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
. 2015 Jul 31;5(7):e328.
doi: 10.1038/bcj.2015.56.

IL-10 induces the development of immunosuppressive CD14(+)HLA-DR(low/-) monocytes in B-cell non-Hodgkin lymphoma

B Xiu  1 , Y Lin  2 , D M Grote  2 , S C Ziesmer  2 , M P Gustafson  3 , M L Maas  3 , Z Zhang  3 , A B Dietz  3 , L F Porrata  2 , A J Novak  2 , A-B Liang  4 , Z-Z Yang  2 , S M Ansell  2
Affiliations

IL-10 induces the development of immunosuppressive CD14(+)HLA-DR(low/-) monocytes in B-cell non-Hodgkin lymphoma

B Xiu et al. Blood Cancer J. .

Abstract

The biological role of monocytes and macrophages in B-cell non-Hodgkin lymphoma (NHL) is not fully understood. We have previously reported that monocytes from patients with B-cell NHL have an immunosuppressive CD14(+)HLA-DR(low/-) phenotype that correlates with a poor prognosis. However, the underlying mechanism by which CD14(+)HLA-DR(low/-) monocytes develop in lymphoma is unknown. In the present study, we found that interleukin (IL)-10, which is increased in the serum of patients with B-cell NHL, induced the development of the CD4(+)HLA-DR(low/-) population. Using peripheral blood samples from patients with B-cell NHL, we found that absolute numbers of CD14(+) monocytic cells with an HLA-DR(low/-) phenotype were higher than healthy controls and correlated with a higher International Prognostic Index score. IL-10 serum levels were elevated in lymphoma patients compared with controls and were associated with increased peripheral monocyte counts. Treatment of monocytes with IL-10 in vitro significantly decreased HLA-DR expression and resulted in the expansion of CD14(+)HLA-DR(low/-) population. We found that lymphoma B cells produce IL-10 and supernatants from cultured lymphoma cells increased the CD14(+)HLA-DR(low/-) population. Furthermore, we found that IL-10-induced CD14(+)HLA-DR(low/-) monocytes inhibited the activation and proliferation of T cells. Taken together, these results suggest that elevated IL-10 serum levels contribute to increased numbers of immunosuppressive CD14(+)HLA-DR(low/-) monocytes in B-cell NHL.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Absolute numbers of monocytes are increased in B-cell NHL. (a and b) A graph showing absolute monocyte counts in blood from NHL patients and healthy donors (a) or patients with different histologies (b) measured by flow cytometry. DLBCL, diffuse large B-cell lymphoma; FL, follicular lymphoma; MCL, mantle cell lymphoma; MZL, marginal zone lymphoma. One milliliter freshly drawn whole blood was stained with a panel of Abs and analyzed by flow cytometry. Absolute monocyte counts were calculated as the numbers of CD14+ cells per microliter of blood (NHL: n=22, Ctrl: n=26). (c) Representative plots showing coexpression of CD14 and CD16 in blood from a healthy donor. Classical monocytes: CD14++CD16−; intermediate monocytes: CD14++CD16+; non-classical monocytes: CD14+CD16++. (d) Graphs showing the absolute counts (upper panel) or percentages (lower panel) of classical, intermediate and non-classical monocytes (Mo) in blood from NHL patients and healthy donors measured by flow cytometry.
Figure 2
Figure 2
Monocytes from B-cell NHL exhibit CD14+HLA-DRlow/− phenotype. (a) Representative plots showing coexpression of CD14 and HLA-DR in blood from healthy donors (Ctrl) and lymphoma patients (NHL). CD14+HLA-DRlow/− cells were defined based on isotype control gate. (b) Graphs showing absolute counts (left) or the percentage (right) of CD14+HLA-DRlow/− cells in blood from NHL patients and healthy donors measured by flow cytometry. (c) Graphs showing expression level of HLA-DR on classical, intermediate and non-classical monocytes in blood from NHL patients and healthy donors. (d) A graph showing the percentage of CD14+HLA-DRlow/− monocytes in blood from NHL patients with different histologies. (e) Graphs showing the percentage of CD14+HLA-DRlow/− monocytes in blood from NHL patients with different International Prognostic Index (IPI) scores.
Figure 3
Figure 3
Phenotype of CD14+HLA-DRlow/− or HLA-DR+ monocytes in B-cell NHL. A 10-color flow cytometry was employed to determine the expression of surface markers included CD86, CD64, CD32, CD80, CD142, TNFR2, CD40, B7-H1, PD-1, CD206, CD163 and CD169 on CD14+HLA-DRlow/− or CD14+HLA-DR+ monocytes from both healthy donors (NC) or lymphoma patients (NHL).
Figure 4
Figure 4
IL-10 induces the development of CD14+HLA-DRlow/− monocytes. (a) Representative plots showing the expression of HLA-DR on CD14+ monocytes treated with or without IL-10 or IFN-γ in the presence or absence of M-CSF for 24 h. (b) Summarization of HLA-DR expression level in CD14+ monocytes treated with or without IL-10, IFN-γ, IL-4, M-CSF or GM-CSF. (c) Graph showing the percentage of HLA-DRlow/− cells of CD14+ monocytes treated with IL-10 at escalating doses. (d) Graph showing serum IL-10 level in healthy donors or patients with B-cell NHL. IL-10 concentration was measured by multiplex ELISA (Luminex). (e) Graph showing absolute monocyte counts (AMC)/μl in lymphoma patients with undetectable (<20 pg/ml, n=118) or detectable (>20 pg/ml, n=101) IL-10 serum levels. (f) Representative histograms showing the expression of IL-10Rα or IL-10Rβ on CD14+ monocytes in patients with B-cell NHL.
Figure 5
Figure 5
Lymphoma cells produce IL-10 and induce the development of CD14+HLA-DRlow/− monocytes. (a and b) Graphs showing IL-10 concentration in culture supernatant of MNCs from lymphoma tissues (a) or lymphoma cell lines (b). IL-10 concentration was measured by ELISA. (c and d) Representative plots showing HLA-DR expression on CD14+ cells cultured with or without supernatant of lymphoma cells (c) or SuDHL-2 cells (d). (e) Representative plots showing HLA-DR expression on CD14+ cells treated with or without IL-10 in the presence of αIL-10R Ab or isotype control. The percentage of CD14+HLA-DRlow/− cells from above experiment setting was summarized in the graph below (n=3). (f) Graph showing the percentage of HLA-DRlow/− cells in CD14+ monocytes cultured with or without supernatant from SuDHL-2 cells in the presence of αIL-10R Ab or isotype control (n=3).
Figure 6
Figure 6
IL-10-treated monocytes inhibit T-cell activation and proliferation. (a) Representative plots showing the expression of CD69 and CD25 on activated CD3+ T cells (Ta) cocultured with or without untreated or IL-10-treated monocytes (Mo) for 3 days. Expression of CD69 and CD25 on resting T cells (Tr) was measured and used as a control. (b) Summarization of CD69 or CD25 induction in Tr or Ta cocultured with or without untreated or IL-10-treated Mo for 3 days, n=5. (c and d) Representative histograms showing proliferation measured by CFSE staining of Tr or Ta cocultured with or without untreated or IL-10-treated Mo for 3 or 6 days. Proliferative capacity was expressed by calculating the number of CFSEdim cells. The results from multiple experiments were summarized in panel (d). The percentage change of CFSEdim cells in each group was expressed as fold change when compared with the group Ta alone, n=5. (e) Graph showing the percentage change of CFSEdim T cells treated with or without untreated or IL-10-treated Mo in the presence of αIL-10R Ab or isotype control for 3 or 6 days, n=2.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Rosenquist R, Davi F, Ghia P. The microenvironment in lymphomas—dissecting the complex crosstalk between tumor cells and 'by-stander' cells. Semin Cancer Biol. 2014;24:1–2. - PubMed
    1. Yang ZZ, Ansell SM. The tumor microenvironment in follicular lymphoma. Clin Adv Hematol Oncol. 2012;10:810–818. - PubMed
    1. Juszczynski P, Nowak J, Warzocha K. Host immune response in B-cell lymphomas: friend or foe. Arch Immunol Ther Exp (Warsz) 2008;56:245–255. - PubMed
    1. Scott DW, Gascoyne RD. The tumour microenvironment in B cell lymphomas. Nat Rev Cancer. 2014;14:517–534. - PubMed
    1. Wahlin BE, Sander B, Christensson B, Kimby E. CD8+ T-cell content in diagnostic lymph nodes measured by flow cytometry is a predictor of survival in follicular lymphoma. Clin Cancer Res. 2007;13 (2 Pt 1:388–397. - PubMed

Publication types