A Novel Soluble Immune-Type Receptor (SITR) in Teleost Fish: Carp SITR Is Involved in the Nitric Oxide-Mediated Response to a Protozoan Parasite

Background The innate immune system relies upon a wide range of germ-line encoded receptors including a large number of immunoglobulin superfamily (IgSF) receptors. Different Ig-like immune receptor families have been reported in mammals, birds, amphibians and fish. Most innate immune receptors of the IgSF are type I transmembrane proteins containing one or more extracellular Ig-like domains and their regulation of effector functions is mediated intracellularly by distinct stimulatory or inhibitory pathways. Methodology/Principal Findings Carp SITR was found in a substracted cDNA repertoire from carp macrophages, enriched for genes up-regulated in response to the protozoan parasite Trypanoplasma borreli. Carp SITR is a type I protein with two extracellular Ig domains in a unique organisation of a N-proximal V/C2 (or I-) type and a C-proximal V-type Ig domain, devoid of a transmembrane domain or any intracytoplasmic signalling motif. The carp SITR C-proximal V-type Ig domain, in particular, has a close sequence similarity and conserved structural characteristics to the mammalian CD300 molecules. By generating an anti-SITR antibody we could show that SITR protein expression was restricted to cells of the myeloid lineage. Carp SITR is abundantly expressed in macrophages and is secreted upon in vitro stimulation with the protozoan parasite T. borreli. Secretion of SITR protein during in vivo T. borreli infection suggests a role for this IgSF receptor in the host response to this protozoan parasite. Overexpression of carp SITR in mouse macrophages and knock-down of SITR protein expression in carp macrophages, using morpholino antisense technology, provided evidence for the involvement of carp SITR in the parasite-induced NO production. Conclusion/Significance We report the structural and functional characterization of a novel soluble immune-type receptor (SITR) in a teleost fish and propose a role for carp SITR in the NO-mediated response to a protozoan parasite.

[1]  G. Ast,et al.  Alternative splicing and evolution: diversification, exon definition and function , 2010, Nature Reviews Genetics.

[2]  H. Savelkoul,et al.  Evolution of Recognition of Ligands from Gram-Positive Bacteria: Similarities and Differences in the TLR2-Mediated Response between Mammalian Vertebrates and Teleost Fish , 2010, The Journal of Immunology.

[3]  J. Cannon,et al.  Recognition of additional roles for immunoglobulin domains in immune function. , 2010, Seminars in immunology.

[4]  X. Ju,et al.  The CD300 molecules regulate monocyte and dendritic cell functions. , 2009, Immunobiology.

[5]  A. Bateman,et al.  The evolution of protein domain families. , 2009, Biochemical Society transactions.

[6]  X. Ju,et al.  The CD300 family of molecules are evolutionarily significant regulators of leukocyte functions. , 2009, Trends in immunology.

[7]  C. Secombes,et al.  Rainbow trout (Oncorhynchus mykiss) possess multiple novel immunoglobulin-like transcripts containing either an ITAM or ITIMs. , 2009, Developmental and comparative immunology.

[8]  D. McVicar,et al.  TREM and TREM-like receptors in inflammation and disease. , 2009, Current opinion in immunology.

[9]  J. Yoder.  Form, function and phylogenetics of NITRs in bony fish. , 2009, Developmental and comparative immunology.

[10]  M. Zenke,et al.  CD300a/c regulate type I interferon and TNF-alpha secretion by human plasmacytoid dendritic cells stimulated with TLR7 and TLR9 ligands. , 2008, Blood.

[11]  M. Forlenza,et al.  Differential contribution of neutrophilic granulocytes and macrophages to nitrosative stress in a host-parasite animal model. , 2008, Molecular immunology.

[12]  P. Lenting,et al.  The Soluble Leukocyte-Associated Ig-Like Receptor (LAIR)-2 Antagonizes the Collagen/LAIR-1 Inhibitory Immune Interaction1 , 2008, The Journal of Immunology.

[13]  M. Colonna,et al.  The TREM receptor family and signal integration , 2006, Nature Immunology.

[14]  M. Joerink,et al.  Differential macrophage polarisation during parasitic infections in common carp (Cyprinus carpio L.). , 2006, Fish & shellfish immunology.

[15]  Águeda Martínez-Barriocanal,et al.  Molecular and Functional Characterization of CD300b, a New Activating Immunoglobulin Receptor Able to Transduce Signals through Two Different Pathways1 , 2006, The Journal of Immunology.

[16]  M. Joerink,et al.  Head Kidney-Derived Macrophages of Common Carp (Cyprinus carpio L.) Show Plasticity and Functional Polarization upon Differential Stimulation1 , 2006, The Journal of Immunology.

[17]  J. Lubkowski,et al.  The Structure of the Extracellular Domain of Triggering Receptor Expressed on Myeloid Cells Like Transcript-1 and Evidence for a Naturally Occurring Soluble Fragment* , 2006, Journal of Biological Chemistry.

[18]  C. Amemiya,et al.  Ancient divergence of a complex family of immune-type receptor genes , 2006, Immunogenetics.

[19]  Robert Geisler,et al.  Learning from Small Fry: The Zebrafish as a Genetic Model Organism for Aquaculture Fish Species , 2006, Marine Biotechnology.

[20]  J. Jukes,et al.  Novel immunoglobulin-like transcripts in teleost fish encode polymorphic receptors with cytoplasmic ITAM or ITIM and a new structural Ig domain similar to the natural cytotoxicity receptor NKp44 , 2005, Immunogenetics.

[21]  M. Nei,et al.  Origin and evolution of the Ig-like domains present in mammalian leukocyte receptors: insights from chicken, frog, and fish homologues , 2005, Immunogenetics.

[22]  M. Béné,et al.  A Soluble Form of the Triggering Receptor Expressed on Myeloid Cells-1 Modulates the Inflammatory Response in Murine Sepsis , 2004, The Journal of experimental medicine.

[23]  A. Órfão,et al.  Molecular Characterization of a Novel Immune Receptor Restricted to the Monocytic Lineage1 , 2004, The Journal of Immunology.

[24]  C. Amemiya,et al.  Resolution of the novel immune-type receptor gene cluster in zebrafish. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[25]  S. Levine,et al.  Mechanisms of Soluble Cytokine Receptor Generation , 2004, The Journal of Immunology.

[26]  R. Lebbink,et al.  The Mouse Homologue of the Leukocyte-Associated Ig-Like Receptor-1 Is an Inhibitory Receptor That Recruits Src Homology Region 2-Containing Protein Tyrosine Phosphatase (SHP)-2, but Not SHP-11 , 2004, The Journal of Immunology.

[27]  J. Saeij,et al.  The immune response of carp to Trypanoplasma borreli: kinetics of immune gene expression and polyclonal lymphocyte activation. , 2003, Developmental and comparative immunology.

[28]  M. Pfaffl,et al.  Standardized determination of real-time PCR efficiency from a single reaction set-up. , 2003, Nucleic acids research.

[29]  S. Beck,et al.  The human TREM gene cluster at 6p21.1 encodes both activating and inhibitory single IgV domain receptors and includes NKp44 , 2003, European journal of immunology.

[30]  A. Diefenbach,et al.  Innate immune recognition by stimulatory immunoreceptors. , 2003, Current opinion in immunology.

[31]  J. Sutcliffe,et al.  Heterogeneous expression of the triggering receptor expressed on myeloid cells‐2 on adult murine microglia , 2002, Journal of neurochemistry.

[32]  Jerzy K. Kulski,et al.  Leukocyte Ig-like receptor complex (LRC) in mice and men. , 2002, Trends in immunology.

[33]  J. Saeij,et al.  Immune modulation by fish kinetoplastid parasites: a role for nitric oxide , 2002, Parasitology.

[34]  M. Long,et al.  Evolution of novel genes. , 2001, Current opinion in genetics & development.

[35]  A. Ziegler,et al.  Genesis of the ILT/LIR/MIR clusters within the human leukocyte receptor complex , 2001, Immunological reviews.

[36]  P. Sun,et al.  Structure of killer cell immunoglobulin‐like receptors and their recognition of the class I MHC molecules , 2001, Immunological reviews.

[37]  G. Litman,et al.  Novel immune‐type receptor genes , 2001, Immunological reviews.

[38]  R. Biassoni,et al.  Human natural killer cell receptors and co‐receptors , 2001, Immunological reviews.

[39]  G. Clark,et al.  The gene encoding the immunoregulatory signaling molecule CMRF-35A localized to human chromosome 17 in close proximity to other members of the CMRF-35 family. , 2001, Tissue antigens.

[40]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[41]  Michael J. Wilson,et al.  Arrangement of the ILT gene cluster: a common null allele of the ILT6 gene results from a 6.7‐kbp deletion , 2000, European journal of immunology.

[42]  L. Lanier,et al.  Immune inhibitory receptors. , 2000, Science.

[43]  D. Steinhage,et al.  In vitro cultivation of Trypanoplasma borreli (protozoa: kinetoplastida), a parasite from the blood of common carp Cyprinus carpio. , 2000, Diseases of aquatic organisms.

[44]  G. Clark,et al.  The CMRF-35H gene structure predicts for an independently expressed member of an ITIM/ITAM pair of molecules localized to human chromosome 17. , 2000, Tissue antigens.

[45]  A. Barclay.  Ig-like domains: evolution from simple interaction molecules to sophisticated antigen recognition. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[46]  T. Wells,et al.  The Platelet Collagen Receptor Glycoprotein VI Is a Member of the Immunoglobulin Superfamily Closely Related to FcαR and the Natural Killer Receptors* , 1999, The Journal of Biological Chemistry.

[47]  L. Patthy.  Genome evolution and the evolution of exon-shuffling--a review. , 1999, Gene.

[48]  M. Hulett,et al.  Identification of residues in the first domain of human Fc alpha receptor essential for interaction with IgA. , 1999, Journal of immunology.

[49]  R. Biassoni,et al.  Molecular Cloning of NKp46: A Novel Member of the Immunoglobulin Superfamily Involved in Triggering of Natural Cytotoxicity , 1998, The Journal of experimental medicine.

[50]  S. Karlin,et al.  Finding the genes in genomic DNA. , 1998, Current opinion in structural biology.

[51]  Y. Stierhof,et al.  Cultivation of bloodstream forms of Trypanosoma carassii, a common parasite of freshwater fish , 1998, Parasitology Research.

[52]  N. Okamoto,et al.  Production of a monoclonal antibody for carp (Cyprinus carpioL.) phagocytic cells and separation of the cells , 1998 .

[53]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[54]  G. Flik,et al.  A common carp (Cyprinus carpio L) leucocyte cell line shares morphological and functional characteristics with macrophages , 1997 .

[55]  N. Taverne,et al.  Characterization of carp thrombocytes with specific monoclonal antibodies , 1996 .

[56]  Roderic D. M. Page,et al.  TreeView: an application to display phylogenetic trees on personal computers , 1996, Comput. Appl. Biosci..

[57]  D. Golde,et al.  Soluble cytokine receptors. , 1996, Blood.

[58]  P Bork,et al.  The immunoglobulin fold. Structural classification, sequence patterns and common core. , 1994, Journal of molecular biology.

[59]  B. Rinkevich,et al.  Cell adhesion receptors and nuclear receptors are highly conserved from the lowest metazoa (marine sponges) to vertebrates. , 1994, Biological chemistry Hoppe-Seyler.

[60]  C Chothia,et al.  Many of the immunoglobulin superfamily domains in cell adhesion molecules and surface receptors belong to a new structural set which is close to that containing variable domains. , 1994, Journal of molecular biology.

[61]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[62]  D. Steinhagen,et al.  The parasitemia of cloned Trypanoplasma borreli Laveran and Mesnil, 1901, in laboratory-infected common carp (Cyprinus carpio L.). , 1989, The Journal of parasitology.

[63]  D. Lipman,et al.  Improved tools for biological sequence comparison. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[64]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.

[65]  J. Felsenstein.  CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP , 1985, Evolution; international journal of organic evolution.

[66]  J. Wellink,et al.  Analysis of lymphocyte heterogeneity in carp, Cyprinus carpio L., using monoclonal antibodies , 1983 .

[67]  S. Tannenbaum,et al.  Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. , 1982, Analytical biochemistry.

[68]  L. Du Pasquier.  Speculations on the origin of the vertebrate immune system. , 2004, Immunology letters.

[69]  S. Latour,et al.  Negative regulation of immunoreceptor signaling. , 2002, Annual review of immunology.

[70]  C. Janeway,et al.  Innate immune recognition. , 2002, Annual review of immunology.

[71]  R. Biassoni,et al.  Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis. , 2001, Annual review of immunology.

[72]  E. Egberts,et al.  B cell and immunoglobulin heterogeneity in carp (Cyprinus carpio L.); an immuno(cyto)chemical study. , 1995, Developmental and comparative immunology.

[73]  I. Irnazarow.  Genetic variability of Polish and Hungarian carp lines , 1995 .

[74]  E. Egberts,et al.  Separation of lymphocyte subpopulations in carp Cyprinus carpio L. by monoclonal antibodies: immunohistochemical studies. , 1983, Immunology.