CD11c/CD18 Dominates Adhesion of Human Monocytes, Macrophages and Dendritic Cells over CD11b/CD18

Complement receptors CR3 (CD11b/CD18) and CR4 (CD11c/CD18) belong to the family of beta2 integrins and are expressed mainly by myeloid cell types in humans. Previously, we proved that CR3 rather than CR4 plays a key role in phagocytosis. Here we analysed how CD11b and CD11c participate in cell adhesion to fibrinogen, a common ligand of CR3 and CR4, employing human monocytes, monocyte-derived macrophages (MDMs) and monocyte-derived dendritic cells (MDDCs) highly expressing CD11b as well as CD11c. We determined the exact numbers of CD11b and CD11c on these cell types by a bead-based technique, and found that the ratio of CD11b/CD11c is 1.2 for MDDCs, 1.7 for MDMs and 7.1 for monocytes, suggesting that the function of CD11c is preponderant in MDDCs and less pronounced in monocytes. Applying state-of-the-art biophysical techniques, we proved that cellular adherence to fibrinogen is dominated by CD11c. Furthermore, we found that blocking CD11b significantly enhances the attachment of MDDCs and MDMs to fibrinogen, demonstrating a competition between CD11b and CD11c for this ligand. On the basis of the cell surface receptor numbers and the measured adhesion strength we set up a model, which explains the different behavior of the three cell types.

[1]  Robert Horvath,et al.  In-situ and label-free optical monitoring of the adhesion and spreading of primary monocytes isolated from human blood: dependence on serum concentration levels. , 2014, Biosensors & bioelectronics.

[2]  M. Sperandio,et al.  The molecular basis of leukocyte recruitment and its deficiencies. , 2013, Molecular immunology.

[3]  A. Erdei,et al.  CR 3 is the dominant phagocytotic complement receptor on human dendritic cells , 2014 .

[4]  Suet-Mien Tan,et al.  The leucocyte β2 (CD18) integrins: the structure, functional regulation and signalling properties. , 2012, Bioscience reports.

[5]  A. Prescott,et al.  Dendritic cell podosomes are protrusive and invade the extracellular matrix using metalloproteinase MMP-14 , 2010, Journal of Cell Science.

[6]  O. Destaing,et al.  Invadosome regulation by adhesion signaling. , 2011, Current opinion in cell biology.

[7]  N. Hogg,et al.  Integrins in immunity , 2009, Journal of Cell Science.

[8]  G. Schuler,et al.  Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability. , 1997, Advances in experimental medicine and biology.

[9]  Cinzia Giagulli,et al.  The Src Family Kinases Hck and Fgr Are Dispensable for Inside-Out, Chemoattractant-Induced Signaling Regulating β2 Integrin Affinity and Valency in Neutrophils, but Are Required for β2 Integrin-Mediated Outside-In Signaling Involved in Sustained Adhesion1 , 2006, The Journal of Immunology.

[10]  T. Ugarova,et al.  The role of integrin alpha D beta2 (CD11d/CD18) in monocyte/macrophage migration. , 2008, Experimental cell research.

[11]  A. Mócsai,et al.  Syk is required for integrin signaling in neutrophils. , 2002, Immunity.

[12]  J. Loike,et al.  CD11c/CD18 on neutrophils recognizes a domain at the N terminus of the A alpha chain of fibrinogen. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[13]  T. Springer,et al.  Purification and alpha subunit N-terminal sequences of human Mac-1 and p150,95 leukocyte adhesion proteins. , 1987, Journal of immunology.

[14]  T. Ugarova,et al.  Integrin alphaDbeta2, an adhesion receptor up-regulated on macrophage foam cells, exhibits multiligand-binding properties. , 2006, Blood.

[15]  P. Taylor,et al.  Phagocytosis Is the Main CR3-Mediated Function Affected by the Lupus-Associated Variant of CD11b in Human Myeloid Cells , 2013, PloS one.

[16]  F. Sallusto,et al.  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 , 1994, The Journal of experimental medicine.

[17]  T. Spector,et al.  Basic and Translational Research , 2022 .

[18]  M. Schwartz,et al.  Integrins and cell proliferation: regulation of cyclin-dependent kinases via cytoplasmic signaling pathways. , 2001, Journal of cell science.

[19]  V. Kanagasundaram,et al.  Integrin CD11c contributes to monocyte adhesion with CD11b in a differential manner and requires Src family kinase activity. , 2008, Molecular immunology.

[20]  H. Rosen,et al.  The leukocyte cell surface receptor(s) for the iC3b product of complement. , 1990, Current topics in microbiology and immunology.

[21]  J. Wautier,et al.  Blood monocyte activation in rheumatoid arthritis: increased monocyte adhesiveness, integrin expression, and cytokine release , 1996, Clinical and experimental immunology.

[22]  A. Prescott,et al.  A crucial role for β2 integrins in podosome formation, dynamics and Toll-like-receptor-signaled disassembly in dendritic cells , 2014, Journal of Cell Science.

[23]  Zsuzsanna Környei,et al.  Erratum: Cell sorting in a Petri dish controlled by computer vision (Scientific Reports) , 2013 .

[24]  T. Ugarova,et al.  Regulated Unmasking of the Cryptic Binding Site for Integrin αMβ2 in the γC-Domain of Fibrinogen† , 2002 .

[25]  D. Frommhold,et al.  Critical role of phospholipase Cγ2 in integrin and Fc receptor-mediated neutrophil functions and the effector phase of autoimmune arthritis , 2009, The Journal of experimental medicine.

[26]  D. Altieri,et al.  Differential ligand binding specificities of recombinant CD11b/CD18 integrin I-domain. , 1994, The Journal of biological chemistry.

[27]  M. Diamond,et al.  The I domain is a major recognition site on the leukocyte integrin Mac- 1 (CD11b/CD18) for four distinct adhesion ligands , 1993, The Journal of cell biology.

[28]  Péter Fürjes,et al.  Automated single cell isolation from suspension with computer vision , 2016, Scientific Reports.

[29]  M. Hristov,et al.  Differential role of monocyte subsets in atherosclerosis , 2011, Thrombosis and Haemostasis.

[30]  T. Springer,et al.  Regulated expression of the Mac-1, LFA-1, p150,95 glycoprotein family during leukocyte differentiation. , 1986, Journal of immunology.

[31]  J. Bergerat,et al.  Long term cultures of human monocytes in vitro. Impact of GM-CSF on survival and differentiation. , 1991, Journal of immunological methods.

[32]  M. Shimaoka,et al.  Exposure of acidic residues as a danger signal for recognition of fibrinogen and other macromolecules by integrin alphaXbeta2. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Loike,et al.  Complement receptor type three (CD11b/CD18) of human polymorphonuclear leukocytes recognizes fibrinogen. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[34]  F. Mach,et al.  Tumor necrosis factor-alpha (TNF-alpha) induces integrin CD11b/CD18 (Mac-1) up-regulation and migration to the CC chemokine CCL3 (MIP-1alpha) on human neutrophils through defined signalling pathways. , 2008, Cellular signalling.

[35]  Csaba Hős,et al.  Single Cell Adhesion Assay Using Computer Controlled Micropipette , 2014, PloS one.

[36]  T. Ugarova,et al.  A molecular basis for integrin alphaMbeta 2 ligand binding promiscuity. , 2002, The Journal of biological chemistry.

[37]  C. Laudanna,et al.  Generation of signals activating neutrophil functions by leukocyte integrins: LFA-1 and gp150/95, but not CR3, are able to stimulate the respiratory burst of human neutrophils , 1992, The Journal of cell biology.

[38]  M. Dierich,et al.  Cutting Edge: Productive HIV-1 Infection of Dendritic Cells via Complement Receptor Type 3 (CR3, CD11b/CD18) , 2004, The Journal of Immunology.

[39]  L. Koenderman,et al.  Acute‐phase concentrations of soluble fibrinogen inhibit neutrophil adhesion under flow conditions in vitro through interactions with ICAM‐1 and MAC‐1 (CD11b/CD18) , 2013, Journal of thrombosis and haemostasis : JTH.

[40]  A. Mócsai,et al.  Immunoreceptor-like signaling by β2 and β3 integrins , 2007 .

[41]  G. Berton.  Faculty Opinions recommendation of Critical role of phospholipase Cgamma2 in integrin and Fc receptor-mediated neutrophil functions and the effector phase of autoimmune arthritis. , 2009 .

[42]  R. Gower,et al.  CD 11 c / CD 18 Expression Is Upregulated on Blood Monocytes During Hypertriglyceridemia and Enhances Adhesion to Vascular Cell Adhesion Molecule-1 , 2010 .

[43]  P. Gregersen,et al.  Evaluation of imputation-based association in and around the integrin-alpha-M (ITGAM) gene and replication of robust association between a non-synonymous functional variant within ITGAM and systemic lupus erythematosus (SLE). , 2009, Human molecular genetics.

[44]  E. Brown,et al.  Ligand specificity of purified complement receptor type three (CD11b/CD18, alpha m beta 2, Mac-1). Indirect effects of an Arg-Gly-Asp (RGD) sequence. , 1993, Journal of immunology.

[45]  A. Mócsai,et al.  Integrin signaling in neutrophils and macrophages uses adaptors containing immunoreceptor tyrosine-based activation motifs , 2006, Nature Immunology.

[46]  A. Prescott,et al.  A Systemic Lupus Erythematosus-associated R77H Substitution in the CD11b Chain of the Mac-1 Integrin Compromises Leukocyte Adhesion and Phagocytosis* , 2011, The Journal of Biological Chemistry.

[47]  G. Ricevuti,et al.  Leukocyte CD11/CD18 integrins: biological and clinical relevance. , 1995, Haematologica.

[48]  T. Ugarova,et al.  The alternatively spliced αeC domain of human fibrinogen-420 is a novel ligand for leukocyte integrins αMβ2 and αXβ2 , 2001 .

[49]  Timothy A. Springer,et al.  Exposure of acidic residues as a danger signal for recognition of fibrinogen and other macromolecules by integrin αxβ2 , 2005 .

[50]  T. Ugarova,et al.  Integrin (cid:1) D (cid:2) 2 , an adhesion receptor up-regulated on macrophage foam cells, exhibits multiligand-binding properties , 2022 .

[51]  The alternatively spliced alpha(E)C domain of human fibrinogen-420 is a novel ligand for leukocyte integrins alpha(M)beta(2) and alpha(X)beta(2). , 2001, Blood.

[52]  A. Erdei,et al.  CR3 is the dominant phagocytotic complement receptor on human dendritic cells. , 2013, Immunobiology.

[53]  S. Linder,et al.  Tools of the trade: podosomes as multipurpose organelles of monocytic cells , 2014, Cellular and Molecular Life Sciences.

[54]  N. Hogg.  The leukocyte integrins. , 1989, Immunology today.

[55]  T. Ugarova,et al.  Regulated unmasking of the cryptic binding site for integrin alpha M beta 2 in the gamma C-domain of fibrinogen. , 2002, Biochemistry.

[56]  A. Steinkasserer,et al.  CD83 Knockdown in Monocyte-Derived Dendritic Cells by Small Interfering RNA Leads to a Diminished T Cell Stimulation1 , 2007, The Journal of Immunology.

[57]  D. Tenen,et al.  Amino acid sequence of the alpha subunit of human leukocyte adhesion receptor Mo1 (complement receptor type 3) , 1988, The Journal of cell biology.

[58]  Jeremy J. Ramsden,et al.  Dependence of cancer cell adhesion kinetics on integrin ligand surface density measured by a high-throughput label-free resonant waveguide grating biosensor , 2014, Scientific Reports.

[59]  E. Plow,et al.  A Discrete Site Modulates Activation of I Domains , 1996, The Journal of Biological Chemistry.

[60]  T. Ugarova,et al.  A Molecular Basis for Integrin αMβ2 Ligand Binding Promiscuity* , 2002, The Journal of Biological Chemistry.

[61]  C. Figdor,et al.  Role of p150,95 in adhesion, migration, chemotaxis and phagocytosis of human monocytes , 1987, European journal of immunology.

[62]  R. Gower,et al.  CD11c/CD18 Expression Is Upregulated on Blood Monocytes During Hypertriglyceridemia and Enhances Adhesion to Vascular Cell Adhesion Molecule-1 , 2011, Arteriosclerosis, thrombosis, and vascular biology.

[63]  L. Looger,et al.  Combined protein- and nucleic acid-level effects of rs1143679 (R77H), a lupus-predisposing variant within ITGAM. , 2014, Human molecular genetics.

[64]  A. Mócsai,et al.  Immunoreceptor-like signaling by beta 2 and beta 3 integrins. , 2007, Trends in cell biology.