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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.

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Calcium flux responses of leukocytes to MCP-5. Fura-2–loaded  cells were exposed sequentially to the indicated chemokines with their  concentrations in parentheses. Calcium flux is reported as ratio fluorescence of fura-2 in human peripheral blood mononuclear cells (A), murine  eosinophils (B), and murine neutrophils (C). The results shown are representative experiments (n ⩾4 for all cell types).
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Figure 3: Calcium flux responses of leukocytes to MCP-5. Fura-2–loaded cells were exposed sequentially to the indicated chemokines with their concentrations in parentheses. Calcium flux is reported as ratio fluorescence of fura-2 in human peripheral blood mononuclear cells (A), murine eosinophils (B), and murine neutrophils (C). The results shown are representative experiments (n ⩾4 for all cell types).

Mentions: Chemokines induce cell migration and activation by binding to specific G protein–coupled seven transmembrane cell surface receptors on leukocytes. Signaling through these receptors results in a transient rise in intracellular calcium. To determine if MCP-5 induces a calcium flux in responding leukocytes, and thus further examine the leukocyte specificity of MCP-5, purified leukocyte subsets were loaded with the calcium sensitive dye fura-2, and their response to MCP-5 was monitored by fluorimetry. Concordant with the chemotaxis data presented above, MCP-5 induced a dose dependent calcium flux in mononuclear cells (Fig. 3 A), but not eosinophils (even at 50 μg/ml or 5 μM) (Fig. 3 B), or neutrophils (Fig. 3 C). As controls, the purified eosinophils responded appropriately to eotaxin and MIP-1α, but did not respond to MIP-1β (Fig. 3 B) or JE (data not shown), and the purified neutrophils responded appropriately to KC and MIP-1α (Fig. 3 C). The half-maximal effective concentration of the MCP-5 induced mononuclear cell calcium transient was ∼10 ng/ml (1 nM) (data not shown). These results demonstrate that MCP-5 induces a calcium flux in mononuclear cells, but not in eosinophils or neutrophils.


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)

Calcium flux responses of leukocytes to MCP-5. Fura-2–loaded  cells were exposed sequentially to the indicated chemokines with their  concentrations in parentheses. Calcium flux is reported as ratio fluorescence of fura-2 in human peripheral blood mononuclear cells (A), murine  eosinophils (B), and murine neutrophils (C). The results shown are representative experiments (n ⩾4 for all cell types).
© Copyright Policy
Related In: Results  -  Collection

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

Figure 3: Calcium flux responses of leukocytes to MCP-5. Fura-2–loaded cells were exposed sequentially to the indicated chemokines with their concentrations in parentheses. Calcium flux is reported as ratio fluorescence of fura-2 in human peripheral blood mononuclear cells (A), murine eosinophils (B), and murine neutrophils (C). The results shown are representative experiments (n ⩾4 for all cell types).
Mentions: Chemokines induce cell migration and activation by binding to specific G protein–coupled seven transmembrane cell surface receptors on leukocytes. Signaling through these receptors results in a transient rise in intracellular calcium. To determine if MCP-5 induces a calcium flux in responding leukocytes, and thus further examine the leukocyte specificity of MCP-5, purified leukocyte subsets were loaded with the calcium sensitive dye fura-2, and their response to MCP-5 was monitored by fluorimetry. Concordant with the chemotaxis data presented above, MCP-5 induced a dose dependent calcium flux in mononuclear cells (Fig. 3 A), but not eosinophils (even at 50 μg/ml or 5 μM) (Fig. 3 B), or neutrophils (Fig. 3 C). As controls, the purified eosinophils responded appropriately to eotaxin and MIP-1α, but did not respond to MIP-1β (Fig. 3 B) or JE (data not shown), and the purified neutrophils responded appropriately to KC and MIP-1α (Fig. 3 C). The half-maximal effective concentration of the MCP-5 induced mononuclear cell calcium transient was ∼10 ng/ml (1 nM) (data not shown). These results demonstrate that MCP-5 induces a calcium flux in mononuclear cells, but not in eosinophils or neutrophils.

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