"VSports在线直播" 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 VSports app下载. 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. V体育官网.

. 2010 Nov;40(11):3097-106.
doi: 10.1002/eji.201040659. Epub 2010 Oct 19.

VSports注册入口 - Granulocyte-colony stimulating factor drives the in vitro differentiation of human dendritic cells that induce anergy in naïve T cells

Maura Rossetti  1 , Silvia Gregori, Maria Grazia Roncarolo
Affiliations
Free PMC article

Granulocyte-colony stimulating factor drives the in vitro differentiation of human dendritic cells that induce anergy in naïve T cells

"VSports在线直播" Maura Rossetti et al. Eur J Immunol. 2010 Nov.
Free PMC article

Abstract (VSports)

G-CSF is a modulator of T-cell and DC functions. Previous reports show that monocytes from G-CSF-treated (post-G) healthy donors differentiate into tolerogenic DC in vitro in the presence of autologous serum, containing high levels of IL-10 and IFN-α, and in turn induce type 1 Treg (Tr1) cells. However, the direct effect of G-CSF on DC differentiation was not investigated. Here, we show that monocytes differentiated in the presence of exogenous G-CSF (G-DC) remain CD14(+) CD1a(-) , but acquire a DC-like morphology, express CD83 and CD86 and low levels of the tolerogenic markers Ig-like transcript (ILT)4 and HLA-G. G-DC spontaneously produce IL-10 and, upon stimulation, low levels of IL-12. G-DC display low stimulatory capacity and induce anergy in naïve T cells, but do not confer suppressive function. Therefore, in vitro differentiation of monocyte-derived DC in the presence of G-CSF can replicate some but not all features of post-G DC VSports手机版. These findings indicate that the tolerogenic properties of G-CSF do not exclusively reside in its direct effect on DC, which in turn induce T-cell anergy, but also in its ability to generate a tolerogenic milieu in vivo, which is necessary for Tr1 cell induction and cannot be replicated in vitro. .

PubMed Disclaimer

Figures

Figure 1
Figure 1
G-CSF drives the differentiation of DC-like cells similar to post-G DC. MoDC were differentiated by 7-day culture in the presence of G-CSF and IL-4 (G-DC), or GM-CSF and IL-4 (iDC), in some cases with the addition of LPS during the last 2 days of culture (mDC). (A) Cytospins were performed by centrifugation of 105 cells onto slides, followed by staining with May Grünwald-Giemsa (magnification 60×). One representative donor out of four tested in two independent experiments is shown. (B–D) Expression of CD14, CD1a, CD16 (B), HLA-DR, CD80, CD83, CD86 (B and C), ILT2, ILT3, ILT4 and HLA-G (D) was evaluated by FACS on day 7. A representative donor and the mean+SEM of at least ten donors tested in five independent experiments are presented. *p<0.05, **p<0.01, ***p<0.001; Mann–Whitney test.
Figure 2
Figure 2
G-DC spontaneously produce IL-10 and upon activation secrete low levels of IL-12. MoDC were differentiated by 7-day culture in the presence of G-CSF and IL-4 (G-DC), or GM-CSF and IL-4 (iDC), in some cases with the addition of LPS during the last 2 days of culture (mDC). (A and B) DC were washed and seeded in the absence (A) or presence (B) of LPS and IFN-γ; supernatants were collected 48 h later to measure cytokine release. Data show mean+SEM of at least 12 donors tested in six independent experiments. *p<0.05, **p<0.01; Mann–Whitney test.
Figure 3
Figure 3
G-DC display hypo-stimulatory capacity and promote T-cell anergy. Naïve CD4+ T cells were primed with irradiated (6000 rad) allogeneic G-DC, iDC, or mDC at 10:1 ratio. (A) After 4 days, 50 μL of supernatant were taken to test IFN-γ release (right) and cells were pulsed for 16 h with 1 μCi/well 3H-thymidine (left). Data show mean+SEM of 11 donors for proliferation and 13 donors for IFN-γ production, tested in seven independent experiments. (B) Naïve CD4+ T cells were cultured with irradiated allogeneic DC or CD14+ monocytes at 1:1 ratio. After 4 days, cells were pulsed for 16 h with 1 μCi/well 3H-thymidine. Data show mean+SEM of three donors. (C) After 14 days of culture, T cells primed with G-DC [T(G-DC)], iDC [T(iDC)], or mDC [T(mDC)] were washed and evaluated for their proliferative response to mDC from the same allogeneic donor. After 2 days, 50 μL of supernatant were taken to test IFN-γ release (right) and cells were pulsed for 16 h with 1 μCi/well 3H-thymidine (left). Data show mean+SEM of 14 donors tested in eight independent experiments. *p<0.05, **p<0.01, ***p<0.001; Mann–Whitney test.
Figure 4
Figure 4
T cells primed with G-DC acquire a Tr1-like cytokine profile but not suppressive capacity. Naïve CD4+ T cells were cultured with irradiated (6000 rad) allogeneic G-DC [T(G-DC)] or mDC [T(mDC)] at 10:1 ratio. (A) After 14 days, T-cell lines were washed and re-stimulated with mDC from the same allogeneic donor. Culture supernatants were collected at 24 h (IL-2) and 48 h (IL-4, IL-5, IL-10, IL-17, IFN-γ and TNF-α), and cytokine levels were evaluated by Bioplex. Data show mean+SEM of eight donors tested in four independent experiments. (B) After 14 days of culture, T-cell lines were washed and evaluated for their ability to suppress the proliferation of autologous CD4+ T cells activated by mDC from the same allogeneic donor (MLC). After 3 days, 50 μL of supernatant were taken to test IFN-γ release (middle and right) and cells were pulsed for 16 h with 1 μCi/well 3H-thymidine (left). Data show mean+SEM of five donors for proliferation and eight donors for IFN-γ production, tested in three to six independent experiments (left and middle), and the production of IFN-γ by the four suppressive donors (right). *p<0.05, **p<0.01, *** p<0.001; Mann–Whitney test. ND: normal donor.
Figure 5
Figure 5
Addition of IL-10, anti-IL-12 and anti-TNF-α antibodies does not rescue the suppressive ability of T(G-DC) cells. Naïve CD4+ T cells were cultured with allogeneic G-DC at 10:1 ratio, in the absence [T(G-DC)] or presence [T(G-DC)+IL-10] of exogenous IL-10 and blocking antibodies against IL-12 and TNF-α. (A) After 14 days of culture, T-cell lines were washed and evaluated for their proliferative response to mDC from the same allogeneic donor. After 2 days, 50 μL of supernatant were taken to test IFN-γ release (right) and cells were pulsed for 16 h with 1 μCi/well 3H-thymidine (left). Data show mean+SEM of five donors for proliferation and seven donors for IFN-γ production, tested in three to four independent experiments. (B) After 14 days of culture, T-cell lines were washed and evaluated for their ability to suppress the proliferation of autologous CD4+ T cells activated by mDC from the same allogeneic donor (MLC). After 3 days, 50 μL of supernatant were taken to test IFN-γ release (right) and cells were pulsed for 16 h with 1 μCi/well 3H-thymidine (left). Data show mean+SEM of three donors tested in two independent experiments.
See this image and copyright information in PMC

References

    1. Demetri GD, Griffin JD. Granulocyte colony-stimulating factor and its receptor. Blood. 1991;78:2791–2808. - PubMed
    1. Liongue C, Wright C, Russell AP, Ward AC. Granulocyte colony-stimulating factor receptor: stimulating granulopoiesis and much more. Int. J. Biochem. Cell Biol. 2009;41:2372–2375. - PubMed
    1. Sloand EM, Kim S, Maciejewski JP, Van Rhee F, Chaudhuri A, Barrett J, Young NS. Pharmacologic doses of granulocyte colony-stimulating factor affect cytokine production by lymphocytes in vitro and in vivo. Blood. 2000;95:2269–2274. - PubMed
    1. Franzke A, Piao W, Lauber J, Gatzlaff P, Konecke C, Hansen W, Schmitt-Thomsen A, et al. G-CSF as immune regulator in T cells expressing the G-CSF receptor: implications for transplantation and autoimmune diseases. Blood. 2003;102:734–739. - PubMed
    1. Morikawa K, Morikawa S, Nakamura M, Miyawaki T. Characterization of granulocyte colony-stimulating factor receptor expressed on human lymphocytes. Br. J. Haematol. 2002;118:296–304. - PubMed (V体育官网入口)

Publication types

MeSH terms