Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1976 Feb;13(2):425–437. doi: 10.1128/iai.13.2.425-437.1976

Comparison of antigens in sonic and pressure cell extracts of Mycobacterium tuberculosis.

B W Janicki, G L Wright Jr, R C Good, S D Chaparas
PMCID: PMC420630  PMID: 816743

Abstract

Comparisons were made of the yield, chemical content, and biological activity of filtrates and extracts obtained by sonic and pressure cell disruption of bacilli from 4- and 8-week-old Proskauer and Beck cultures of the H37Rv strain (TMC no. 102) of Mycobacterium tuberculosis. The culture filtrates were dialyzed, freeze-dried, reconstituted in saline, and sterilized by membrane filtration. The viable bacilli were washed and resuspended in distilled water and subsequently disrupted either by sonication in the cold for 15 or 30 min or by treatment at 20,000 or 40,000 lb/in2 in a pressure cell. The resulting extracts were clarified by centrifugation, concentrated, and sterilized by filtration. All preparations were adjusted to contain 10 mg of solids (dry weight)/ml and were analyzed quantitatively for protein, deoxyribonucleic acid, ribonucleic acid, polysaccharide, and lipid content. Separation patterns obtained by gradient acrylamide gel electrophoresis, as well as by one- and two-dimensional immunoelectrophoresis, provided the basis for qualitative comparisons of the culture filtrates and cell extracts. Three-point dose-response curves also were used to compare the preparations for skin test reactivity in BCG-vaccinated guinea pigs. It was concluded that, although there were no consistent differences in chemical content or biological activity between the preparations, a 15-min sonic treatment appeared to be the most suitable method for preparation of bacillary extracts based on yield of active components and ease of preparation.

Full text

PDF
435

Images in this article

430Image
on p.430
431Image
on p.431
432Image
on p.432
433Image
on p.433
434Image
on p.434
434Image
on p.434
435Image
on p.435

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Affronti L. F., Wright G. L., Jr, Reich M. Characterization and comparison of mycobacterial antigens by discontinuous pore gradient gel electrophoresis. Infect Immun. 1972 Apr;5(4):474–481. doi: 10.1128/iai.5.4.474-481.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beam R. E., Stottmeier K. D., Kubica G. P. Purified protoplasmic peptides of mycobacteria: isolation of species-specific peptides from protoplasm of mycobacteria. J Bacteriol. 1969 Oct;100(1):195–200. doi: 10.1128/jb.100.1.195-200.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CASTELNUOVO G., DUNCAN M. E., BELLEZZA G. MYCOBACTERIAL ANTIGENS: A STUDY OF ANTIGENS IN BACTERIAL EXTRACTS AND IN CULTURE FILTRATES OF DIFFERENT AGES. Tubercle. 1964 Sep;45:246–254. doi: 10.1016/s0041-3879(64)80015-5. [DOI] [PubMed] [Google Scholar]
  4. Counts J. M., Kubica G. P. Investigation of protoplasmic extracts of tubercle bacilli as skin test antigens. Can J Microbiol. 1968 Oct;14(10):1053–1058. doi: 10.1139/m68-177. [DOI] [PubMed] [Google Scholar]
  5. Glenchur H., Burge R. E., Froman S. Tuberculin activity of mycobacterial fractions obtained by chromatography. Appl Microbiol. 1973 Jan;25(1):5–10. doi: 10.1128/am.25.1.5-10.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Haas H., Davidson Y., Sacks T. Taxonomy of mycobacteria studies by polyacrylamide-gel electrophoresis of cell proteins. J Med Microbiol. 1972 Feb;5(1):31–37. doi: 10.1099/00222615-5-1-31. [DOI] [PubMed] [Google Scholar]
  7. Janicki B. W., Chaparas S. D., Daniel T. M., Kubica G. P., Wright G. L., Yee G. S. A reference system for antigens of Mycobacterium tuberculosis. Am Rev Respir Dis. 1971 Oct;104(4):602–604. doi: 10.1164/arrd.1971.104.4.602. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Kwapinski J. B., Alcasid A. Immunological patterns of "typical" and Runyon's group I mycobacteria determined by cytoplasm antigens. Can J Microbiol. 1972 Jun;18(6):731–734. doi: 10.1139/m72-116. [DOI] [PubMed] [Google Scholar]
  10. Kwapinski J. B., Alcasid A., Palser H. Serological relationships of endoplasm antigens of saprophytic mycobacteria. Can J Microbiol. 1970 Sep;16(9):871–876. doi: 10.1139/m70-147. [DOI] [PubMed] [Google Scholar]
  11. Kwapinski J. B., Alcasid A. Serological relationships between cytoplasms of scotochromogenic mycobacteria. Can J Microbiol. 1970 Dec;16(12):1263–1266. doi: 10.1139/m70-211. [DOI] [PubMed] [Google Scholar]
  12. LARSON C. L., RIBI E., WICHT W. C., LIST R. Skin reactions produced in rabbits by cell walls and protoplasm of Mycobacterium tuberculosis and M. butyricum. Am Rev Respir Dis. 1961 Feb;83:184–193. doi: 10.1164/arrd.1961.83.2P1.184. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Larson C. L., Baker M. B., Baker R., Ribi E. Studies of delayed reactions using protoplasm from acid-fast bacilli as provoking antigen. Am Rev Respir Dis. 1966 Aug;94(2):257–259. doi: 10.1164/arrd.1966.94.2.257. [DOI] [PubMed] [Google Scholar]
  15. Larson C. L., Baker R. E., Baker M. B. A comparison of the ability of protoplasm and culture filtrate (PPD) antigens from Mycobacterium tuberculosis, M. kansasii and M. batteyi to elicit delayed reactions in infected guinea pigs. Am Rev Respir Dis. 1968 Dec;98(6):1055–1057. doi: 10.1164/arrd.1968.98.6.1055. [DOI] [PubMed] [Google Scholar]
  16. Larson C., Ribi E., Baker M. B., List R., Baker R., Wicht W. Skin reactions in rabbits sensitized with typical and atypical mycobacteria and tested with homologous and heterologous protoplasm. Am Rev Respir Dis. 1966 Dec;94(6):923–932. doi: 10.1164/arrd.1966.94.6.923. [DOI] [PubMed] [Google Scholar]
  17. Minden P., McClatchy J. K., Farr R. S. Shared antigens between heterologous bacterial species. Infect Immun. 1972 Oct;6(4):574–582. doi: 10.1128/iai.6.4.574-582.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morris D. L. Quantitative Determination of Carbohydrates With Dreywood's Anthrone Reagent. Science. 1948 Mar 5;107(2775):254–255. doi: 10.1126/science.107.2775.254. [DOI] [PubMed] [Google Scholar]
  19. PATTERSON E. K., DACKERMAN M. E. Nucleic acid content in relation to cell size in the mature larval salivary gland of Drosophila melanogaster. Arch Biochem Biophys. 1952 Mar;36(1):97–113. doi: 10.1016/0003-9861(52)90381-0. [DOI] [PubMed] [Google Scholar]
  20. Pickett M. J., Pedersen M. M., Goldstein D. I., Froman S. The antigens of Mycobacterium tuberculosis. Protein fractions from cytoplasm and culture filtrate. Am Rev Respir Dis. 1968 Mar;97(3):415–422. doi: 10.1164/arrd.1968.97.3.415. [DOI] [PubMed] [Google Scholar]
  21. Roberts D. B., Wright G. L., Jr, Affronti L. F., Reich M. Characterization and comparison of mycobacterial antigens by two-dimensional immunoelectrophoresis. Infect Immun. 1972 Oct;6(4):564–573. doi: 10.1128/iai.6.4.564-573.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SNYDER F., STEPHENS N. A simplified spectrophotometric determination of ester groups in lipids. Biochim Biophys Acta. 1959 Jul;34:244–245. doi: 10.1016/0006-3002(59)90255-0. [DOI] [PubMed] [Google Scholar]
  23. Stottmeier K. D., Beam R. E., Kubica G. P. Purified protoplasmic peptides of mycobacteria: in vivo and in vitro comparison of the species specificity of purified protoplasmic peptides and purified protein derivatives of mycobacterial culture filtrates. J Bacteriol. 1969 Oct;100(1):201–208. doi: 10.1128/jb.100.1.201-208.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Turcotte R., Des Ormeaux Y. Influence of the age of mycobacterial cultures on the protein and carbohydrate composition of tuberculins. Can J Microbiol. 1972 May;18(5):637–645. doi: 10.1139/m72-101. [DOI] [PubMed] [Google Scholar]
  25. Wright G. L., Jr, Affronti L. F., Reich M. Characterization and comparison of mycobacterial antigens by two-dimensional polyacrylamide gel electrophoresis. Infect Immun. 1972 Apr;5(4):482–490. doi: 10.1128/iai.5.4.482-490.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wright G. L., Jr, Roberts D. B. Two-dimensional immunoelectrophoresis of mycobacterial antigens. Comparison with a reference system. Am Rev Respir Dis. 1974 Feb;109(2):306–310. doi: 10.1164/arrd.1974.109.2.306. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES