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Production of TGF-beta1 as a Mechanism for Defective Antigen-presenting Cell Function of Macrophages Generated in vitro with M-CSF.

Lee JK, Lee YR, Lee YH, Kim K, Lee CK - Immune Netw (2009)

Bottom Line: Production of TGF-beta1 by BM-Mp was confirmed by neutralization experiments of TGF-beta1 as well as by real time-polymerase chain reaction (PCR).Quantitative real time-PCR analysis also confirmed the enhanced expression of TGF-beta1 in BM-Mp.The defective APC function of macrophages generated in vitro with M-CSF and IL-6 was mainly due to the production of TGF-beta1 by macrophages.

View Article: PubMed Central - PubMed

Affiliation: School of Science Education (Biology), Chungbuk National University, Cheongju, Korea.

ABSTRACT

Background: Macrophages generated in vitro using macrophage-colony stimulating factor (M-CSF) and interleukin (IL)-6 from bone marrow cells (BM-Mp) are defective in antigen presenting cell (APC) function as shown by their ability to induce the proliferation of anti-CD3 mAb-primed syngeneic T cells. However, they do express major histocompatibility (MHC) class I and II molecules, accessory molecules and intracellular adhesion molecules. Here we demonstrate that the defective APC function of macrophages is mainly due to production of TGF-beta1 by BM-Mp.

Methods: Microarray analysis showed that TGF-beta1 was highly expressed in BM-Mp, compared to a macrophage cell line, B6D, which exerted efficient APC function. Production of TGF-beta1 by BM-Mp was confirmed by neutralization experiments of TGF-beta1 as well as by real time-polymerase chain reaction (PCR).

Results: Addition of anti-TGF-beta1 monoclonal antibody to cultures of BM-Mp and anti-CD3 mAb-primed syngeneic T cells efficiently induced the proliferation of syngeneic T cells. Conversely, the APC function of B6D cells was almost completely suppressed by addition of TGF-beta1. Quantitative real time-PCR analysis also confirmed the enhanced expression of TGF-beta1 in BM-Mp.

Conclusion: The defective APC function of macrophages generated in vitro with M-CSF and IL-6 was mainly due to the production of TGF-beta1 by macrophages.

No MeSH data available.


Related in: MedlinePlus

Recovery of APC function of BM-Mp by blocking with anti-TGF-β1 mAb. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with BM-Mp (1×104 cells/well) in the presence of the indicated amounts of anti-TGF-β1 mAb. IgG is an isotype control for anti-TGF-β1 mAb. The results show the mean±S.D. of three independent experiments.
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Figure 4: Recovery of APC function of BM-Mp by blocking with anti-TGF-β1 mAb. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with BM-Mp (1×104 cells/well) in the presence of the indicated amounts of anti-TGF-β1 mAb. IgG is an isotype control for anti-TGF-β1 mAb. The results show the mean±S.D. of three independent experiments.

Mentions: To confirm that TGF-β1 produced from BM-Mp was responsible for the defective APC function of BM-Mp, two experiments, blocking of TGF-β1 with anti-TGF-β1 mAbs and addition of TGF-β1, were performed. As shown in Fig. 4, addition of anti-TGF-β1 mAbs to mixed cultures of BM-Mp and anti-CD3 mAb-primed syngeneic T cells dose-dependently increased the proliferation of T cells. Conversely, addition of TGF-β1 to mixed cultures of B6D cells and anti-CD3 mAb-primed syngeneic T cells dose-dependently inhibited the proliferation of T cells (Fig. 5). These results indicated that TGF-β1 produced from BM-Mp was responsible for the defective APC function of BM-Mp.


Production of TGF-beta1 as a Mechanism for Defective Antigen-presenting Cell Function of Macrophages Generated in vitro with M-CSF.

Lee JK, Lee YR, Lee YH, Kim K, Lee CK - Immune Netw (2009)

Recovery of APC function of BM-Mp by blocking with anti-TGF-β1 mAb. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with BM-Mp (1×104 cells/well) in the presence of the indicated amounts of anti-TGF-β1 mAb. IgG is an isotype control for anti-TGF-β1 mAb. The results show the mean±S.D. of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2803298&req=5

Figure 4: Recovery of APC function of BM-Mp by blocking with anti-TGF-β1 mAb. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with BM-Mp (1×104 cells/well) in the presence of the indicated amounts of anti-TGF-β1 mAb. IgG is an isotype control for anti-TGF-β1 mAb. The results show the mean±S.D. of three independent experiments.
Mentions: To confirm that TGF-β1 produced from BM-Mp was responsible for the defective APC function of BM-Mp, two experiments, blocking of TGF-β1 with anti-TGF-β1 mAbs and addition of TGF-β1, were performed. As shown in Fig. 4, addition of anti-TGF-β1 mAbs to mixed cultures of BM-Mp and anti-CD3 mAb-primed syngeneic T cells dose-dependently increased the proliferation of T cells. Conversely, addition of TGF-β1 to mixed cultures of B6D cells and anti-CD3 mAb-primed syngeneic T cells dose-dependently inhibited the proliferation of T cells (Fig. 5). These results indicated that TGF-β1 produced from BM-Mp was responsible for the defective APC function of BM-Mp.

Bottom Line: Production of TGF-beta1 by BM-Mp was confirmed by neutralization experiments of TGF-beta1 as well as by real time-polymerase chain reaction (PCR).Quantitative real time-PCR analysis also confirmed the enhanced expression of TGF-beta1 in BM-Mp.The defective APC function of macrophages generated in vitro with M-CSF and IL-6 was mainly due to the production of TGF-beta1 by macrophages.

View Article: PubMed Central - PubMed

Affiliation: School of Science Education (Biology), Chungbuk National University, Cheongju, Korea.

ABSTRACT

Background: Macrophages generated in vitro using macrophage-colony stimulating factor (M-CSF) and interleukin (IL)-6 from bone marrow cells (BM-Mp) are defective in antigen presenting cell (APC) function as shown by their ability to induce the proliferation of anti-CD3 mAb-primed syngeneic T cells. However, they do express major histocompatibility (MHC) class I and II molecules, accessory molecules and intracellular adhesion molecules. Here we demonstrate that the defective APC function of macrophages is mainly due to production of TGF-beta1 by BM-Mp.

Methods: Microarray analysis showed that TGF-beta1 was highly expressed in BM-Mp, compared to a macrophage cell line, B6D, which exerted efficient APC function. Production of TGF-beta1 by BM-Mp was confirmed by neutralization experiments of TGF-beta1 as well as by real time-polymerase chain reaction (PCR).

Results: Addition of anti-TGF-beta1 monoclonal antibody to cultures of BM-Mp and anti-CD3 mAb-primed syngeneic T cells efficiently induced the proliferation of syngeneic T cells. Conversely, the APC function of B6D cells was almost completely suppressed by addition of TGF-beta1. Quantitative real time-PCR analysis also confirmed the enhanced expression of TGF-beta1 in BM-Mp.

Conclusion: The defective APC function of macrophages generated in vitro with M-CSF and IL-6 was mainly due to the production of TGF-beta1 by macrophages.

No MeSH data available.


Related in: MedlinePlus