Limits...
Murine monocyte chemoattractant protein (MCP)-5: a novel CC chemokine that is a structural and functional homologue of human MCP-1.

Sarafi MN, Garcia-Zepeda EA, MacLean JA, Charo IF, Luster AD - J. Exp. Med. (1997)

Bottom Line: Consistent with these results, MCP-5 induced a calcium flux in human embryonic kidney (HEK)-293 cells transfected with human and murine CCR2, a CC chemokine receptor expressed on monocytes.MCP-5 did not induce a calcium flux in HEK-293 cells transfected with CCR1, CCR3, or CCR5.These data indicate that MCP-5 is a novel and potent monocyte active chemokine that is involved in allergic inflammation and the host response to pathogens.

View Article: PubMed Central - PubMed

Affiliation: Infectious Disease Unit, Massachusetts General Hospital, Boston, USA.

ABSTRACT
The chemokines are a large family of cytokines that control the recruitment of leukocytes in immune and inflammatory responses. We describe the isolation of a novel murine CC chemokine that, based on its biological and structural features, we have named monocyte chemoattractant protein (MCP)-5. MCP-5 mapped to the CC chemokine cluster on mouse chromosome 11 and was most closely related to human MCP-1 in structure (66% amino acid identity). Purified recombinant MCP-5 protein was a potent chemoattractant for peripheral blood monocytes, was only weakly active on eosinophils at high doses, and was inactive on neutrophils. MCP-5 induced a calcium flux in peripheral blood mononuclear cells, but not in purified murine eosinophils or neutrophils. Consistent with these results, MCP-5 induced a calcium flux in human embryonic kidney (HEK)-293 cells transfected with human and murine CCR2, a CC chemokine receptor expressed on monocytes. MCP-5 did not induce a calcium flux in HEK-293 cells transfected with CCR1, CCR3, or CCR5. Constitutive expression of MCP-5 mRNA was detected predominantly in lymph nodes, and its expression was markedly induced in macrophages activated in vitro and in vivo. Moreover, MCP-5 expression was up-regulated in the lungs of mice following aerosolized antigen challenge of sensitized mice, and during the host response to infection with Nippostrongylus brasiliensis. These data indicate that MCP-5 is a novel and potent monocyte active chemokine that is involved in allergic inflammation and the host response to pathogens.

Show MeSH

Related in: MedlinePlus

MCP-5 mRNA expression in murine leukocytes and endothelial cells. (A) Northern analysis of total RNA isolated from eosinophils  purified from the spleens of CD5-IL5 transgenic mice (Eos), bone marrow–derived mast cells either untreated (Ctl) or stimulated with IgE antiTNP and TNP-BSA (IgE), or Con A, resident peritoneal macrophages  (RMφ) (93% mononuclear, 7% granulocytes), elicited peritoneal macrophages (EMφ) (76% mononuclear, 24% neutrophils), and elicited peritoneal neutrophils (95% neutrophils, 5% mononuclear). Blots were hybridized with MCP-5, JE, and rpL32 cDNA probes and exposed for 14 d,  3 d, and 18 h, respectively. (B) Northern analysis of total RNA isolated  from SVEC cells treated for 6 and 18 h with IFNγ (200 U/ml), IL-4 (10  ng/ml), and IL-1β (5 ng/ml) and 10 μg of total RNA isolated from  RAW 264.7 macrophage cell line, either untreated or treated for 18 h with  IFNγ (200 U/ml) or the indicated concentrations of LPS. Blots were hybridized sequentially with MCP-5, JE, and rpL32 cDNA probes and exposed for 7 d, 3 d, and 18 h, respectively.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196097&req=5

Figure 7: MCP-5 mRNA expression in murine leukocytes and endothelial cells. (A) Northern analysis of total RNA isolated from eosinophils purified from the spleens of CD5-IL5 transgenic mice (Eos), bone marrow–derived mast cells either untreated (Ctl) or stimulated with IgE antiTNP and TNP-BSA (IgE), or Con A, resident peritoneal macrophages (RMφ) (93% mononuclear, 7% granulocytes), elicited peritoneal macrophages (EMφ) (76% mononuclear, 24% neutrophils), and elicited peritoneal neutrophils (95% neutrophils, 5% mononuclear). Blots were hybridized with MCP-5, JE, and rpL32 cDNA probes and exposed for 14 d, 3 d, and 18 h, respectively. (B) Northern analysis of total RNA isolated from SVEC cells treated for 6 and 18 h with IFNγ (200 U/ml), IL-4 (10 ng/ml), and IL-1β (5 ng/ml) and 10 μg of total RNA isolated from RAW 264.7 macrophage cell line, either untreated or treated for 18 h with IFNγ (200 U/ml) or the indicated concentrations of LPS. Blots were hybridized sequentially with MCP-5, JE, and rpL32 cDNA probes and exposed for 7 d, 3 d, and 18 h, respectively.

Mentions: The expression of MCP-5 was examined in purified leukocyte subsets isolated from normal mice (Fig. 7 A). MCP-5 expression was not seen in eosinophils, bone marrow–derived mast cells, or resident peritoneal macrophages. Stimulation of these mast cells with either IgE or Con A did not induce MCP-5 expression. Con A upregulated the expression of JE (Fig. 7 A), and both treatments induced the expression of murine CCR1 (21). Activated macrophages elicited into the peritoneal cavity by thioglycollate expressed significant levels of MCP-5 mRNA. Neutrophils elicited into the peritoneum by treatment with sodium casein also expressed MCP-5 mRNA; however, expression from contaminating elicited macrophages cannot be excluded.


Murine monocyte chemoattractant protein (MCP)-5: a novel CC chemokine that is a structural and functional homologue of human MCP-1.

Sarafi MN, Garcia-Zepeda EA, MacLean JA, Charo IF, Luster AD - J. Exp. Med. (1997)

MCP-5 mRNA expression in murine leukocytes and endothelial cells. (A) Northern analysis of total RNA isolated from eosinophils  purified from the spleens of CD5-IL5 transgenic mice (Eos), bone marrow–derived mast cells either untreated (Ctl) or stimulated with IgE antiTNP and TNP-BSA (IgE), or Con A, resident peritoneal macrophages  (RMφ) (93% mononuclear, 7% granulocytes), elicited peritoneal macrophages (EMφ) (76% mononuclear, 24% neutrophils), and elicited peritoneal neutrophils (95% neutrophils, 5% mononuclear). Blots were hybridized with MCP-5, JE, and rpL32 cDNA probes and exposed for 14 d,  3 d, and 18 h, respectively. (B) Northern analysis of total RNA isolated  from SVEC cells treated for 6 and 18 h with IFNγ (200 U/ml), IL-4 (10  ng/ml), and IL-1β (5 ng/ml) and 10 μg of total RNA isolated from  RAW 264.7 macrophage cell line, either untreated or treated for 18 h with  IFNγ (200 U/ml) or the indicated concentrations of LPS. Blots were hybridized sequentially with MCP-5, JE, and rpL32 cDNA probes and exposed for 7 d, 3 d, and 18 h, respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196097&req=5

Figure 7: MCP-5 mRNA expression in murine leukocytes and endothelial cells. (A) Northern analysis of total RNA isolated from eosinophils purified from the spleens of CD5-IL5 transgenic mice (Eos), bone marrow–derived mast cells either untreated (Ctl) or stimulated with IgE antiTNP and TNP-BSA (IgE), or Con A, resident peritoneal macrophages (RMφ) (93% mononuclear, 7% granulocytes), elicited peritoneal macrophages (EMφ) (76% mononuclear, 24% neutrophils), and elicited peritoneal neutrophils (95% neutrophils, 5% mononuclear). Blots were hybridized with MCP-5, JE, and rpL32 cDNA probes and exposed for 14 d, 3 d, and 18 h, respectively. (B) Northern analysis of total RNA isolated from SVEC cells treated for 6 and 18 h with IFNγ (200 U/ml), IL-4 (10 ng/ml), and IL-1β (5 ng/ml) and 10 μg of total RNA isolated from RAW 264.7 macrophage cell line, either untreated or treated for 18 h with IFNγ (200 U/ml) or the indicated concentrations of LPS. Blots were hybridized sequentially with MCP-5, JE, and rpL32 cDNA probes and exposed for 7 d, 3 d, and 18 h, respectively.
Mentions: The expression of MCP-5 was examined in purified leukocyte subsets isolated from normal mice (Fig. 7 A). MCP-5 expression was not seen in eosinophils, bone marrow–derived mast cells, or resident peritoneal macrophages. Stimulation of these mast cells with either IgE or Con A did not induce MCP-5 expression. Con A upregulated the expression of JE (Fig. 7 A), and both treatments induced the expression of murine CCR1 (21). Activated macrophages elicited into the peritoneal cavity by thioglycollate expressed significant levels of MCP-5 mRNA. Neutrophils elicited into the peritoneum by treatment with sodium casein also expressed MCP-5 mRNA; however, expression from contaminating elicited macrophages cannot be excluded.

Bottom Line: Consistent with these results, MCP-5 induced a calcium flux in human embryonic kidney (HEK)-293 cells transfected with human and murine CCR2, a CC chemokine receptor expressed on monocytes.MCP-5 did not induce a calcium flux in HEK-293 cells transfected with CCR1, CCR3, or CCR5.These data indicate that MCP-5 is a novel and potent monocyte active chemokine that is involved in allergic inflammation and the host response to pathogens.

View Article: PubMed Central - PubMed

Affiliation: Infectious Disease Unit, Massachusetts General Hospital, Boston, USA.

ABSTRACT
The chemokines are a large family of cytokines that control the recruitment of leukocytes in immune and inflammatory responses. We describe the isolation of a novel murine CC chemokine that, based on its biological and structural features, we have named monocyte chemoattractant protein (MCP)-5. MCP-5 mapped to the CC chemokine cluster on mouse chromosome 11 and was most closely related to human MCP-1 in structure (66% amino acid identity). Purified recombinant MCP-5 protein was a potent chemoattractant for peripheral blood monocytes, was only weakly active on eosinophils at high doses, and was inactive on neutrophils. MCP-5 induced a calcium flux in peripheral blood mononuclear cells, but not in purified murine eosinophils or neutrophils. Consistent with these results, MCP-5 induced a calcium flux in human embryonic kidney (HEK)-293 cells transfected with human and murine CCR2, a CC chemokine receptor expressed on monocytes. MCP-5 did not induce a calcium flux in HEK-293 cells transfected with CCR1, CCR3, or CCR5. Constitutive expression of MCP-5 mRNA was detected predominantly in lymph nodes, and its expression was markedly induced in macrophages activated in vitro and in vivo. Moreover, MCP-5 expression was up-regulated in the lungs of mice following aerosolized antigen challenge of sensitized mice, and during the host response to infection with Nippostrongylus brasiliensis. These data indicate that MCP-5 is a novel and potent monocyte active chemokine that is involved in allergic inflammation and the host response to pathogens.

Show MeSH
Related in: MedlinePlus