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The Use of Primary Human Cells (Fibroblasts, Monocytes, and Others) to Assess Human Cytomegalovirus Function

Part of the Methods in Molecular Biology book series (MIMB,volume 1119)

Abstract

The extensive tropism of human cytomegalovirus (HCMV) results in the productive infection of multiple cell types within the human host. However, infection of other cell types, such as undifferentiated cells of the myeloid lineage, gives rise to nonpermissive infections. This has been used experimentally to model latent infection which is known to be established in the pluripotent CD34+ hematopoietic progenitor cell population resident in the bone marrow in vivo. The absence of a tractable animal model for studies of HCMV has resulted in a number of laboratories employing experimental infection of cells in vitro to simulate both HCMV lytic and latent infection. Herein, we will focus on the techniques used in our laboratory for the isolation and use of primary cells to study aspects of HCMV latency, reactivation, and lytic infection.

Key words

  • Monocytes
  • CD34+ cells
  • Latency
  • Differentiation
  • Primary cell isolation

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  • DOI: 10.1007/978-1-62703-788-4_6
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References

  1. Smith MS, Goldman DC, Bailey AS, Pfaffle DL, Kreklywich CN, Spencer DB, Othieno FA, Streblow DN, Garcia JV, Fleming WH, Nelson JA (2010) Cell Host Microbe 8:284–291

    CAS  PubMed Central  PubMed  CrossRef  Google Scholar 

  2. Taylor-Wiedeman J, Sissons JG, Borysiewicz LK, Sinclair JH (1991) J Gen Virol 72(Pt 9):2059–2064

    CAS  PubMed  CrossRef  Google Scholar 

  3. Strobl H, Bello-Fernandez C, Riedl E, Pickl WF, Majdic O, Lyman SD, Knapp W (1997) Blood 90:1425–1434

    CAS  PubMed  Google Scholar 

  4. MacAry PA, Lindsay M, Scott MA, Craig JI, Luzio JP, Lehner PJ (2001) Proc Natl Acad Sci U S A 98:3982–3987

    CAS  PubMed Central  PubMed  CrossRef  Google Scholar 

  5. Lathey JL, Spector SA (1991) J Virol 65:6371–6375

    CAS  PubMed Central  PubMed  Google Scholar 

  6. Sallusto F, Lanzavecchia A (1994) J Exp Med 179:1109–1118

    CAS  PubMed  CrossRef  Google Scholar 

  7. Bego M, Maciejewski J, Khaiboullina S, Pari G, St Jeor S (2005) J Virol 79:11022–11034

    CAS  PubMed Central  PubMed  CrossRef  Google Scholar 

  8. Goodrum F, Reeves M, Sinclair J, High K, Shenk T (2007) Blood 110:937–945

    CAS  PubMed Central  PubMed  CrossRef  Google Scholar 

  9. Roback JD, Hillyer CD, Drew WL, Laycock ME, Luka J, Mocarski ES, Slobedman B, Smith JW, Soderberg-Naucler C, Todd DS, Woxenius S, Busch MP (2001) Transfusion 41:1249–1257

    CAS  PubMed  CrossRef  Google Scholar 

  10. Poole E, McGregor Dallas SR, Colston J, Joseph RS, Sinclair J (2011) J Gen Virol 92:1539–1549

    CAS  PubMed  CrossRef  Google Scholar 

  11. Slobedman B, Mocarski ES (1999) J Virol 73:4806–4812

    CAS  PubMed Central  PubMed  Google Scholar 

  12. Reeves MB, MacAry PA, Lehner PJ, Sissons JG, Sinclair JH (2005) Proc Natl Acad Sci U S A 102:4140–4145

    CAS  PubMed Central  PubMed  CrossRef  Google Scholar 

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Correspondence to John H. Sinclair .

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Poole, E., Reeves, M., Sinclair, J.H. (2014). The Use of Primary Human Cells (Fibroblasts, Monocytes, and Others) to Assess Human Cytomegalovirus Function. In: Yurochko, A., Miller, W. (eds) Human Cytomegaloviruses. Methods in Molecular Biology, vol 1119. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-788-4_6

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  • DOI: https://doi.org/10.1007/978-1-62703-788-4_6

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-787-7

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