Growth of human cytomegalovirus in primary macrophages

Cecilia Söderberg-Nauclér, Kenneth N. Fish, Jay A. Nelson

Research output: Contribution to journalArticlepeer-review

38 Scopus citations


Human cytomegalovirus (HCMV) is a major human pathogen that causes considerable disease among immunocompromised individuals. A primary infection results in life-long persistence of the virus in a latent form. HCMV is known to be transferred by blood products, bone marrow, and solid organs, but the cell type that carries the latent infection has been difficult to identify. We have recently demonstrated reactivation of latent HCMV in allogeneically stimulated monocyte-derived macrophages (Allo-MDM). Reactivation occurred only in macrophages produced by allogeneic but not mitogenic stimulation. The presence of dendritic cell markers on some Allo-MDM cells suggested that these macrophages were related to dendritic cells. However, dendritic cells obtained by stimulation of monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony stimulating factor (GM-CSF) were not permissive for HCMV infection. The cellular and cytokine components which are essential for HCMV replication and reactivation of virus were also examined in Allo-MDM. The importance of both CD4- or CD8-positive T cells in the generation of HCMV permissive Allo-MDM was demonstrated by negative selection or blocking experiments using antibodies directed against both HLA class I and HLA class II molecules. Examination of the cytokines essential for the generation of HCMV permissive Allo-MDM identified γ-interferon (IFN-γ, but not IL-1, IL-2, tumor necrosis factor α, or GM-CSF as critical components in the generation of these macrophages. However, addition of IFN-γ to unstimulated macrophage cultures was insufficient to reactivate virus. These results indicate the importance of a specific moncyte stimulus in the generation of a unique HCMV permissive macrophage phenotype as well as why virus is commonly reactivated in transplant patients.

Original languageEnglish (US)
Pages (from-to)126-138
Number of pages13
JournalMethods: A Companion to Methods in Enzymology
Issue number1
StatePublished - Sep 1998

ASJC Scopus subject areas

  • Molecular Biology
  • General Biochemistry, Genetics and Molecular Biology


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