Skip to main content
NCBI home page
Wiley - PMC COVID-19 Collection logoLink to Wiley - PMC COVID-19 Collection
. 2005 Dec 7;34(1):38–44. doi: 10.1002/jmv.1890340107

Localization of rhinovirus replication in vitro with in situ hybridization

Eurico de Arruda III 2,✉, Theodore E Mifflin 3, Jack M Gwaltney Jr 1, Birgit Winther 3, Frederick G Hayden 3
PMCID: PMC7166441  PMID: 1653307

Abstract

To facilitate understanding of human rhinovirus (HRV) pathogenesis, methods were developed for detection of HRV infection in vitro using in situ hybridization (ISH). HRV‐14 RNA probes and oligonucleotide probes representing conserved sequences in the 5′‐non‐translated region were labeled with 35S and used to detect infected HeLa or WI‐38 strain human embryonic lung cells in cytological preparations. ISH was shown to be specific for detection of HRV on a single‐cell basis. Subsequently, in human nasal polyps infected in vitro, both oligonucleotide‐ and riboprobes produced a strong signal in association with ciliated epithelial cells. In human adenoids infected in vitro, a signal was observed in nonciliated epithelial cells. This study shows that HRV replicates in ciliated cells in the epithelium of human nasal polyps infected in vitro, and the presence of viral RNA in non‐ciliated cells of the human adenoid infected in vitro suggests that other cell types may also support rhinovirus replication.

Keywords: picornavirus, RNA probes, oligonucleotide probes, ciliated epithelial cell

References

  1. Berger SL, Birkenmeier CS (1979): Inhibition of intractable nucleases with ribonucleoside‐vanadyl complexes: Isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry 18: 5143–5149. [DOI] [PubMed] [Google Scholar]
  2. Bruce CB, Al‐Nakib W, Almond JW, Tyrrel DAJ (1989): Use of synthetic oligonucleotide probes to detect rhinovirus RNA. Archives of Virology 105: 179–187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Callahan PL, Mizutani, S , Colonnp RJ (1985): Molecular cloning and complete sequence determination of RNA genome of human rhinovirus type 14. Proceedings of the National Academy of Science USA 82: 732–736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cox KH, DeLeon DV, Angerer LM, Angerer RC (1984): Detection of mRNAs in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Developmental Biology 101: 485–502. [DOI] [PubMed] [Google Scholar]
  5. Duechler M, Skern T, Sommergruber W, Neubauer C, Gruendler P, Fogy I, Blaas D, Kuechler E (1987): Evolutionary relationships within the human rhinovirus genus: Comparison of serotypes 89, 2, and 14. Proceedings of the National Academy of Science USA 84: 2605–2609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gama RE, Horsnell PR, Hughes PJ, North C, Bruce CB, Al‐Nakib W, Stanway G (1989): Amplification of rhinovirus specific nucleic acids from clinical samples using the polymerase chain reaction. Journal of Medical Virology 28: 73–77. [DOI] [PubMed] [Google Scholar]
  7. Greve JM, Davis G, Meyer AM, Forte CP, Yost SC, Marlor CW, Kamarck ME, McClelland A (1989): The major human rhinovirus receptor is ICAM‐1. Cell 56: 839–847. [DOI] [PubMed] [Google Scholar]
  8. Gwaltney JM, Jr. , Colonno RJ, Hamparian VV, Turner RB. Rhinovirus (1989): In Schmidt NJ, Emmons RW. (eds): “Diagnostic Procedures for Viral, Rickettsial, and Chlamydial Infections.” Washington: American Public Health Association, pp 579–614. [Google Scholar]
  9. Hamory BH, Hendley JO, Gwaltney JM, Jr. , (1977): Rhinovirus growth in nasal polyp organ culture. Proceedings of the Society for Experimental Biology and Medicine 155: 577–582. [DOI] [PubMed] [Google Scholar]
  10. Hughes PJ, North C, Jellis CH, Minor PD, Stanway G (1988): The nucleotide sequence of human rhinovirus IB: Molecular relationships within rhinovirus genus. Journal of General Virology 69: 49–58. [DOI] [PubMed] [Google Scholar]
  11. Lee JL, Costlow NA (1987): A molecular titration assay to measure transcript prevalence levels. Methods in Enzymology 152: 633–648. [DOI] [PubMed] [Google Scholar]
  12. Maples JA (1985): A method for the covalent attachment of cells to glass slides for use in immunohistochemical assays. American Journal of Clinical Pathology 83: 356–363. [DOI] [PubMed] [Google Scholar]
  13. Palmenberg AC (1989): Sequence alignments of picornaviral capsid proteins In Semler BL, Ehrenfeld E. (eds): “Molecular aspects of picornavirus infection and detection.” Washington: American Society for Microbiology, pp 211–241. [Google Scholar]
  14. Richtsmeier WJ, Shikhani AH (1987): The physiology and immunology of the pharyngeal lymphoid tissue. Otolaryngologic Clinics of North America 20: 219–228. [PubMed] [Google Scholar]
  15. Rossman MG, Arnold E, Erickson JW, Frankenberger EA, Griffith JP, Hecht HJ, Johnson JE, Kamer G, Luo M, Mosser AG, Rueckert RR, Sherry B, Vriend G (1985): The structure of a human common cold virus (rhinovirus 14) and its functional relations to other picornaviruses. Nature 317: 145–153. [DOI] [PubMed] [Google Scholar]
  16. Sheehan DC, Hrapchak BB (1980): Theory and practice of histotechnology. St. Louis: C.V. Mosby, pp 137–158. [Google Scholar]
  17. Skern T, Sommergruber W, Blaas D, Gruendler P, Fraundorfer F, Pieler C, Fogy I, Kuechler E (1985): Human Rhinovirus 2: Complete nucleotide sequence and proteolytic processing signals in the capsid protein region. Nucleic Acids Research 13: 2111–2126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stanway G, Hughes PJ, Mountford RC, Minor PD, Almond JW (1984): The complete nucleotide sequence of a common cold virus: Human rhinovirus type 14. Nucleic Acids Research 12: 7859–7875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Staunton DE, Merluzzi VJ, Rothlein R, Barton R, Marlin SD, Springer TA (1989): A cell adhesion molecule, ICAM‐1, is the major surface receptor for rhinoviruses. Cell 56: 849–853. [DOI] [PubMed] [Google Scholar]
  20. Tomassini JE, Graham D, DeWitt CM, Lineberger DW, Rodkey JA, Colonno RJ (1989): cDNA cloning reveals that the major group rhinovirus receptor on HeLa cells is intercellular adhesion molecule 1. Proceedings of the National Academy of Science USA 86: 4907–4911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tourtellotte WW, Verity AN, Schmid P, Martinez S, Shapshak P (1987): Covalent binding of formalin fixed paraffin embedded brain tissue section to glass slides suitable for in situ hybridization. Journal of Virological Methods 15: 87–99. [DOI] [PubMed] [Google Scholar]
  22. Turner RB, Hendley JO, Gwaltney JM Jr., (1982): Shedding of ciliated epithelial cells in rhinovirus colds. Journal of Infectious Diseases 145: 849–853. [DOI] [PubMed] [Google Scholar]
  23. Turner RB, Winther B, Hendley JO, Mygind N, Gwaltney JM, Jr. , (1984): Sites of viral recovery and antigen detection in epithelial cells during experimental rhinovirus infection. Acta Oto‐Laryn‐gologica (Stockh) 413 (suppl): 9–14. [DOI] [PubMed] [Google Scholar]
  24. Winther B, Farr B, Turner RB, Hendley JO, Gwaltney JM, Jr. , Mygind N (1984): Histopathologic examination and enumeration of polymorphonuclear leukocytes in the nasal mucosa during experimental rhinovirus colds. Acta Oto‐Laryngologica (Stockh) 413 (suppl): 19–24. [DOI] [PubMed] [Google Scholar]
  25. Winther B, Gwaltney JM, Jr. , Mygind N, Turner RB, Hendley JO (1986): Sites of rhinovirus recovery after point inoculation of upper airway. Journal of the American Medical Association 256: 1763–1767. [PubMed] [Google Scholar]
  26. Winther B, Gwaltney JM Jr., Hendley JO (1990): Respiratory virus infection of mpnolayer cultures of human nasal epithelial cells. American Review of Respiratory Diseases 141: 839–845. [DOI] [PubMed] [Google Scholar]
  27. Wolf JL, Rubin DH, Finberg R, Kauffman RS, Sharpe AH, Trier JS, Fields BN (1981): Intestinal M cells: A pathway for entry of reovirus into the host. Science 212: 471–472. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Medical Virology are provided here courtesy of Wiley

RESOURCES