J Bacteriol Virol.  2008 Mar;38(1):39-46. 10.4167/jbv.2008.38.1.39.

Human Cytomegalovirus Induces Intercellular Adhesion Molecule-1 Expression in a Monocytic Cell Line, THP-1

• Kim MS
• Yi HA
• Lee CH

• KMID: 2168517
• DOI: http://doi.org/10.4167/jbv.2008.38.1.39

Abstract

It has been reported that inflammatory diseases such as pneumonitis, retinitis, and hepatitis are associated with human cytomegalovirus (HCMV). Intercellular adhesion molecule (ICAM)-1 is an important inflammatory mediator, helping monocytes adhere to endothelial cells when tissues are infected by pathogen including the HCMV. However, little is known about the mechanism of ICAM-1 stimulation by the HCMV infection in monocytes. In this study, a monocytic cell line THP-1 was used to understand ICAM-1 expression by the HCMV infection. Flow cytometric analyses demonstrated that ICAM-1 was stimulated by the HCMV in THP-1 cells with maximum at 24 hours post infection. The stimulated ICAM-1 expression was dependent on the amount of input virus. In order to understand the mechanism of ICAM-1 stimulation during the HCMV infection, cells were treated with specific inhibitors of key elements in inflammation: NF-kappaB inhibitor PDTC, cyclooxygenase 2 inhibitor NS398, and MEK inhibitor PD98059. Flow cytometric analyses revealed that ICAM-1 expression was decreased when treated with PDTC, but not with NS398 or PD98059. Thus, it is suggested that HCMV-induced ICAM-1 expression in THP-1 cells is associates with NF-kappaB.

Keyword

HCMV; Inflammation; ICAM-1; NF-kappaB

MeSH Terms

Cell Line
Cyclooxygenase 2
Cytomegalovirus
Endothelial Cells
Flavonoids
Hepatitis
Humans
Inflammation
Intercellular Adhesion Molecule-1
Monocytes
NF-kappa B
Nitrobenzenes
Pneumonia
Proline
Retinitis
Sulfonamides
Thiocarbamates
Viruses
Cyclooxygenase 2
Flavonoids
Intercellular Adhesion Molecule-1
NF-kappa B
Nitrobenzenes
Proline
Sulfonamides
Thiocarbamates

Figure

• 

  Figure 1. Stimulation of ICAM-1 expression by the HCMV infection on the surface of THP-1 cells. THP-1 cells were infected by HCMV TB40/E at MOI of 1 pfu/cell. Cells were harvested at indicated times and stained with PE-conjugated anti-ICAM-1 antibody. PE fluorescence was determined by a flow cytometry. (A) Flow cytometrogram of a representative experiment. C, background fluorescence; M, mock-infected cells; V, virus-infected cells. (B) Time course of ICAM-1 expression. Data are averages of three or more experiments with error bars. Open circles, mock-infected cells; closed circles, HCMV-infected cells.

• 

  Figure 2. HCMV MOI-dependent stimulation of ICAM-1 expression. THP-1 cells were infected with HCMV TB40/E at different MOIs. Cells were harvested at 24 hours post infection. Cells were stained with PE-conjugated anti-ICAM-1 antibody and analyzed by a flow cytometry. (A) Flow cytometrogram. C, background fluorescence; V, virus-infected cells. (B) Relative ICAM-1 expression.

• 

  Figure 3. Effect of specific inhibitors on the HCMV-induced ICAM-1 expression. Cells were pre-treated with inhibitors for 30 minutes and infected with HCMV TB40/E at 1 MOI. Twenty four hours after infection, cells were harvested, fixed and stained with PE-conjugated anti-ICAM-1 antibody and fluorescence was determined by a flow cytometry. (A) treated with NF-κB inhibtor PDTC (1 μM); (B) treated with COX-2 inhibitor NS398 (50 μM); (C) treated with MEK inhibtor PD98059 (50 μM).

• 

  Figure 4. HCMV does not stimulate COX-2 expression or ERK1/2 phosphorylation in THP-1 cells. THP-1 or HFF cells were infected with HCMV (strain TB40/E, MOI = 1). At indicated times after virus infection, cells were harvested and subjected to western blot analysis. (A) COX-2; (B) ERK1/2.

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