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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

Comparison of the APC functions of BM-Mp and B6D cells. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with the indicated number of BM-Mp or B6D cells. Proliferation of T cells was measured by [3H]-thymidine incorporation for the final 8 h of the culture period of 3 days. The results show the mean±S.D. of three independent experiments.
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Figure 3: Comparison of the APC functions of BM-Mp and B6D cells. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with the indicated number of BM-Mp or B6D cells. Proliferation of T cells was measured by [3H]-thymidine incorporation for the final 8 h of the culture period of 3 days. The results show the mean±S.D. of three independent experiments.

Mentions: The APC function of B6D cells and BM-Mp was comparatively studied by testing their ability to induce proliferation of syngeneic anti-CD3 mAb-primed T cells. Consistent with previous observations (4), BM-Mp cells were defective in inducing proliferation of syngeneic anti-CD3 mAb-primed T cells. However, B6D cells efficiently enhanced the proliferation of syngeneic anti-CD3 mAb-primed T cells (Fig. 3). The proliferation-inducing activity of B6D cells was most potent when the ratio of B6D cells and syngeneic anti-CD3 mAb-primed T cells was 1:10.


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)

Comparison of the APC functions of BM-Mp and B6D cells. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with the indicated number of BM-Mp or B6D cells. Proliferation of T cells was measured by [3H]-thymidine incorporation for the final 8 h of the culture period of 3 days. 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 3: Comparison of the APC functions of BM-Mp and B6D cells. Syngeneic anti-CD3 mAb-primed T cells (1×105 cells/well) were cultured with the indicated number of BM-Mp or B6D cells. Proliferation of T cells was measured by [3H]-thymidine incorporation for the final 8 h of the culture period of 3 days. The results show the mean±S.D. of three independent experiments.
Mentions: The APC function of B6D cells and BM-Mp was comparatively studied by testing their ability to induce proliferation of syngeneic anti-CD3 mAb-primed T cells. Consistent with previous observations (4), BM-Mp cells were defective in inducing proliferation of syngeneic anti-CD3 mAb-primed T cells. However, B6D cells efficiently enhanced the proliferation of syngeneic anti-CD3 mAb-primed T cells (Fig. 3). The proliferation-inducing activity of B6D cells was most potent when the ratio of B6D cells and syngeneic anti-CD3 mAb-primed T cells was 1:10.

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