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Regulation of chemokine receptor expression in dendritic cells
0 INsTITUT PA~TEU~ELSEVIER Paris 1998
Regulation A. Mantovani
Res. Immunol. 1998, 149, 639641
of chemokine receptor expression in dendritic cells (‘jc2), P. Allavena
(‘), A. Vecchi
(‘) and S. Sozzani
(I)
(I) Istituto di Ricerche Fannacologiche “Mario Negri”, Via Eritrea 62, 20157 Milan (Italy), (” Dept. Biotec hnology, Section of General Pathology, Univ. of Brescia, Brescia (Italy)
Introduction Chemokines are a superfamily of small proteins which play a crucial role in immune and inflammatory reactions and in viral infection [l-3]. Most chemokines cause chemotactic migration of leukocytes, but these molecules also affect angiogenesis, collagen production and the proliferation of haemato poietic precursors. Based on a cysteine motif, CXC, CC, C and CX3C families have been identified. The chemokine scaffold consists of an N-terminal loop connected via Cys bonds to the more structured core of the molecule (3 P-sheets) with a C-terminal a helix. About 50 human chemokines have been identified. Chemokines interact with seven transmembrane domain, G-protein-coupled receptors. Eight CC (CCRl through 8), 5 CXC (CXCRl through 5) and 1 CX3C (CX3CRl) receptor have been identified. Receptor expression is a crucial determinant of the spectrum of action of chemokines. For instance, recent results indicate that polarized Thl and Th2 populations show differential receptor expression and responsiveness to chemokines [4-61. Emerging evidence shows that regulation of receptor expression during activation or deactivation of monocytes is as important as regulation of chemokine production for tuning the chemokine system [7, 81. Response
of dendritic
cells to chemokines
Dendritic cell (DC) precursors originate in the bone marrow and subsequently migrate into peripheral tissues and primary lymphoid organs, where they efficiently take up and process soluble antigens. After capture of antigen in tissues, DCs Received July 7, 1998.
and
migrate via the afferent lymphatics to draining lymph nodes or via blood to spleen, where they stimulate T cells [9, lo]. As the molecules controlling these events are unknown, we investigated which chemotactic factors could attract this cell population and regulate their trafficking in tissues [ 1 I]. For these experiments, DCs were obtained from peripheral blood precursor cells cultured in the presence of GM-CSF and IL4 or IL13 [12, 131. These cells were CDla+, MHC class II++, CD14-, CD3- and CD20- and behaved as classical immature DCs, active in eliciting the proliferation of naive T lymphocytes and showing a strong ability to take up FITC-dextran [ 131. In a microwell chemotaxis assay, these cells migrated in response to a selected pattern of CC chemokines, such as MCP-3, RANTES and MIPla and to two prototypic chemotactic factors, fMLP and C5a [ 1 l] (table II). MCP- 1 and MCP-2, two other CC chemokines and all of the C-X-C chemokines tested (IL8, IP-10 and GROP) were inactive. Peak active concentrations and the percentage of input DCs migrating in response to CC chemokines were comparable to those observed with monocytes. MCP-3, MIP-la and RANTES showed a complex pattern of cross-desensitization suggesting the presence of multiple promiscuous receptors on these cells. In northern blot studies, it was found that DCs express the CCRl, CCR2 and CCRS, as well as CXCR2 and CXCR4. DCs respond to SDFI, but not to IL8 and MCP-I in vitro in spite of expressing CCR2 and CXCR2 mRNA. Transgenic mice, in which MCP- 1 is constitutively expressed in the epidermis under the control of the human keratin 14 promoter, respond to a contact
A. MANTOVANI
640
hypersensitivity challenge with an increased lichenoid infiltration of monocytes, T lymphocytes and CD45+, I-A+ cells that assumed a dendritic morphology in situ [ 141.We speculatethat MCP- 1 may attract monocytes to the skin, where thesecells find an environment (e.g. GM-CSF) conducive to DC differentiation.
Differential regulation during maturation
In the context of our interest in chemokines and DC, we recently transfected tumour cells with MCP-3 [ 161. After MCP-3 gene transfer, P8 15 mastocytoma cells grew, but underwent rejection. MCP-3-elicited rejection was associated with resistance to subsequent challenge with parental cells. MCP-3-elicited rejection was associated with profound alterations of leukocyte infiltration. TAMS were already present in large numbers, but T cells, eosinophils and neutrophils increased in tumour tissues after MCP-3 gene transfer. DC (e.g. Dec205+, high MHC class II+ cells) did not increase substantially in the tumour mass. However, in peritumoral tissues, DCs accumulated in perivascular areas. In contrast to their behavior, in immunocompetent mice, MCP-3-transfected tumour cells grew normally in nude mice. Increased accumulation of macrophages and PMN was also evident in nude mice. Antibodies against CD4, CD8 and IFNy, but not against IL4, inhibited rejection of MCP-3 transfected P815 cells. An anti-PMN mAb caused only a delay in MCP-3elicited tumour rejection. Thus, MCP-3 gene transfer elicits tumour rejection by activating type I T-cell-dependent immunity. It is tempting to speculate that altered trafficking of antigen-presenting cells, which express receptors and respond to MCP-3, together with recruitment of activated T cells, underlies activation of specific immunity by MCP-3-transfected cells.
= =
dendritic effective
= =
granulocyte-macrophage interferon.
Key-words: Dendritic cell, Chemokine; Receptor expression, Review. References
PI Baggiolini, M., Dewald, B. & Moser, B. (1997), Human chemokines:An update.Annu. Rev. Immu-
nol., 15,675705.
[21 Hedrick, J.A. & Zlotnik, A. (1996), Chemokinesand lymphocyte biology. Curr. Opin. Immunol., 8, 343347. r31 Rollins, B.J. (1997), Chemokines.Blood, 90, 909928. [41 Bonecchi, R., Bianchi, G., Bordignon, P.P., ef al.
(1998),Differentialexpressionof chemokinereceptors and chemotactic responsiveness of type 1 T helper cells(Thls) andTh2s.J. Exp. Med., 187, 129-134. El Sallusto, F., Mackay, C.R. & Lanzavecchia, A. (1997), Selective expressionof the eotaxin receptor CCR3 by human T helper 2 cells. Science, 277, 2005-2007.
[61 Loetscher, P., Uguccioni, M., Bordoli, L., et al. (1998), CCRS is characteristicof Thl lymphocytes. Nature, 391, 344-345. [71 Sica, A., Saccani,A., Borsatti,A., et al. (1997), Bac-
terial lipopolysacchariderapidly inhibits expression
1
cell. dose. colony-stimulating
factor.
of receptor expression
We have recently investigated how signals which induce maturation of dendritic cells affect their migration [ 17, 181.These studies were based on parallel efforts in monocytes [7, 8, 191. Maturation of DC by CD40 ligation, or culture in the presence of inflammatory agonists, such as bacterial lipopolysaccharide, IL1 and TNF, induces downregulation of the two main CC chemokine receptors expressed by these cells, CCRl and CCRS, and abrogates the chemotactic responseto their ligands, MIP- 1CI,MIPlp, RANTES and MCP-3. Inhibition of chemotaxis was rapid (< 1 h), and included the unrelated agent fMLP. Concomitantly, in the same experimental conditions, the expression of CCR7 and the migration to its ligand ELC/MIP-3/3, a chemokine expressed in lymphoid organs, was strongly upregulated, though with slower kinetics (24-48 h). Rapid inhibition of responsiveness to chemoattractants present at sites of inflammation and immune reaction may be permissive for leaving peripheral tissues. Conversely, the slower acquisition of responsiveness to ELC/MIP-3P may guide subsequent localization of DCs in lymphoid organs.
Macrophage-derived chemokine (MDC) is a recently identified CC chemokine, which, unlike related molecules, is located on chromosome 16 and has a rather distant relationship with other family members [15]. We recently found that MDC has an ED,, lOO-fold lower for activated NK cells and DC than for monocytes [ 151. Moreover, it is expressed by DC in addition to macrophages. We speculate that molecules such as SDF-1 and MDC may contribute to directing the “normal” trafficking of DC in the absence of antigen or inflammation, whereas inducible chemokines may underlie the dramatic changes in route and trafficking after exposure to antigenic or inflammatory stimulation.
DC ED GMCSF IFN
ET AL.
IL MDC PMN TAM
= = = =
interleukin. macrophage-derived polymorphonuclear tumour-associated
chemokine. (cell). macrophage.
CHEMOKINE
[8]
[9] [lo] [l l]
[12]
[ 131
[ 141
RECEPTOR
EXPRESSION
of C-C chemokine receptors in human monocytes. J. Exp. Med., 185, 969-974. Sozzani, S., Ghezzi, S., Iannolo, G. et al. (1998), Interleukin-10 increases CCRS expression and HIV infection in human monocytes. J. Exp. Med., 187, 439-444. Caux, C., Liu, Y.J. & Banchereau, J. (1995), Recent advances in the study of dendritic cells and follicular dendritic cells. Immunol. Today, 16, 2-4. Bancherau, J. & Steinman, R.M. (1998), Dendritic cells and the control of immunity. Nature, 392, 245 252. Sozzani, S., Sallusto, F., Luini, W. et al. (1995), Migration of dendritic cells in response to formyl peptides, C5a and a distinct set of chemokines. J. Immunol., 155, 3292-3295. Sallusto, F. & Lanzavecchia, A. (1994), Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/ macrophage colony-stimulating factor plus interleukin-4 and downregulated by tumor necrosis factor-alpha. J. Exp. Med., 179, 1109-1118. Piemonti, L., Bemasconi, S., Luini, W. et al. (1995), IL-13 supports differentiation of dendritic cells from circulating precursors in concert with GM-CSF. Eur. Cytokine Netw., 6, 245-252. Nakamura, K., Williams, I.R. & Kupper, T.S. (1995),
[15]
[16]
[ 171
[18]
[19]
IN DENDRITIC
CELLS
Keratinocyte-derived monocyte chemoattractant protein 1 (MCP-1): Analysis in a transgenic model demonstrates MCP-1 can recruit dendritic and langerhans cells to skin. J. Invest. Dermatol., 105, 635-643. Godiska, R., Chantry, D., Raport, C.J. etal. (1997), Human macrophage derived chemokine (MDC) a novel chemoattractant for monocytes, monocyte derived dendritic cells, and natural killer cells. J. Exp. Med., 185, 1595-1604. Fioretti, F., Fradelizi, D., Stoppacciaro, A. et al. (1998). Reduced tumorigenicity and augmented leukocyte infiltration after MCP-3 gene transfer: perivascular accumulation of dendritic cells in peritumoral tissue and neutrophil recruitment within the tumor. J. Zmmunol. (in press). Sozzani, S., Allavena, P., D’Amico, G. et al. (1998), Differential regulation of chemokine receptors during dendritic cell maturation: a model for their trafficking properties. J. Immunol. (in press). D’Amico, G., Bianchi, G., Bernasconi, S. et al. (1998), Adhesion, transendothelial migration and reverse transmigration of in vitro cultured dendritic cells. Blood (in press). Penton-Rol, G., Polentarutti, N., Luini, W. et al. (1998), Selective inhibition of expression of the chemokine receptor CCR2 in human monocytes by IFN-y. J. Immunol., 160, 3869-3873.
Regulation A. Mantovani
Res. Immunol. 1998, 149, 639641
of chemokine receptor expression in dendritic cells (‘jc2), P. Allavena
(‘), A. Vecchi
(‘) and S. Sozzani
(I)
(I) Istituto di Ricerche Fannacologiche “Mario Negri”, Via Eritrea 62, 20157 Milan (Italy), (” Dept. Biotec hnology, Section of General Pathology, Univ. of Brescia, Brescia (Italy)
Introduction Chemokines are a superfamily of small proteins which play a crucial role in immune and inflammatory reactions and in viral infection [l-3]. Most chemokines cause chemotactic migration of leukocytes, but these molecules also affect angiogenesis, collagen production and the proliferation of haemato poietic precursors. Based on a cysteine motif, CXC, CC, C and CX3C families have been identified. The chemokine scaffold consists of an N-terminal loop connected via Cys bonds to the more structured core of the molecule (3 P-sheets) with a C-terminal a helix. About 50 human chemokines have been identified. Chemokines interact with seven transmembrane domain, G-protein-coupled receptors. Eight CC (CCRl through 8), 5 CXC (CXCRl through 5) and 1 CX3C (CX3CRl) receptor have been identified. Receptor expression is a crucial determinant of the spectrum of action of chemokines. For instance, recent results indicate that polarized Thl and Th2 populations show differential receptor expression and responsiveness to chemokines [4-61. Emerging evidence shows that regulation of receptor expression during activation or deactivation of monocytes is as important as regulation of chemokine production for tuning the chemokine system [7, 81. Response
of dendritic
cells to chemokines
Dendritic cell (DC) precursors originate in the bone marrow and subsequently migrate into peripheral tissues and primary lymphoid organs, where they efficiently take up and process soluble antigens. After capture of antigen in tissues, DCs Received July 7, 1998.
and
migrate via the afferent lymphatics to draining lymph nodes or via blood to spleen, where they stimulate T cells [9, lo]. As the molecules controlling these events are unknown, we investigated which chemotactic factors could attract this cell population and regulate their trafficking in tissues [ 1 I]. For these experiments, DCs were obtained from peripheral blood precursor cells cultured in the presence of GM-CSF and IL4 or IL13 [12, 131. These cells were CDla+, MHC class II++, CD14-, CD3- and CD20- and behaved as classical immature DCs, active in eliciting the proliferation of naive T lymphocytes and showing a strong ability to take up FITC-dextran [ 131. In a microwell chemotaxis assay, these cells migrated in response to a selected pattern of CC chemokines, such as MCP-3, RANTES and MIPla and to two prototypic chemotactic factors, fMLP and C5a [ 1 l] (table II). MCP- 1 and MCP-2, two other CC chemokines and all of the C-X-C chemokines tested (IL8, IP-10 and GROP) were inactive. Peak active concentrations and the percentage of input DCs migrating in response to CC chemokines were comparable to those observed with monocytes. MCP-3, MIP-la and RANTES showed a complex pattern of cross-desensitization suggesting the presence of multiple promiscuous receptors on these cells. In northern blot studies, it was found that DCs express the CCRl, CCR2 and CCRS, as well as CXCR2 and CXCR4. DCs respond to SDFI, but not to IL8 and MCP-I in vitro in spite of expressing CCR2 and CXCR2 mRNA. Transgenic mice, in which MCP- 1 is constitutively expressed in the epidermis under the control of the human keratin 14 promoter, respond to a contact
A. MANTOVANI
640
hypersensitivity challenge with an increased lichenoid infiltration of monocytes, T lymphocytes and CD45+, I-A+ cells that assumed a dendritic morphology in situ [ 141.We speculatethat MCP- 1 may attract monocytes to the skin, where thesecells find an environment (e.g. GM-CSF) conducive to DC differentiation.
Differential regulation during maturation
In the context of our interest in chemokines and DC, we recently transfected tumour cells with MCP-3 [ 161. After MCP-3 gene transfer, P8 15 mastocytoma cells grew, but underwent rejection. MCP-3-elicited rejection was associated with resistance to subsequent challenge with parental cells. MCP-3-elicited rejection was associated with profound alterations of leukocyte infiltration. TAMS were already present in large numbers, but T cells, eosinophils and neutrophils increased in tumour tissues after MCP-3 gene transfer. DC (e.g. Dec205+, high MHC class II+ cells) did not increase substantially in the tumour mass. However, in peritumoral tissues, DCs accumulated in perivascular areas. In contrast to their behavior, in immunocompetent mice, MCP-3-transfected tumour cells grew normally in nude mice. Increased accumulation of macrophages and PMN was also evident in nude mice. Antibodies against CD4, CD8 and IFNy, but not against IL4, inhibited rejection of MCP-3 transfected P815 cells. An anti-PMN mAb caused only a delay in MCP-3elicited tumour rejection. Thus, MCP-3 gene transfer elicits tumour rejection by activating type I T-cell-dependent immunity. It is tempting to speculate that altered trafficking of antigen-presenting cells, which express receptors and respond to MCP-3, together with recruitment of activated T cells, underlies activation of specific immunity by MCP-3-transfected cells.
= =
dendritic effective
= =
granulocyte-macrophage interferon.
Key-words: Dendritic cell, Chemokine; Receptor expression, Review. References
PI Baggiolini, M., Dewald, B. & Moser, B. (1997), Human chemokines:An update.Annu. Rev. Immu-
nol., 15,675705.
[21 Hedrick, J.A. & Zlotnik, A. (1996), Chemokinesand lymphocyte biology. Curr. Opin. Immunol., 8, 343347. r31 Rollins, B.J. (1997), Chemokines.Blood, 90, 909928. [41 Bonecchi, R., Bianchi, G., Bordignon, P.P., ef al.
(1998),Differentialexpressionof chemokinereceptors and chemotactic responsiveness of type 1 T helper cells(Thls) andTh2s.J. Exp. Med., 187, 129-134. El Sallusto, F., Mackay, C.R. & Lanzavecchia, A. (1997), Selective expressionof the eotaxin receptor CCR3 by human T helper 2 cells. Science, 277, 2005-2007.
[61 Loetscher, P., Uguccioni, M., Bordoli, L., et al. (1998), CCRS is characteristicof Thl lymphocytes. Nature, 391, 344-345. [71 Sica, A., Saccani,A., Borsatti,A., et al. (1997), Bac-
terial lipopolysacchariderapidly inhibits expression
1
cell. dose. colony-stimulating
factor.
of receptor expression
We have recently investigated how signals which induce maturation of dendritic cells affect their migration [ 17, 181.These studies were based on parallel efforts in monocytes [7, 8, 191. Maturation of DC by CD40 ligation, or culture in the presence of inflammatory agonists, such as bacterial lipopolysaccharide, IL1 and TNF, induces downregulation of the two main CC chemokine receptors expressed by these cells, CCRl and CCRS, and abrogates the chemotactic responseto their ligands, MIP- 1CI,MIPlp, RANTES and MCP-3. Inhibition of chemotaxis was rapid (< 1 h), and included the unrelated agent fMLP. Concomitantly, in the same experimental conditions, the expression of CCR7 and the migration to its ligand ELC/MIP-3/3, a chemokine expressed in lymphoid organs, was strongly upregulated, though with slower kinetics (24-48 h). Rapid inhibition of responsiveness to chemoattractants present at sites of inflammation and immune reaction may be permissive for leaving peripheral tissues. Conversely, the slower acquisition of responsiveness to ELC/MIP-3P may guide subsequent localization of DCs in lymphoid organs.
Macrophage-derived chemokine (MDC) is a recently identified CC chemokine, which, unlike related molecules, is located on chromosome 16 and has a rather distant relationship with other family members [15]. We recently found that MDC has an ED,, lOO-fold lower for activated NK cells and DC than for monocytes [ 151. Moreover, it is expressed by DC in addition to macrophages. We speculate that molecules such as SDF-1 and MDC may contribute to directing the “normal” trafficking of DC in the absence of antigen or inflammation, whereas inducible chemokines may underlie the dramatic changes in route and trafficking after exposure to antigenic or inflammatory stimulation.
DC ED GMCSF IFN
ET AL.
IL MDC PMN TAM
= = = =
interleukin. macrophage-derived polymorphonuclear tumour-associated
chemokine. (cell). macrophage.
CHEMOKINE
[8]
[9] [lo] [l l]
[12]
[ 131
[ 141
RECEPTOR
EXPRESSION
of C-C chemokine receptors in human monocytes. J. Exp. Med., 185, 969-974. Sozzani, S., Ghezzi, S., Iannolo, G. et al. (1998), Interleukin-10 increases CCRS expression and HIV infection in human monocytes. J. Exp. Med., 187, 439-444. Caux, C., Liu, Y.J. & Banchereau, J. (1995), Recent advances in the study of dendritic cells and follicular dendritic cells. Immunol. Today, 16, 2-4. Bancherau, J. & Steinman, R.M. (1998), Dendritic cells and the control of immunity. Nature, 392, 245 252. Sozzani, S., Sallusto, F., Luini, W. et al. (1995), Migration of dendritic cells in response to formyl peptides, C5a and a distinct set of chemokines. J. Immunol., 155, 3292-3295. Sallusto, F. & Lanzavecchia, A. (1994), Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/ macrophage colony-stimulating factor plus interleukin-4 and downregulated by tumor necrosis factor-alpha. J. Exp. Med., 179, 1109-1118. Piemonti, L., Bemasconi, S., Luini, W. et al. (1995), IL-13 supports differentiation of dendritic cells from circulating precursors in concert with GM-CSF. Eur. Cytokine Netw., 6, 245-252. Nakamura, K., Williams, I.R. & Kupper, T.S. (1995),
[15]
[16]
[ 171
[18]
[19]
IN DENDRITIC
CELLS
Keratinocyte-derived monocyte chemoattractant protein 1 (MCP-1): Analysis in a transgenic model demonstrates MCP-1 can recruit dendritic and langerhans cells to skin. J. Invest. Dermatol., 105, 635-643. Godiska, R., Chantry, D., Raport, C.J. etal. (1997), Human macrophage derived chemokine (MDC) a novel chemoattractant for monocytes, monocyte derived dendritic cells, and natural killer cells. J. Exp. Med., 185, 1595-1604. Fioretti, F., Fradelizi, D., Stoppacciaro, A. et al. (1998). Reduced tumorigenicity and augmented leukocyte infiltration after MCP-3 gene transfer: perivascular accumulation of dendritic cells in peritumoral tissue and neutrophil recruitment within the tumor. J. Zmmunol. (in press). Sozzani, S., Allavena, P., D’Amico, G. et al. (1998), Differential regulation of chemokine receptors during dendritic cell maturation: a model for their trafficking properties. J. Immunol. (in press). D’Amico, G., Bianchi, G., Bernasconi, S. et al. (1998), Adhesion, transendothelial migration and reverse transmigration of in vitro cultured dendritic cells. Blood (in press). Penton-Rol, G., Polentarutti, N., Luini, W. et al. (1998), Selective inhibition of expression of the chemokine receptor CCR2 in human monocytes by IFN-y. J. Immunol., 160, 3869-3873.