Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Sep 16;8(3):284-91.
doi: 10.1016/j.chom.2010.08.001.

Granulocyte-colony stimulating factor reactivates human cytomegalovirus in a latently infected humanized mouse model

M Shane Smith  1 , Devorah C Goldman, Alexis S Bailey, Dana L Pfaffle, Craig N Kreklywich, Doran B Spencer, Florence A Othieno, Daniel N Streblow, J Victor Garcia, William H Fleming, Jay A Nelson
Affiliations

Granulocyte-colony stimulating factor reactivates human cytomegalovirus in a latently infected humanized mouse model

M Shane Smith et al. Cell Host Microbe. .

Abstract

Human cytomegalovirus (HCMV) is a significant cause of morbidity and mortality in organ transplant recipients. The use of granulocyte-colony stimulating factor (G-CSF)-mobilized stem cells from HCMV seropositive donors is suggested to double the risk of late-onset HCMV disease and chronic graft-versus-host disease in recipients when compared to conventional bone marrow transplantation with HCMV seropositive donors, although the etiology of the increased risk is unknown. To understand mechanisms of HCMV transmission in patients receiving G-CSF-mobilized blood products, we generated a NOD-scid IL2Rγ(c)(null)-humanized mouse model in which HCMV establishes latent infection in human hematopoietic cells. In this model, G-CSF induces the reactivation of latent HCMV in monocytes/macrophages that have migrated into organ tissues. In addition to establishing a humanized mouse model for systemic and latent HCMV infection, these results suggest that the use of G-CSF mobilized blood products from seropositive donors pose an elevated risk for HCMV transmission to recipients.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Mobilization of monocytes to the peripheral blood in HCMV-infected, huCD34+ engrafted mice with G-CSF and AMD3100. a, Peripheral blood mononuclear cells from HCMV-infected, engrafted mice were analyzed for murine CD45 (Ly5.2), huCD45, huCD33, and huCD14 expression by flow cytometry before and after 7 days of mobilization with either G-CSF or G-CSF + AMD3100. b,c, AMD3100 + G-CSF treatment resulted in an earlier and more robust mobilization of total huCD45+ PBMCs and huCD33+/huCD14+ monocytes in comparison to G-CSF treatment alone. Total huCD45+ cells/µl (b) or huCD33+/huCD14+ cells/µl (c) of peripheral blood in HCMV-infected engrafted mice were determined pre-mobilization and at days 3 and 7 of mobilization with either G-CSF or G-CSF + AMD3100 by normalizing the percent of cells positive for huCD45 expression by flow cytometry to total white blood cell counts. formula image, pre-mobilized; formula image, mobilized. Data represent mean ± s.e.m. (n=4). *, P < 0.05.
Fig. 2
Fig. 2
HSC mobilization increases viral genomic DNA in organ tissues of huCD34+ engrafted mice infected with HCMV. a–c, G-CSF treatment of HCMV-infected engrafted mice for 7 days results in increased HCMV genomic DNA in spleen (a), bone marrow mononuclear cells (b), and liver (c). Engrafted mice were injected IP with mock-infected fibroblasts (engrafted mock), UV-inactivated HCMV-treated fibroblasts (engrafted UV-HCMV), or HCMV-infected fibroblasts (engrafted HCMV). At 4 weeks post-infection, engrafted mice were treated with G-CSF (+ G-CSF) for 7 days or left untreated (− G-CSF) and sacrificed. As an additional negative control, non-engrafted mice were injected IP with HCMV-infected fibroblasts, treated with G-CSF for 7 days at 4 weeks post-infection, and sacrificed (Non-engrafted HCMV + G-CSF). Total DNA was harvested from organ tissue and analyzed for HCMV genomic DNA by quantitative real time PCR. Data represent mean copies/µg ± s.d. (n=5). *, P < 0.05. d, Enhanced HSC mobilization with AMD3100 + G-CSF correlates with a further increase in HCMV viral load in liver. HCMV-infected engrafted mice were administered PBS, G-CSF, or AMD3100 + G-CSF for 7 days and sacrificed. Total DNA was isolated from liver and analyzed by quantitative real time PCR for HCMV genomic DNA. Data represent mean copies/µg ± s.d. (n=3, HCMV + PBS; n=4, HCMV + G-CSF, HCMV + G-CSF + AMD3100). *, P < 0.05.
Fig. 3
Fig. 3
HCMV permissively infects monocytes/macrophages in huCD34+ engrafted mice. a, HCMV mRNAs are expressed in liver following HSC mobilization. Engrafted mice were infected with HCMV, treated with G-CSF (n=5) or PBS (n=3) for 7 days at 4 weeks post-infection, and sacrificed at 6 weeks post-infection. Levels of UL55 (gB, formula image), UL83 (pp65, formula image), and UL146 (formula image) mRNAs were quantified by real-time PCR of total liver and bone marrow mononuclear cell mRNA. PBS AVG Liver, PBS AVG BM, and G-CSF AVG BM represent the mean values of transcripts for PBS-treated mouse liver tissue, PBS-treated mouse bone marrow mononuclear cells, and G-CSF-treated mouse bone marrow mononuclear cells respectively. b,c, HCMV genomic DNA is present in the bone marrow of G-CSF-mobilized and PBS-treated mice (b) and increases in the liver of HCMV-infected engrafted mice following G-CSF treatment (c). Engrafted mice were infected with HCMV, treated with G-CSF (n=5) or PBS (n=3) for 7 days at 4 weeks post-infection, and sacrificed at 6 weeks post-infection. Total DNA was harvested from organ tissue and analyzed for HCMV genomic DNA by quantitative real time PCR. Data represent mean copies/µg ± s.d. (n=5). *, P < 0.01. d, HCMV gB/gH expression is limited to CD14+ HLA-DR+ monocytes. Engrafted mice were injected IP with HCMV-infected fibroblasts, UV-inactivated HCMV-treated fibroblasts, or mock-infected fibroblasts. At 4 weeks post-infection, mice were administered G-CSF for 7 days and sacrificed. Liver tissue sections were stained with antibodies against huCD14, huHLA-DR, and HCMV gB/gH. 4-color immunofluorescence images of 8 µm liver cryosections were obtained for Hoechst-stained nuclei (blue; all panels), huCD14 (green; left), huHLA-DR (red; middle), and HCMV gB/gH (purple; right). HLA-DR+CD14+ monocytes co-expressing HCMV gB and gH were seen only in HCMV-infected mice treated with G-CSF. Scale bars = 25µm.
Fig. 3
Fig. 3
HCMV permissively infects monocytes/macrophages in huCD34+ engrafted mice. a, HCMV mRNAs are expressed in liver following HSC mobilization. Engrafted mice were infected with HCMV, treated with G-CSF (n=5) or PBS (n=3) for 7 days at 4 weeks post-infection, and sacrificed at 6 weeks post-infection. Levels of UL55 (gB, formula image), UL83 (pp65, formula image), and UL146 (formula image) mRNAs were quantified by real-time PCR of total liver and bone marrow mononuclear cell mRNA. PBS AVG Liver, PBS AVG BM, and G-CSF AVG BM represent the mean values of transcripts for PBS-treated mouse liver tissue, PBS-treated mouse bone marrow mononuclear cells, and G-CSF-treated mouse bone marrow mononuclear cells respectively. b,c, HCMV genomic DNA is present in the bone marrow of G-CSF-mobilized and PBS-treated mice (b) and increases in the liver of HCMV-infected engrafted mice following G-CSF treatment (c). Engrafted mice were infected with HCMV, treated with G-CSF (n=5) or PBS (n=3) for 7 days at 4 weeks post-infection, and sacrificed at 6 weeks post-infection. Total DNA was harvested from organ tissue and analyzed for HCMV genomic DNA by quantitative real time PCR. Data represent mean copies/µg ± s.d. (n=5). *, P < 0.01. d, HCMV gB/gH expression is limited to CD14+ HLA-DR+ monocytes. Engrafted mice were injected IP with HCMV-infected fibroblasts, UV-inactivated HCMV-treated fibroblasts, or mock-infected fibroblasts. At 4 weeks post-infection, mice were administered G-CSF for 7 days and sacrificed. Liver tissue sections were stained with antibodies against huCD14, huHLA-DR, and HCMV gB/gH. 4-color immunofluorescence images of 8 µm liver cryosections were obtained for Hoechst-stained nuclei (blue; all panels), huCD14 (green; left), huHLA-DR (red; middle), and HCMV gB/gH (purple; right). HLA-DR+CD14+ monocytes co-expressing HCMV gB and gH were seen only in HCMV-infected mice treated with G-CSF. Scale bars = 25µm.
See this image and copyright information in PMC

Comment in

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Aiuti A, Webb IJ, Bleul C, Springer T, Gutierrez-Ramos JC. The chemokine SDF-1 is a chemoattractant for human CD34+ hematopoietic progenitor cells and provides a new mechanism to explain the mobilization of CD34+ progenitors to peripheral blood. J Exp Med. 1997;185:111–120. - PMC - PubMed
    1. Anderson D, DeFor T, Burns L, McGlave P, Miller J, Wagner J, Weisdorf D. A comparison of related donor peripheral blood and bone marrow transplants: importance of late-onset chronic graft-versus-host disease and infections. Biol Blood Marrow Transplant. 2003;9:52–59. - PubMed
    1. Asano-Mori Y, Kanda Y, Oshima K, Kako S, Shinohara A, Nakasone H, Sato H, Watanabe T, Hosoya N, Izutsu K, et al. Clinical features of late cytomegalovirus infection after hematopoietic stem cell transplantation. Int J Hematol. 2008;87:310–318. - PubMed
    1. Bentz GL, Jarquin-Pardo M, Chan G, Smith MS, Sinzger C, Yurochko AD. Human cytomegalovirus (HCMV) infection of endothelial cells promotes naive monocyte extravasation and transfer of productive virus to enhance hematogenous dissemination of HCMV. J Virol. 2006;80:11539–11555. - PMC - PubMed
    1. Bidanset DJ, Rybak RJ, Hartline CB, Kern ER. Replication of human cytomegalovirus in severe combined immunodeficient mice implanted with human retinal tissue. J Infect Dis. 2001;184:192–195. - PubMed

Publication types

MeSH terms