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. 1973 Aug;8(2):215–221. doi: 10.1128/iai.8.2.215-221.1973

Preliminary Characterization of “Immunogenic” Ribonucleic Acid Derived from Rat Peritoneal Exudate Cells

Stanley J Archer a,1, Carl J Wust a
PMCID: PMC422835  PMID: 4579900

Abstract

An “immunogenic” ribonucleic acid (Im-RNA) has been extracted from peritoneal exudate (PE) cells of rats that were immunized with sheep erythrocytes (SRBC). Following multiple phenol extractions and deoxyribonuclease treatment, the material obtained from PE cells was eluted from diethylaminoethyl-cellulose at 0.55 M NaCl concentration and partially purified in this procedure by a factor of 7- to 10-fold. After column chromatography, Im-RNA was found to be free of antigen based on results using 51Cr-labeled SRBC or 14C-dinitrophenol coupled to methylated bovine serum albumin as antigens. The Im-RNA showed a biphasic hyperchromicity curve when heated. The first phase, from 30 C to 90 C was gradual, accounting for 15.2% hyperchromicity suggestive of transfer RNA melting. No loss in immunogenic activity was observed when the Im-RNA was heated to 90 C. The second phase, from 90 C to 102 C, accounting for 15.2% further hyperchromicity, had a calculated Tm of 96 C. Heating above 90 C resulted in an irreversible loss of immunogenic activity. These results strongly suggested that the RNA fraction contained a highly ordered secondary structure such as might be found with double-stranded nucleic acid. The nature and function of the Im-RNA is discussed.

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Selected References

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

  1. ASKONAS B. A., RHODES J. M. IMMUNOGENICITY OF ANTIGEN-CONTAINING RIBONUCLEIC ACID PREPARATIONS FROM MACROPHAGES. Nature. 1965 Jan 30;205:470–474. doi: 10.1038/205470a0. [DOI] [PubMed] [Google Scholar]
  2. Bell C., Dray S. Conversion of non-immune spleen cells by ribonucleic acid of lymphoid cells from an immunized rabbit to produce gammaM antibody of foreign light chain allotype. J Immunol. 1969 Dec;103(6):1196–1211. [PubMed] [Google Scholar]
  3. Bishop D. C., Pisciotta A. V., Abramoff P. Synthesis of normal and "immunogenic RNA" in peritoneal macrophage cells. J Immunol. 1967 Oct;99(4):751–759. [PubMed] [Google Scholar]
  4. Cruchaud A., Unanue E. R. Fate and immunogenicity of antigens endocytosed by macrophages: a study using foreign red cells and immunoglobulin G. J Immunol. 1971 Nov;107(5):1329–1340. [PubMed] [Google Scholar]
  5. Duke L. J., Harshman S. In vitro induction of anti-DNP antibody by immunogenic RNA from rabbit liver. Immunochemistry. 1971 May;8(5):431–455. doi: 10.1016/0019-2791(71)90506-4. [DOI] [PubMed] [Google Scholar]
  6. Duke L. J., Miller C., Harshman S. Induction of antibody synthesis by purified immunogenic RNA in vitro. Nat New Biol. 1972 Feb 9;235(58):180–182. doi: 10.1038/newbio235180a0. [DOI] [PubMed] [Google Scholar]
  7. Egan M. L., Smyth R. D., Maurer P. H. Association of polypeptides with ribonucleic acid. I. Effect of charge of the polypeptide and immune status of animals. J Immunol. 1971 Aug;107(2):540–546. [PubMed] [Google Scholar]
  8. FISHMAN M., ADLER F. L. Antibody formation initiated in vitro. II. Antibody synthesis in x-irradiated recipients of diffusion chambers containing nucleic acid derived from macrophages incubated with antigen. J Exp Med. 1963 Apr 1;117:595–602. doi: 10.1084/jem.117.4.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. FISHMAN M. Antibody formation in tissue culture. Nature. 1959 Apr 25;183(4669):1200–1201. doi: 10.1038/1831200a0. [DOI] [PubMed] [Google Scholar]
  10. FISHMAN M. Antibody formation in vitro. J Exp Med. 1961 Dec 1;114:837–856. doi: 10.1084/jem.114.6.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. FISHMAN M., HAMMERSTROM R. A., BOND V. P. In vitro transfer of macrophage RNA to lymph node cells. Nature. 1963 May 11;198:549–551. doi: 10.1038/198549a0. [DOI] [PubMed] [Google Scholar]
  12. Friedman H. P., Stavitsky A. B., Solomon J. M. Induction in vitro of antibodies to phage T2: antigens in the RNA extract employed. Science. 1965 Sep 3;149(3688):1106–1107. doi: 10.1126/science.149.3688.1106. [DOI] [PubMed] [Google Scholar]
  13. Gottlieb A. A. Macrophage ribonucleoprotein: nature of the antigenic fragment. Science. 1969 Aug 8;165(3893):592–594. doi: 10.1126/science.165.3893.592. [DOI] [PubMed] [Google Scholar]
  14. Gottlieb A. A., Schwartz R. H. Review. Antigen--RNA interactions. Cell Immunol. 1972 Oct;5(2):341–362. doi: 10.1016/0008-8749(72)90060-3. [DOI] [PubMed] [Google Scholar]
  15. Harshman S., Duke L. J., Six H. Mechanism of antibody biosynthesis. I. Isolation and physical chemical properties of the retained antigen. Immunochemistry. 1969 Mar;6(2):175–188. doi: 10.1016/0019-2791(69)90154-2. [DOI] [PubMed] [Google Scholar]
  16. Juras D., Abramoff P. In vitro induction of sulfanilic acid-specific antibodies by RNA extracted from rat spleen. J Immunol. 1970 Nov;105(5):1244–1252. [PubMed] [Google Scholar]
  17. Miller J. F., Mitchell G. F. Cell to cell interaction in the immune response. I. Hemolysin-forming cells in neonatally thymectomized mice reconstituted with thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):801–820. doi: 10.1084/jem.128.4.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mitchell G. F., Miller J. F. Cell to cell interaction in the immune response. II. The source of hemolysin-forming cells in irradiated mice given bone marrow and thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):821–837. doi: 10.1084/jem.128.4.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mosier D. E., Cohen E. P. Induction and rapid expression of an immune response in vitro. Nature. 1968 Aug 31;219(5157):969–970. doi: 10.1038/219969a0. [DOI] [PubMed] [Google Scholar]
  20. NISHIMURA S., NOVELLI G. D. Resistance of S-RNA to ribonucleases in the presence of magnesium ion. Biochem Biophys Res Commun. 1963 May 3;11:161–165. doi: 10.1016/0006-291x(63)90327-9. [DOI] [PubMed] [Google Scholar]
  21. PEACOCKE A. R., WALKER I. O. The thermal denaturation of sodium deoxyribonucleate. I. The limits. J Mol Biol. 1962 Nov;5:550–559. doi: 10.1016/s0022-2836(62)80128-4. [DOI] [PubMed] [Google Scholar]
  22. Ray P. K., Simmons R. L. Serological studies of enzyme-treated murine lymphoid cells. Proc Soc Exp Biol Med. 1973 Mar;142(3):846–852. doi: 10.3181/00379727-142-37130. [DOI] [PubMed] [Google Scholar]
  23. Roelants G. E., Goodman J. W. The chemical nature of macrophage RNA-antigen complexes and their relevance to immune induction. J Exp Med. 1969 Sep 1;130(3):557–574. doi: 10.1084/jem.130.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Speirs R. S. Multiple cellular and subcellular responses to antigen. Literature review and hypothesis of immunization. Immunochemistry. 1971 Jul;8(7):665–689. doi: 10.1016/0019-2791(71)90206-0. [DOI] [PubMed] [Google Scholar]
  25. Walker W. S. Macrophage functional heterogeneity in the in vitro induced immune response. Nat New Biol. 1971 Feb 17;229(7):211–212. doi: 10.1038/newbio229211a0. [DOI] [PubMed] [Google Scholar]
  26. Wilson D. B., Wecker E. E. Quantitative studies on the behavior of sensitized lymphoid cells in vitro. 3. Conversion of "normal" lymphoid cells to an immunologically active status with RNA derived from isologous lymphoid tissues of specifically immunized rats. J Immunol. 1966 Oct;97(4):512–516. [PubMed] [Google Scholar]
  27. Yamada H., Yamada A. Antibody formation against 2,4-dinitrophenyl-hapten at the cellular level. Hemolytic plaque-in-gel assay for lymphoid cells producing anti-2,4-dinitrophenyl-hapten antibody. J Immunol. 1969 Aug;103(2):357–363. [PubMed] [Google Scholar]
  28. Yang W. K., Novelli G. D. Isoaccepting +RNA's in mouse plasma cell tumors that synthesize different myeloma protein. Biochem Biophys Res Commun. 1968 May 23;31(4):534–539. doi: 10.1016/0006-291x(68)90510-x. [DOI] [PubMed] [Google Scholar]

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