Growth of human cytomegalovirus in primary macrophages

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

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

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
Volume16
Issue number1
DOIs
StatePublished - Sep 1998

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Macrophages
Cytomegalovirus
Growth
Viruses
Dendritic Cells
Granulocyte-Macrophage Colony-Stimulating Factor
Cytokines
Transplants
T-cells
Cytomegalovirus Infections
Pathogens
Virus Replication
Infection
Interleukin-1
Cell culture
Interleukin-4
Interferons
Interleukin-2
Monocytes
Bone

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Growth of human cytomegalovirus in primary macrophages. / Söderberg-Nauclér, Cecilia; Fish, Kenneth N.; Nelson, Jay.

In: Methods: A Companion to Methods in Enzymology, Vol. 16, No. 1, 09.1998, p. 126-138.

Research output: Contribution to journalArticle

Söderberg-Nauclér, Cecilia ; Fish, Kenneth N. ; Nelson, Jay. / Growth of human cytomegalovirus in primary macrophages. In: Methods: A Companion to Methods in Enzymology. 1998 ; Vol. 16, No. 1. pp. 126-138.
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