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
. 2018 Jan 29;13(1):e0191482.
doi: 10.1371/journal.pone.0191482. eCollection 2018.

Cytomegalovirus induces HLA-class-II-restricted alloreactivity in an acute myeloid leukemia cell line

Michael Koldehoff  1 , Monika Lindemann  2 , Stefan R Ross  3 , Ahmet H Elmaagacli  1   4
Affiliations

Cytomegalovirus induces HLA-class-II-restricted alloreactivity in an acute myeloid leukemia cell line

Michael Koldehoff et al. PLoS One. .

Abstract

Cytomegalovirus (HCMV) reactivation is found frequently after allogeneic hematopoietic stem cell transplantation (alloSCT) and is associated with an increased treatment-related mortality. Recent reports suggest a link between HCMV and a reduced risk of cancer progression in patients with acute leukemia or lymphoma after alloSCT. Here we show that HCMV can inhibit the proliferation of the acute myeloid leukemia cell line Kasumi-1 and the promyeloid leukemia cell line NB4. HCMV induced a significant up-regulation of HLA-class-II-molecules, especially HLA-DR expression and an increase of apoptosis, granzyme B, perforin and IFN-γ secretion in Kasumi-1 cells cocultured with peripheral blood mononuclear cells (PBMCs). Indolamin-2,3-dioxygenase on the other hand led only to a significant dose-dependent effect on IFN-γ secretion without effects on proliferation. The addition of CpG-rich oligonucleotides and ganciclovir reversed those antiproliferative effects. We conclude that HCMV can enhance alloreactivity of PBMCs against Kasumi-1 and NB4 cells in vitro. To determine if this phenomenon may be clinically relevant further investigations will be required.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1
A: HCMV infection inhibits proliferation of AML cells. 12,500–400,000 AML cells were cultured for six days in the absence or presence of HCMV infection. Proliferation was measured by H3 thymidine incorporation. Mean and SD of counts per minute (CPM) are presented. B: Apoptosis of HCMV-infected AML cells. The rate of apoptosis in AML cells 48 h without vs. with HCMV-infection and apoptosis in AML cells co-cultured with PBMC measured by annexin detection (%). C: Proliferation of HCMV-infected Kasumi-1 cells and co-culture experiments of Kasumi-1 and PBMC. Proliferation of Kasumi-1 cells without vs. with HCMV infection and co-cultured with PBMC was analyzed by H3 thymidine uptake and results were given as counts per minute (CPM).
Fig 2
Fig 2
A: Flow cytometry analysis of Kasumi-1 cells and PBMC. Expression of HLA class I and class II on Kasumi-1 cells without vs. with HCMV infection, both co-cultured with PBMC, was analyzed by flow cytometry. MFI, median fluorescence intensity, (w/o), without. B: HLA class II analysis of Kasumi-1 cells and PBMC. Expression of HLA-DP, HLA-DQ and HLA-DR on Kasumi-1 cells without vs. with HCMV infection, both co-cultured with PBMC, was analyzed by flow cytometry.
Fig 3
Fig 3
A: Secretion of IFN-α in co-culture experiments of Kasumi-1 cells. Panel (A) to (D) display co-culture experiments of Kasumi-1 cells without vs. with HCMV infection and PBMC of healthy controls. IFN-α (panel A) was determined by ELISpot assay. ELISpot results are given as spots per well. Kasumi-1 cells without and with HCMV infection were compared by Wilcoxon matched pairs test, Kasumi-1 cells and Kasumi-1 cells plus PBMC were compared by Mann-Whitney test. B: Secretion of IFN-γ in co-culture experiments of Kasumi-1 cells. IFN-γ (panel B) was determined by ELISpot assay. C: Secretion of granzyme B in co-culture experiments of Kasumi-1 cells. Granzyme B (panel C) was determined by ELISpot assay. D: Secretion of perforin in co-culture experiments of Kasumi-1 cells. Perforin (panel D) was determined by ELISpot assay, (w/o), without.
Fig 4
Fig 4
A: Alloreactivity-mediated proliferation of indolamin-2,3-dioxygenase-treated HCMV-infected AML cells co-cultured with mixed lymphocyte cultures. HCMV-infected AML cells co-cultured with mixed lymphocyte cultures from healthy controls were performed without and with 1 μM and 10 μM indolamin-2,3-dioxygenase (IDO). Panel (A) shows proliferative responses determined by H3 thymidine uptake (n = 3 experiments) and proliferative responses are presented as counts per minute (CPM). B: IFN-γ secretion by indolamin-2,3-dioxygenase-treated HCMV-infected AML cells co-cultured with mixed lymphocyte cultures. Panel (B) displays IFN-γ secretion determined by ELISpot, ELISpot results are given as spots per well. One-way-ANOVA (Friedman test) indicates that the groups were significantly different (p = 0.008) in terms of IFN-γ secretion.
Fig 5
Fig 5. Effect of CpG and ganciclovir on proliferation of HCMV-infected leukemia cells.
Kasumi-1 or NB4 cells without (w/o) and with HCMV infection were co-cultured with PBMC without and with CpG-rich ODNs and/or ganciclovir and cell proliferation was measured by H3 thymidine uptake. The addition of CpG-rich ODNs to the co-cultures resulted in significantly higher proliferation of Kasumi-1 cells with vs. without CMV infection (panel A). Ganciclovir and the combination of CpG-rich ODNs and ganciclovir led to significantly increased proliferation in HCMV infected NB4 cells (panel B). Proliferative responses are given as counts per minute (CPM). Kasumi-1 or NB4 cells without and with HCMV infection were compared by Wilcoxon matched pairs test.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Boeckh M, Geballe AP. Cytomegalovirus: pathogen, paradigm, and puzzle. J Clin Invest. 2011; 121: 1673–1680. doi: 10.1172/JCI45449 - DOI - PMC - PubMed
    1. Drylewicz J, Schellens IM, Gaiser R, Nanlohy NM, Quakkelaar ED, Otten H, et al. Rapid reconstitution of CD4 T cells and NK cells protects against CMV-reactivation after allogeneic stem cell transplantation. J Transl Med. 2016; 14: 230–239. doi: 10.1186/s12967-016-0988-4 - DOI - PMC - PubMed
    1. Lisboa LF, Tong Y, Kumar D, Pang XL, Asberg A, Hartmann A, et al. Analysis and clinical correlation of genetic variation in cytomegalovirus. Transpl Infect Dis. 2012; 14:132–140. doi: 10.1111/j.1399-3062.2011.00685.x - DOI - PubMed
    1. Schmidt-Hieber M, Labopin M, Beelen D, Volin L, Ehninger G, Finke J, et al. CMV serostatus still has an important prognostic impact in de novo acute leukemia patients after allogeneic stem cell transplantation: a report from the Acute Leukemia Working Party of EBMT. Blood. 2013; 122: 3359–3364. doi: 10.1182/blood-2013-05-499830 - DOI - PubMed
    1. Einsele H, Ehninger G, Steidle M, Fischer I, Bihler S, Gerneth F, et al. Lymphocytopenia as an unfavorable prognostic factor in patients with cytomegalovirus infection after bone marrow transplantation. Blood. 1993; 82: 1672–1678. - PubMed

Publication types

MeSH terms

Substances

Grants and funding

All materials, experiments, financial and administrative support for this study were funded by the academic budget of Michael Koldehoff and Ahmet H. Elmaagacli.