIN S. W. Ribonuclease. I. A new assay method with P32 labeled yeast ribonucleic acid. J Biol Chem. 1952 May;196(2):489–498. [PubMed] [Google Scholar]
- SEVER J. L., YOUMANS G. P. The enumeration of nonpathogenic viable tubercle bacilli from the organs of mice. Am Rev Tuberc. 1957 Feb;75(2):280–294. doi: 10.1164/artpd.1957.75.2.280. [DOI] [PubMed] [Google Scholar]
- WEISS D. W. Vaccination against tuberculosis with nonliving vaccines. I. The problem and its historical background. Am Rev Respir Dis. 1959 Oct;80:495–509. doi: 10.1164/arrd.1959.80.4P1.495. [DOI] [PubMed] [Google Scholar]
- WEISS D. W. Vaccination against tuberculosis with nonliving vaccines. I. The problem and its historical background. Am Rev Respir Dis. 1959 Nov;80:676–688. doi: 10.1164/arrd.1959.80.5.676. [DOI] [PubMed] [Google Scholar]
- WEISS D. W. Vaccination against tuberculosis with nonliving vaccines. I. The problem and its historical background. Am Rev Respir Dis. 1959 Sep;80:340–358. doi: 10.1164/arrd.1959.80.3.340. [DOI] [PubMed] [Google Scholar]
- WEISS D. W., WELLS A. Q. Vaccination against tuberculosis with nonliving vaccines. II. Vaccination of guinea pigs with phenol-killed tubercle bacilli. Am Rev Respir Dis. 1960 Apr;81:518–537. doi: 10.1164/arrd.1960.81.4.518. [DOI] [PubMed] [Google Scholar]
- YOUMANS A. S., YOUMANS G. P. EFFECT OF MITOCHONDRIAL STABILIZERS ON THE IMMUNOGENICITY OF THE PARTICULATE FRACTION ISOLATED FROM MYCOBACTERIUM TUBERCULOSIS. J Bacteriol. 1964 Jun;87:1346–1354. doi: 10.1128/jb.87.6.1346-1354.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
- YOUMANS A. S., YOUMANS G. P. FURTHER STUDIES ON A LABILE IMMUNOGENIC PARTICULATE SUBSTANCE ISOLATED FROM MYCOBACTERIUM TUBERCULOSIS. J Bacteriol. 1964 Feb;87:278–285. doi: 10.1128/jb.87.2.278-285.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
- YOUMANS A. S., YOUMANS G. P. IMMUNOGENIC ACTIVITY OF A RIBOSOMAL FRACTION OBTAINED FROM MYCOBACTERIUM TUBERCULOSIS. J Bacteriol. 1965 May;89:1291–1298. doi: 10.1128/jb.89.5.1291-1298.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- YOUMANS A. S., YOUMANS G. P. NATURE OF THE LABILE IMMUNOGENIC SUBSTANCE IN THE PARTICULATE FRACTION ISOLATED FROM MYCOBACTERIUM TUBERCULOSIS. J Bacteriol. 1964 Oct;88:1030–1037. doi: 10.1128/jb.88.4.1030-1037.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
- YOUMANS G. P., MILLMAN I., YOUMANS A. S. The immunizing activity against tuberculous infection in mice of enzymatically active particles isolated from extracts of Mycobacterium tuberculosis. J Bacteriol. 1955 Nov;70(5):557–562. doi: 10.1128/jb.70.5.557-562.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
- YOUMANS G. P., YOUMANS A. S. The measurement of the response of immunized mice to infection with Mycobacterium tuberculosis va. hominis. J Immunol. 1957 May;78(5):318–329. [PubMed] [Google Scholar]
- Youmans A. S., Youmans G. P. Effect of trypsin and ribonuclease on the immunogenic activity of ribosomes and ribonucleic acid isolated from Mycobacterium tuberculosis. J Bacteriol. 1966 Jun;91(6):2146–2154. doi: 10.1128/jb.91.6.2146-2154.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Youmans A. S., Youmans G. P. Preparation and effect of different adjuvants on the immunogenic activity of mycobacterial ribosomal fraction. J Bacteriol. 1967 Oct;94(4):836–843. doi: 10.1128/jb.94.4.836-843.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Youmans A. S., Youmans G. P. Preparation of highly immunogenic ribosomal fractions of Mycobacterium tuberculosis by use of sodium dodecyl sulfate. J Bacteriol. 1966 Jun;91(6):2139–2145. doi: 10.1128/jb.91.6.2139-2145.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Youmans A. S., Youmans G. P. Ribonucleic acid, deoxyribonucleic acid, and protein content of cells of different ages of Mycobacterium tuberculosis and the ralationship to immunogenicity. J Bacteriol. 1968 Feb;95(2):272–279. doi: 10.1128/jb.95.2.272-279.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Youmans G. P., Youmans A. S. Allergenicity of mycobacterial ribosomal and ribonucleic acid preparations in mice and guinea pigs. J Bacteriol. 1969 Jan;97(1):134–139. doi: 10.1128/jb.97.1.134-139.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]