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. 1985 Feb;82(4):1136–1140. doi: 10.1073/pnas.82.4.1136

Identification of individual interferon-producing cells by in situ hybridization.

R Zawatzky, E De Maeyer, J De Maeyer-Guignard
PMCID: PMC397209  PMID: 3856251

Abstract

Individual interferon (IFN)-producing cells were identified by hybridization in situ followed by autoradiography. cDNAs corresponding to murine IFN-alpha and murine IFN-beta labeled by nick-translation to high specific activity (2-4 X 10(8) dpm/micrograms) with alpha-35S-labeled dATP were used as probes for hybridization with IFN mRNA in mouse C-243 cells induced with Newcastle disease virus. Control experiments with non-induced cells or with non-IFN-related labeled DNA monitored the specificity of the autoradiographic signal. Under optimal conditions of IFN induction, between 15% and 40% of the cells gave a hybridization signal with a mixture of IFN-alpha and -beta probes. Differential hybridization with either the IFN-alpha or -beta probe or a mixture of both, at three different time intervals after induction, revealed that only a small fraction of cells had detectable amounts of IFN-alpha mRNA, whereas in the majority of the positive cells IFN-beta mRNA was present.

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Selected References

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

  1. Brahic M., Haase A. T. Detection of viral sequences of low reiteration frequency by in situ hybridization. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6125–6129. doi: 10.1073/pnas.75.12.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. De Maeyer-Guignard J., Tovey M. G., Gresser I., De Maeyer E. Purification of mouse interferon by sequential affinity chromatography on poly(U)--and antibody--agarose columns. Nature. 1978 Feb 16;271(5646):622–625. doi: 10.1038/271622a0. [DOI] [PubMed] [Google Scholar]
  3. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  4. Gall J. G., Pardue M. L. Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A. 1969 Jun;63(2):378–383. doi: 10.1073/pnas.63.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Higashi Y., Sokawa Y., Watanabe Y., Kawade Y., Ohno S., Takaoka C., Taniguchi T. Structure and expression of a cloned cDNA for mouse interferon-beta. J Biol Chem. 1983 Aug 10;258(15):9522–9529. [PubMed] [Google Scholar]
  6. Kelley K. A., Kozak C. A., Dandoy F., Sor F., Skup D., Windass J. D., DeMaeyer-Guignard J., Pitha P. M., DeMaeyer E. Mapping of murine interferon-alpha genes to chromosome 4. Gene. 1983 Dec;26(2-3):181–188. doi: 10.1016/0378-1119(83)90188-9. [DOI] [PubMed] [Google Scholar]
  7. Montagnier L., Collandre H., De Maeyer-Guignard J., De Maeyer E. Two forms of mouse interferon messenger RNA. Biochem Biophys Res Commun. 1974 Aug 5;59(3):1031–1038. doi: 10.1016/s0006-291x(74)80083-5. [DOI] [PubMed] [Google Scholar]
  8. Oie H. K., Gazdar A. F., Buckler C. E., Baron S. High interferon producing line of transformed murine cells. J Gen Virol. 1972 Oct;17(1):107–109. doi: 10.1099/0022-1317-17-1-107. [DOI] [PubMed] [Google Scholar]
  9. Sehgal P. B. The interferon genes. Biochim Biophys Acta. 1982 Sep 30;695(1):17–33. doi: 10.1016/0304-419x(82)90004-x. [DOI] [PubMed] [Google Scholar]
  10. Shaw G. D., Boll W., Taira H., Mantei N., Lengyel P., Weissmann C. Structure and expression of cloned murine IFN-alpha genes. Nucleic Acids Res. 1983 Feb 11;11(3):555–573. doi: 10.1093/nar/11.3.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Shuttleworth J., Morser J., Burke D. C. Expression of interferon-alpha and interferon-beta genes in human lymphoblastoid (Namalwa) cells. Eur J Biochem. 1983 Jun 15;133(2):399–404. doi: 10.1111/j.1432-1033.1983.tb07476.x. [DOI] [PubMed] [Google Scholar]
  12. Weissmann C., Nagata S., Boll W., Fountoulakis M., Fujisawa A., Fujisawa J. I., Haynes J., Henco K., Mantei N., Ragg H. Structure and expression of human IFN-alpha genes. Philos Trans R Soc Lond B Biol Sci. 1982 Sep 24;299(1094):7–28. doi: 10.1098/rstb.1982.0102. [DOI] [PubMed] [Google Scholar]
  13. Yamamoto Y. Antigenicity of mouse interferons: two distinct molecular species common to interferons of various sources. Virology. 1981 Jun;111(2):312–319. doi: 10.1016/0042-6822(81)90335-4. [DOI] [PubMed] [Google Scholar]
  14. Zawatzky R., Kirchner H., DeMaeyer-Guignard J., DeMaeyer E. An X-linked locus influences the amount of circulating interferon induced in the mouse by herpes simplex virus type 1. J Gen Virol. 1982 Dec;63(2):325–332. doi: 10.1099/0022-1317-63-2-325. [DOI] [PubMed] [Google Scholar]

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