Comparison of biological responses of human monocytes and THP-1 cells to chemokines of the intercrine-<i>β</i> family

Vaddi, Krishna; Newton, Robert

doi:10.1002/jlb.55.6.756

Cited by 56 publications

(35 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The MIP-1␣ effect on THP-1 cells was dosedependent, and at 1 nM it produced an approximately 50 -60% increase in extracellular acidification. These kinetics are quite similar to those reported for chemokines in human monocytes and in transfected cell lines (24,25). Pretreatment of THP-1 cells with increasing concentrations of compound 1 dose-responsively inhibited the ability of MIP-1␣ to induce changes in the metabolic activity of the cells (Fig.…”

Section: Resultssupporting

confidence: 83%

Identification and Characterization of Small Molecule Functional Antagonists of the CCR1 Chemokine Receptor

Hesselgesser¹,

Ng²,

Liang³

et al. 1998

Journal of Biological Chemistry

130

View full text Add to dashboard Cite

The CC chemokines macrophage inflammatory protein-1␣ (MIP-1␣) and RANTES (regulated on activation normal T cell expressed) have been implicated in rheumatoid arthritis and multiple sclerosis. Since their effects are mediated through the CCR1 chemokine receptor, we set up a small molecule CCR1 antagonist program to search for inhibitors. Through high capacity screening we discovered a number of 4-hydroxypiperidine compounds with CCR1 antagonist activity and report their synthesis and in vitro pharmacology here. Scatchard analysis of the competition binding data revealed that the compounds had K i values ranging from 40 to 4000 nM. The pharmacological profile of the most potent member of this series, compound 1 (2-2-diphenyl-5-(4-chlorophenyl)piperidin-lyl)valeronitrite), was further evaluated. The directed migration of select populations of leukocytes from the circulation to sites of inflammation is an integral part of the immune response. The chemokines are a diverse group of proteins that play an important role in this process (1). They are classified into two major groups, CXC and CC, based on the position of the first two of their four invariant cysteines (2). Each of the chemokines recognizes and induces the chemotaxis of a particular subset of leukocytes. For example, the CXC chemokines, like IL-8 1 and melonoma growth stimulatory activity, mainly chemoattract and activate neutrophils, in contrast to the CC chemokines, like RANTES and monocyte chemoattractant protein, which preferentially attract T lymphocytes and monocytes and induce their activation by producing changes in cellular morphology, transient increases in cellular calcium concentration, and the up-regulation of surface adhesion proteins.The chemokines produce their biologic effects by interacting with specific receptors on the cell surface of their target cells (3). To date, 14 different chemokine receptors including eight CC chemokine receptors have been identified by cloning (4, 5). All of these receptors are characterized by a heptahelical structure and belong to a superfamily of serpentine receptors that are coupled to guanine nucleotide-binding proteins (G-proteins) (6).Occasionally the immune system can turn upon its host, giving rise to chronic inflammation and disease. Given their important role in this process, chemokines have been implicated in the pathophysiology of autoimmune diseases like multiple sclerosis and rheumatoid arthritis. For example a recent study by Karpus et al. (7) provides strong in vivo concept validation for a role of MIP-1␣ in a mouse experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis. These investigators were able to show that antibodies to MIP-1␣ prevented the development of both acute and relapsing paralytic disease as well as infiltration of mononuclear cells into the central nervous system. Treatment with MIP-1␣ antibody was also able to ameliorate the severity of ongoing clinical disease. These results led the authors to conclude that MIP-1␣ plays an important role in this T-cell me...

show abstract

Section: Resultssupporting

confidence: 83%

Identification and Characterization of Small Molecule Functional Antagonists of the CCR1 Chemokine Receptor

Hesselgesser¹,

Ng²,

Liang³

et al. 1998

Journal of Biological Chemistry

130

View full text Add to dashboard Cite

show abstract

“…The EC 50 for Ca 2ϩ flux in the Jurkat CCR2B transfectants ranged from approximately 10 -40 nM depending on the level of surface receptor expression, whereas an EC 50 of about 3.4 nM was obtained for Ca 2ϩ flux in CCR2B HEK-293 transfectants that express high levels of CCR2B (17). Peripheral blood monocytes mobilize Ca 2ϩ in response to low nanomolar concentrations of MCP-1 (27)(28)(29), whereas concentrations as high as 10 nM MCP-1 did not mobilize Ca 2ϩ in normal T cells from peripheral blood (27). Therefore, receptor number as well as differences in the receptor coupling between cell types may contribute to differences in functional activity between peripheral T cells and monocytes as well as between CCR2 expressed in Jurkat and HEK-293 cells.…”

Section: Discussionmentioning

confidence: 99%

Functional Differences Between Monocyte Chemotactic Protein-1 Receptor A and Monocyte Chemotactic Protein-1 Receptor B Expressed in a Jurkat T Cell

Sanders

Crean

Boxer

et al. 2000

The Journal of Immunology

View full text Add to dashboard Cite

The monocyte chemotactic protein-1 (MCP-1) receptor (MCP-1R) is expressed on monocytes, a subpopulation of memory T lymphocytes, and basophils. Two alternatively spliced forms of MCP-1R, CCR2A and CCR2B, exist and differ only in their carboxyl-terminal tails. To determine whether CCR2A and CCR2B receptors function similarly, Jurkat T cells were stably transfected with plasmids encoding the human CCR2A or CCR2B gene. Nanomolar concentrations of MCP-1 induced chemotaxis in the CCR2B transfectants that express high, intermediate, and low levels of MCP-1R. Peak chemotactic activity was shifted to the right as receptor number decreased. Five-fold more MCP-1 was required to initiate chemotaxis of the CCR2A low transfectant, but the peak of chemotaxis was similar for the CCR2A and CCR2B transfectants expressing similar numbers of receptors. MCP-1-induced chemotaxis was sensitive to pertussis toxin, implying that both CCR2A and CCR2B are Giα protein coupled. MCP-1 induced a transient Ca2+ flux in the CCR2B transfectant that was partially sensitive to pertussis toxin. In contrast, MCP-1 did not induce Ca2+ flux in the CCR2A transfectant. Since MCP-1 can stimulate chemotaxis of the CCR2A transfectant without inducing Ca2+ mobilization, Ca2+ flux may not be required for MCP-1-induced chemotaxis in the Jurkat transfectants. These results indicate that functional differences exist between the CCR2A and CCR2B transfectants that can be attributed solely to differences in the carboxyl-terminal tail.

show abstract

“…Other roles for oligomerization are certainly possible and may be relevant to additional processes triggered by chemokine binding such as lysozomal enzyme release and generation of toxic products from the respiratory burst (69,70). Alternatively, oligomerization may be involved in as yet unidentified functions unrelated to chemotaxis.…”

Section: Disulfide Cross-linked Wt*(c77) and [1ϩ9 -76](c77) Bind And mentioning

confidence: 99%

Monomeric Monocyte Chemoattractant Protein-1 (MCP-1) Binds and Activates the MCP-1 Receptor CCR2B

Paavola¹,

Hemmerich²,

Grünberger³

et al. 1998

Journal of Biological Chemistry

184

223

View full text Add to dashboard Cite

To address the role of dimerization in the function of the monocyte chemoattractant protein-1, MCP-1, we mutated residues that comprise the core of the dimerization interface and characterized the ability of these mutants to dimerize and to bind and activate the MCP-1 receptor, CCR2b. One mutant, P8A*, does not dimerize. However, it has wild type binding affinity, stimulates chemotaxis, inhibits adenylate cyclase, and stimulates calcium influx with wild type potency and efficacy. These data suggest that MCP-1 binds and activates its receptor as a monomer. In contrast, Y13A*, another monomeric mutant, has a 100-fold weaker binding affinity, is a much less potent inhibitor of adenylate cyclase and stimulator of calcium influx, and is unable to stimulate chemotaxis. Thus Tyr 13 may make important contacts with the receptor that are required for high affinity binding and signal transduction. We also explored whether a mutant, Chemokines are small secreted proteins that function as intercellular messengers to control migration and activation of specific subsets of leukocytes (1-3). This process is mediated by the interaction of chemokines with seven transmembrane Gprotein-coupled receptors on the surface of target cells. Interest in these proteins was first stimulated by the observation of elevated levels in a number of inflammatory diseases (4, 5) including rheumatoid arthritis (6, 7), arteriosclerosis (8, 9), and asthma (10). Although it is not clear whether excessive production is the cause or consequence of these diseases, the demonstration that neutralizing antibodies (11-13) reduced symptoms in a number of animal models generated optimism that receptor antagonists may have therapeutic benefit (14). Recently, it has been shown that certain chemokine receptors also serve as obligate coreceptors for entry of the human immunodeficiency virus into CD4ϩ cells (15)(16)(17) and that viral replication can be inhibited by the ligands of the coreceptors (18 -21). Thus a wide range of clinically important diseases are associated with chemokines and their receptors, motivating many studies to understand the molecular details of chemokine function.Chemokines have been classified into two major families based on their pattern of cysteine residues, their chromosomal location, and their cell specificities (22). ␣-Chemokines such as IL-8 1 have a conserved CXC cysteine motif and act predominantly on neutrophils, whereas ␤-chemokines have a CC signature and attract monocytes and T-cells. The recently discovered chemokines lymphotactin (23) and fractalkine/neurotactin (24,25) are characterized by C and CX 3 C motifs, respectively, and chemoattract T-cells and NK cells. Mutagenesis studies, particularly of ␣-and ␤-chemokines, have provided some insight into the structural determinants of receptor binding and the specificity of these proteins, but many details have yet to emerge.Considerable effort has been devoted to characterizing the stochiometry of chemokine-receptor complexes because most chemokines oligomerize to an extent tha...

show abstract

Comparison of biological responses of human monocytes and THP-1 cells to chemokines of the intercrine-β family

Cited by 56 publications

References 22 publications

Identification and Characterization of Small Molecule Functional Antagonists of the CCR1 Chemokine Receptor

Identification and Characterization of Small Molecule Functional Antagonists of the CCR1 Chemokine Receptor

Functional Differences Between Monocyte Chemotactic Protein-1 Receptor A and Monocyte Chemotactic Protein-1 Receptor B Expressed in a Jurkat T Cell

Monomeric Monocyte Chemoattractant Protein-1 (MCP-1) Binds and Activates the MCP-1 Receptor CCR2B

Contact Info

Product

Resources

About