The essentiality of PKCα and PKCβ1 translocation for CD14+monocyte differentiation towards macrophages and dendritic cells, respectively

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Abstract

Human peripheral CD14+monocytes have been known to differentiate into monocyte-derived macrophages (MDMs) or dendritic cells (MoDCs) upon suitable stimulation. However, the key intracellular molecule(s) associated with their differentiation towards specific cell types was(were) not fully understood. This study was designated to determine the association of PKC isoenzymes with the differentiation of CD14+monocytes into MDMs or MoDCs. Purified human peripheral CD14+monocytes were cultured with GM-CSF, or GM-CSF plus IL-4 for 7 days to induce cell differentiation. The phenotypic changes were analyzed by Flow-Cytometry using various specific antibodies to cell type-specific surface markers. The immunological functions of these differentiated cells were determined by measuring the amounts of TNF-α secretion for MDMs, and the capacities of antigen-capturing and bacterial phagocytosis for MoDCs. The translocations of PKC isoenzymes in these cells from cytosol to plasma membrane were examined by Western Blot analysis and Confocal Microscopic observation. The treatment of CD14+monocytes with either GM-CSF or PMA elicited PKCα translocation and consequently induced their differentiation into MDMs. The inclusion of PKCα/ β1 specific inhibitor, Go6976, greatly inhibited the GM-CSF-induced PKCα translocation and dose-dependently reduced the GM-CSF- induced MDM differentiation. On the other hand, the simultaneous pretreatment of CD14+monocytes with Go6976 and PKCβ-specific inhibitor predominantly suppressed the GM-CSF/IL-4-induced generation of MoDCs. Further study demonstrated that GM-CSF/IL-4 selectively induced the translocation of PKCβ1, not PKCα or PKCβ1, in CD14 +monocytes. In conclusion, the cell fate commitment of CD14 +monocytes towards MDMs or MoDCs appears to be steered by the selective activation of PKCα or PKCβ1, respectively.

Original languageEnglish
Pages (from-to)429-441
Number of pages13
JournalJournal of Cellular Biochemistry
Volume102
Issue number2
DOIs
Publication statusPublished - Oct 1 2007

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Macrophages
Granulocyte-Macrophage Colony-Stimulating Factor
Dendritic Cells
Monocytes
Interleukin-4
Isoenzymes
Bacterial Antigens
Flow cytometry
Interleukin-7
Cell membranes
Phagocytosis
Cytosol
Chemical activation
Cell Differentiation
Association reactions
Flow Cytometry
Western Blotting
Cell Membrane
Molecules
Antibodies

Keywords

  • CD14monocytes
  • Dendritic cells
  • Differentiation
  • Macrophages
  • Protein kinase C

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

@article{3abcdde89da34da69c5bccf032aa44d9,
title = "The essentiality of PKCα and PKCβ1 translocation for CD14+monocyte differentiation towards macrophages and dendritic cells, respectively",
abstract = "Human peripheral CD14+monocytes have been known to differentiate into monocyte-derived macrophages (MDMs) or dendritic cells (MoDCs) upon suitable stimulation. However, the key intracellular molecule(s) associated with their differentiation towards specific cell types was(were) not fully understood. This study was designated to determine the association of PKC isoenzymes with the differentiation of CD14+monocytes into MDMs or MoDCs. Purified human peripheral CD14+monocytes were cultured with GM-CSF, or GM-CSF plus IL-4 for 7 days to induce cell differentiation. The phenotypic changes were analyzed by Flow-Cytometry using various specific antibodies to cell type-specific surface markers. The immunological functions of these differentiated cells were determined by measuring the amounts of TNF-α secretion for MDMs, and the capacities of antigen-capturing and bacterial phagocytosis for MoDCs. The translocations of PKC isoenzymes in these cells from cytosol to plasma membrane were examined by Western Blot analysis and Confocal Microscopic observation. The treatment of CD14+monocytes with either GM-CSF or PMA elicited PKCα translocation and consequently induced their differentiation into MDMs. The inclusion of PKCα/ β1 specific inhibitor, Go6976, greatly inhibited the GM-CSF-induced PKCα translocation and dose-dependently reduced the GM-CSF- induced MDM differentiation. On the other hand, the simultaneous pretreatment of CD14+monocytes with Go6976 and PKCβ-specific inhibitor predominantly suppressed the GM-CSF/IL-4-induced generation of MoDCs. Further study demonstrated that GM-CSF/IL-4 selectively induced the translocation of PKCβ1, not PKCα or PKCβ1, in CD14 +monocytes. In conclusion, the cell fate commitment of CD14 +monocytes towards MDMs or MoDCs appears to be steered by the selective activation of PKCα or PKCβ1, respectively.",
keywords = "CD14monocytes, Dendritic cells, Differentiation, Macrophages, Protein kinase C",
author = "Lin, {Yuan Feng} and Lee, {Horng Mo} and Leu, {Sy Jye} and Yu-Hui Tsai",
year = "2007",
month = "10",
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doi = "10.1002/jcb.21305",
language = "English",
volume = "102",
pages = "429--441",
journal = "Journal of Cellular Biochemistry",
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TY - JOUR

T1 - The essentiality of PKCα and PKCβ1 translocation for CD14+monocyte differentiation towards macrophages and dendritic cells, respectively

AU - Lin, Yuan Feng

AU - Lee, Horng Mo

AU - Leu, Sy Jye

AU - Tsai, Yu-Hui

PY - 2007/10/1

Y1 - 2007/10/1

N2 - Human peripheral CD14+monocytes have been known to differentiate into monocyte-derived macrophages (MDMs) or dendritic cells (MoDCs) upon suitable stimulation. However, the key intracellular molecule(s) associated with their differentiation towards specific cell types was(were) not fully understood. This study was designated to determine the association of PKC isoenzymes with the differentiation of CD14+monocytes into MDMs or MoDCs. Purified human peripheral CD14+monocytes were cultured with GM-CSF, or GM-CSF plus IL-4 for 7 days to induce cell differentiation. The phenotypic changes were analyzed by Flow-Cytometry using various specific antibodies to cell type-specific surface markers. The immunological functions of these differentiated cells were determined by measuring the amounts of TNF-α secretion for MDMs, and the capacities of antigen-capturing and bacterial phagocytosis for MoDCs. The translocations of PKC isoenzymes in these cells from cytosol to plasma membrane were examined by Western Blot analysis and Confocal Microscopic observation. The treatment of CD14+monocytes with either GM-CSF or PMA elicited PKCα translocation and consequently induced their differentiation into MDMs. The inclusion of PKCα/ β1 specific inhibitor, Go6976, greatly inhibited the GM-CSF-induced PKCα translocation and dose-dependently reduced the GM-CSF- induced MDM differentiation. On the other hand, the simultaneous pretreatment of CD14+monocytes with Go6976 and PKCβ-specific inhibitor predominantly suppressed the GM-CSF/IL-4-induced generation of MoDCs. Further study demonstrated that GM-CSF/IL-4 selectively induced the translocation of PKCβ1, not PKCα or PKCβ1, in CD14 +monocytes. In conclusion, the cell fate commitment of CD14 +monocytes towards MDMs or MoDCs appears to be steered by the selective activation of PKCα or PKCβ1, respectively.

AB - Human peripheral CD14+monocytes have been known to differentiate into monocyte-derived macrophages (MDMs) or dendritic cells (MoDCs) upon suitable stimulation. However, the key intracellular molecule(s) associated with their differentiation towards specific cell types was(were) not fully understood. This study was designated to determine the association of PKC isoenzymes with the differentiation of CD14+monocytes into MDMs or MoDCs. Purified human peripheral CD14+monocytes were cultured with GM-CSF, or GM-CSF plus IL-4 for 7 days to induce cell differentiation. The phenotypic changes were analyzed by Flow-Cytometry using various specific antibodies to cell type-specific surface markers. The immunological functions of these differentiated cells were determined by measuring the amounts of TNF-α secretion for MDMs, and the capacities of antigen-capturing and bacterial phagocytosis for MoDCs. The translocations of PKC isoenzymes in these cells from cytosol to plasma membrane were examined by Western Blot analysis and Confocal Microscopic observation. The treatment of CD14+monocytes with either GM-CSF or PMA elicited PKCα translocation and consequently induced their differentiation into MDMs. The inclusion of PKCα/ β1 specific inhibitor, Go6976, greatly inhibited the GM-CSF-induced PKCα translocation and dose-dependently reduced the GM-CSF- induced MDM differentiation. On the other hand, the simultaneous pretreatment of CD14+monocytes with Go6976 and PKCβ-specific inhibitor predominantly suppressed the GM-CSF/IL-4-induced generation of MoDCs. Further study demonstrated that GM-CSF/IL-4 selectively induced the translocation of PKCβ1, not PKCα or PKCβ1, in CD14 +monocytes. In conclusion, the cell fate commitment of CD14 +monocytes towards MDMs or MoDCs appears to be steered by the selective activation of PKCα or PKCβ1, respectively.

KW - CD14monocytes

KW - Dendritic cells

KW - Differentiation

KW - Macrophages

KW - Protein kinase C

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