Interference with Cell Viability and Poliovirus Multiplication by Polyinosinic-Polycytidylic Acid Free

Abstract

SUMMARY

The double-stranded complex of polyinosinic and polycytidylic acids (poly I:C) inhibits the replication of poliovirus in Detroit cells. By a plaque reduction assay a linear relationship was found between the logarithm of the plaque reduction % and the logarithm of the poly I:C concentration, with about 50% inhibition by 0.1 g./ml. of poly I:C.

The maximum hypochromic effect at 233 nm. on complexing of the polynucleotides occurred with 6 volumes of poly I and 4 volumes of poly C from equimolar solutions of the two homopolynucleotides. Gel-filtration of such a complex on Sepharose 4 B separated double and single strands. Of the added poly I 7% remained as single strands at the conditions of maximum hypochromic effect.

The poly C was found to have a chain length several times larger than that of poly I.

Cell multiplication proceeded at normal rate for 48 hr in a spinner culture with 5 g./ml. of poly I:C. With 25 g./ml. no cell multiplication took place within 3 days, and 65% of the cells were dead, whereas in a control culture the cell density doubled and only 15% of cells were dead.

Isolation of RNA from the cells 3 hr after addition of the inducer showed about 20% higher incorporation of [C]uridine-2 into the treated cells. The increase of incorporation was evenly distributed in all RNA bands except for the 32 RNA band which contained about 60% more C than the 32 RNA of the control.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-9-2-111
1970-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/9/2/JV0090020111.html?itemId=/content/journal/jgv/10.1099/0022-1317-9-2-111&mimeType=html&fmt=ahah

References

  1. Bausek H. G., Merigan T. C. 1969; Cell interaction with a synthetic polynucleotide and interferon production in vitro. Virology 39:491
    [Google Scholar]
  2. De Clercq E., Merigan T. C. 1969; Requirements for a stable secondary structure for the antiviral activity of polynucleotides. Nature, London 222:1148
    [Google Scholar]
  3. Eagle E. 1959; Amino acid metabolism in mammalian cell cultures. Science, New York 130:432
    [Google Scholar]
  4. Field A. K., Tytell A. A., Lampson G. P., Fulleman M. R. 1967; Inducers of interferon and host resistance. II. Multistranded synthetic polynucleotide complexes. Proceedings of the National Academy of Sciences of the United States of America 58:1004
    [Google Scholar]
  5. Kawade Y., Ujihara M. 1969; Non-inducing RNA antagonizes the induction of interference with animal virus infection. Nature, London 221:569
    [Google Scholar]
  6. Levy H. B., Law L. W., Rabson A. S. 1969; Inhibition of tumor growth by polyinosinic-polycytidylic acid. Proceedings of the National Academy of Sciences of the United States of America 62:357
    [Google Scholar]
  7. Loening U. E. 1967; The fractionation of high-molecular-weight ribonucleic acids by polyacrylamide gel electrophoresis. Biochemical Journal 102:251
    [Google Scholar]
  8. Margolis S. A., Levy H. B. 1969; The action of poly I:C on the RNA metabolism of cultured mouseL cells. Federation Proceedings. Federation of American Societies for Experimental Biology 28:816
    [Google Scholar]
  9. Michelson A. M., Massoulie J., Guschlbauer W. 1967; Progress in Nucleic Acid Research and Molecular Biology. New York: Academic Press
    [Google Scholar]
  10. Penman S. 1966; RNA metabolism in HeLa cell nucleus. Journal of Molecular Biology 17:117
    [Google Scholar]
  11. Scherrer K., Darnell J. E. 1962; Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochemical and Biophysical Research Communications 7:486
    [Google Scholar]
  12. Vilcek J., Ng MUN H., Friedman-Kien A. E., Krawciw T. 1968; Introduction of interferon synthesis by synthetic double-stranded polynucleotides. Journal of Virology 2:648
    [Google Scholar]
  13. Wagner E. K., Katz L., Penman S. 1967; The possibility of aggregation of ribosomal RNA during hot phenol-SDS deproteinization. Biochemical and Biophysical Research Communications 28:152
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-9-2-111
Loading
/content/journal/jgv/10.1099/0022-1317-9-2-111
Loading

Data & Media loading...

Most cited Most Cited RSS feed