Activation of Murine CD4+ and CD8+ T Lymphocytes Leads to Dramatic Remodeling of N-Linked Glycans1

Differentiation and activation of lymphocytes are documented to result in changes in glycosylation associated with biologically important consequences. In this report, we have systematically examined global changes in N-linked glycosylation following activation of murine CD4 T cells, CD8 T cells, and B cells by MALDI-TOF mass spectrometry profiling, and investigated the molecular basis for those changes by assessing alterations in the expression of glycan transferase genes. Surprisingly, the major change observed in activated CD4 and CD8 T cells was a dramatic reduction of sialylated biantennary N-glycans carrying the terminal NeuGcα2-6Gal sequence, and a corresponding increase in glycans carrying the Galα1-3Gal sequence. This change was accounted for by a decrease in the expression of the sialyltransferase ST6Gal I, and an increase in the expression of the galactosyltransferase, α1-3GalT. Conversely, in B cells no change in terminal sialylation of N-linked glycans was evident, and the expression of the same two glycosyltransferases was increased and decreased, respectively. The results have implications for differential recognition of activated and unactivated T cells by dendritic cells and B cells expressing glycan-binding proteins that recognize terminal sequences of N-linked glycans.

[1]  J. Dennis,et al.  Negative regulation of T-cell activation and autoimmunity by Mgat5 N-glycosylation , 2001, Nature.

[2]  D. Sgroi,et al.  Regulation of CD45 engagement by the B-cell receptor CD22. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Tetrahedron: Asymmetry , 2005 .

[4]  James C. Paulson,et al.  Sialoside Specificity of the Siglec Family Assessed Using Novel Multivalent Probes , 2003, Journal of Biological Chemistry.

[5]  A. Dell,et al.  Analysis of Carbohydrates/Glycoproteins by Mass Spectrometry , 2006 .

[6]  H. Pircher,et al.  Identification of an alpha2,6-sialyltransferase induced early after lymphocyte activation. , 1999, International immunology.

[7]  P. Crocker,et al.  Siglecs: sialic-acid-binding immunoglobulin-like lectins in cell-cell interactions and signalling. , 2002, Current opinion in structural biology.

[8]  A. Wagers,et al.  Potent Induction of α(1,3)-Fucosyltransferase VII in Activated CD4+ T Cells by TGF-β1 Through a p38 Mitogen-Activated Protein Kinase-Dependent Pathway1 , 2000, The Journal of Immunology.

[9]  J. Paulson,et al.  Glycomics: an integrated systems approach to structure-function relationships of glycans , 2005, Nature Methods.

[10]  J. Marth,et al.  Immune regulation by the ST6Gal sialyltransferase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Jameson,et al.  Thymocyte Sensitivity and Supramolecular Activation Cluster Formation Are Developmentally Regulated: A Partial Role for Sialylation 1 , 2003, The Journal of Immunology.

[12]  I. Stamenkovic,et al.  Natural Ligands of the B Cell Adhesion Molecule CD 22 p Carry N-Linked Oligosaccharides with ~ 2 , 6-Linked Sialic Acids That Are Required for Recognition * , 2001 .

[13]  D. Sgroi,et al.  Natural ligands of the B cell adhesion molecule CD22 beta carry N-linked oligosaccharides with alpha-2,6-linked sialic acids that are required for recognition. , 1993, The Journal of biological chemistry.

[14]  P. Shrikant,et al.  Alteration of Cell Surface Sialylation Regulates Antigen-Induced Naive CD8+ T Cell Responses1 , 2004, The Journal of Immunology.

[15]  Niclas G Karlsson,et al.  Development of a mass fingerprinting tool for automated interpretation of oligosaccharide fragmentation data , 2004, Proteomics.

[16]  A. Thall,et al.  alpha 1,3-Galactosyltransferase-deficient mice produce naturally occurring cytotoxic anti-Gal antibodies. , 1996, Transplantation proceedings.

[17]  J. Marth,et al.  The ST3Gal-I sialyltransferase controls CD8+ T lymphocyte homeostasis by modulating O-glycan biosynthesis. , 2000, Immunity.

[18]  M. Linker-Israeli,et al.  Separation of mouse thymocytes into two subpopulations by the use of peanut agglutinin. , 1976, Cellular immunology.

[19]  Mark Sutton-Smith,et al.  A rapid mass spectrometric strategy suitable for the investigation of glycan alterations in knockout mice , 2000 .

[20]  M. Fujimoto,et al.  CD22 regulates B lymphocyte function in vivo through both ligand-dependent and ligand-independent mechanisms , 2004, Nature Immunology.

[21]  D. Sgroi,et al.  The B lymphocyte adhesion molecule CD22 interacts with leukocyte common antigen CD45RO on T cells and α2–6 sialyltransferase, CD75, on B cells , 1991, Cell.

[22]  Martin Frank,et al.  Bioinformatics for glycomics: Status, methods, requirements and perspectives , 2004, Briefings Bioinform..

[23]  M. Fukuda,et al.  N-acetylglucosamine-6-O-sulfotransferases 1 and 2 cooperatively control lymphocyte homing through L-selectin ligand biosynthesis in high endothelial venules , 2005, Nature Immunology.

[24]  L. Granger,et al.  Developmentally regulated expression of peanut agglutinin (PNA)-specific glycans on murine thymocytes. , 1997, Glycobiology.

[25]  I. Wilson,et al.  Anatomy of CD1–lipid antigen complexes , 2005, Nature Reviews Immunology.

[26]  F. Luscinskas,et al.  β‐Galactoside α2,3‐sialyltransferase‐I gene expression during Th2 but not Th1 differentiation: implications for core2‐glycan formation on cell surface proteins , 2002 .

[27]  C. Dunnett A Multiple Comparison Procedure for Comparing Several Treatments with a Control , 1955 .

[28]  S. Sato,et al.  CD22, a B lymphocyte-specific adhesion molecule that regulates antigen receptor signaling. , 1997, Annual review of immunology.

[29]  Susumu Goto,et al.  Prediction of glycan structures from gene expression data based on glycosyltransferase reactions , 2005, Bioinform..

[30]  M. Fukuda,et al.  Human T-lymphocyte activation is associated with changes in O-glycan biosynthesis. , 1988, The Journal of biological chemistry.

[31]  I. Stamenkovic,et al.  Sialylation of the B lymphocyte molecule CD22 by alpha 2,6-sialyltransferase is implicated in the regulation of CD22-mediated adhesion. , 1994, The Journal of biological chemistry.

[32]  A. Wagers,et al.  Interleukin 12 and Interleukin 4 Control T Cell Adhesion to Endothelial Selectins through Opposite Effects on α1,3-fucosyltransferase VII Gene Expression , 1998, The Journal of experimental medicine.

[33]  G. Pendl,et al.  The Binding of T Cell-expressed P-selectin Glycoprotein Ligand-1 to E- and P-selectin Is Differentially Regulated* , 1997, The Journal of Biological Chemistry.

[34]  Kazuro Furukawa,et al.  Molecular Cloning of a Novel α2,3-Sialyltransferase (ST3Gal VI) That Sialylates Type II Lactosamine Structures on Glycoproteins and Glycolipids* , 1999, The Journal of Biological Chemistry.

[35]  T. Nishi,et al.  Expression Cloning of a Novel GalP( 1-3/1-4)GlcNAc cw2,3-Sialyltransferase Using Lectin Resistance Selection* , 2001 .

[36]  J. Ewing,et al.  Murine B cell differentiation is accompanied by programmed expression of multiple novel β-galactoside α2,6-sialyltransferase mRNA forms , 2000 .

[37]  K. Haas,et al.  CD22: a multifunctional receptor that regulates B lymphocyte survival and signal transduction. , 2005, Advances in immunology.

[38]  R. Kannagi,et al.  A major class of L-selectin ligands is eliminated in mice deficient in two sulfotransferases expressed in high endothelial venules , 2005, Nature Immunology.

[39]  J. Paulson,et al.  Peanut Agglutinin High Phenotype of Activated CD8+ T Cells Results from de Novo Synthesis of CD45 Glycans* , 2004, Journal of Biological Chemistry.

[40]  T. Geijtenbeek,et al.  Self- and nonself-recognition by C-type lectins on dendritic cells. , 2004, Annual review of immunology.

[41]  T. Yoshida,et al.  Cell type and maturation stage-dependent polymorphism of N-linked oligosaccharides on murine lymphocytes and lymphoma cells. , 1991, Molecular immunology.

[42]  T. J. Breen,et al.  Biostatistical Analysis (2nd ed.). , 1986 .

[43]  A. Chang,et al.  The fucosyltransferase FucT-VII regulates E-selectin ligand synthesis in human T cells , 1996, The Journal of cell biology.

[44]  L. Baum,et al.  The ST6Gal I Sialyltransferase Selectively ModifiesN-Glycans on CD45 to Negatively Regulate Galectin-1-induced CD45 Clustering, Phosphatase Modulation, and T Cell Death* , 2003, The Journal of Biological Chemistry.

[45]  J. Paulson,et al.  Regulation of alpha 2,3-sialyltransferase expression correlates with conversion of peanut agglutinin (PNA)+ to PNA- phenotype in developing thymocytes. , 1993, The Journal of biological chemistry.

[46]  S. Szabo,et al.  Molecular mechanisms regulating Th1 immune responses. , 2003, Annual review of immunology.

[47]  A. Holtorf,et al.  Variation in N-linked carbohydrate chains in different batches of two chimeric monoclonal IgG1 antibodies produced by different murine SP2/0 transfectoma cell subclones , 1995, Glycoconjugate Journal.

[48]  Hisashi Narimatsu,et al.  A focused microarray approach to functional glycomics: transcriptional regulation of the glycome. , 2006, Glycobiology.

[49]  P. Crocker Siglecs in innate immunity. , 2005, Current opinion in pharmacology.

[50]  J. E. Celis,et al.  Cell Biology: A Laboratory Handbook , 1997 .

[51]  James Paulson,et al.  Automatic annotation of matrix‐assisted laser desorption/ionization N‐glycan spectra , 2005, Proteomics.

[52]  Yoshida Tomoaki,et al.  Cell type and maturation stage-dependent polymorphism of N-linked oligosaccharides on murine lymphocytes and lymphoma cells. , 1991 .

[53]  J. Marth,et al.  Differential requirements for the O-linked branching enzyme core 2 beta1-6-N-glucosaminyltransferase in biosynthesis of ligands for E-selectin and P-selectin. , 2001, Blood.

[54]  A. Abbas,et al.  Cytokine transcriptional events during helper T cell subset differentiation , 1996, The Journal of experimental medicine.

[55]  J. Marth,et al.  Masking of CD22 by cis ligands does not prevent redistribution of CD22 to sites of cell contact. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[56]  H. Narimatsu,et al.  Expression of cutaneous lymphocyte-associated antigen regulated by a set of glycosyltransferases in human T cells: involvement of alpha1, 3-fucosyltransferase VII and beta1,4-galactosyltransferase I. , 2000, The Journal of investigative dermatology.

[57]  T. Speed,et al.  Summaries of Affymetrix GeneChip probe level data. , 2003, Nucleic acids research.

[58]  C. Janeway,et al.  α(1,3)-Fucosyltransferase VII and α(2,3)-Sialyltransferase IV Are Up-Regulated in Activated CD4 T Cells and Maintained After Their Differentiation into Th1 and Migration into Inflammatory Sites , 1999, The Journal of Immunology.

[59]  J. Marth,et al.  HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY Sialyltransferase specificity in selectin ligand formation , 2022 .

[60]  Mark E. Rogers,et al.  Sialic Acid Capping of CD8β Core 1-O-Glycans Controls Thymocyte-Major Histocompatibility Complex Class I Interaction* , 2003, The Journal of Biological Chemistry.

[61]  James C Paulson,et al.  Custom microarray for glycobiologists: considerations for glycosyltransferase gene expression profiling. , 2002, Biochemical Society symposium.

[62]  Claus-Wilhelm von der Lieth,et al.  GlycoFragment and GlycoSearchMS: web tools to support the interpretation of mass spectra of complex carbohydrates , 2004, Nucleic Acids Res..

[63]  R. Ahmed,et al.  Alterations in Cell Surface Carbohydrates on T Cells from Virally Infected Mice Can Distinguish Effector/Memory CD8+ T Cells from Naive Cells , 1998 .

[64]  R. Cummings,et al.  Glycans modulate immune responses in helminth infections and allergy. , 2006, Chemical immunology and allergy.

[65]  M. Fukuda,et al.  ST8Sia II and ST8Sia IV Polysialyltransferases Exhibit Marked Differences in Utilizing Various Acceptors Containing Oligosialic Acid and Short Polysialic Acid , 2002, The Journal of Biological Chemistry.

[66]  M. Neuberger,et al.  Interaction of CD22 with α2,6‐linked sialoglycoconjugates: innate recognition of self to dampen B cell autoreactivity? , 2002, European journal of immunology.

[67]  G. Rabinovich,et al.  Galectins as immunoregulators during infectious processes: from microbial invasion to the resolution of the disease , 2005, Parasite immunology.

[68]  Susan M. Kaech,et al.  Molecular and Functional Profiling of Memory CD8 T Cell Differentiation , 2002, Cell.

[69]  Thall Ad,et al.  alpha 1,3-Galactosyltransferase-deficient mice produce naturally occurring cytotoxic anti-Gal antibodies. , 1996 .

[70]  H. Kitagawa,et al.  Differential expression of five sialyltransferase genes in human tissues. , 1994, The Journal of biological chemistry.

[71]  D. G. Zisoulis,et al.  A crucial role for T-bet in selectin ligand expression in T helper 1 (Th1) cells. , 2005, Blood.

[72]  P. H. Atkinson,et al.  Major carbohydrate structures at five glycosylation sites on murine IgM determined by high resolution 1H-NMR spectroscopy. , 1985, Archives of biochemistry and biophysics.

[73]  Brian A. Smith,et al.  Ablation of CD22 in ligand-deficient mice restores B cell receptor signaling , 2006, Nature Immunology.

[74]  J. Lowe Glycosylation in the control of selectin counter‐receptor structure and function , 2002, Immunological reviews.

[75]  K. Sasaki,et al.  Expression cloning of a novel Gal beta (1-3/1-4) GlcNAc alpha 2,3-sialyltransferase using lectin resistance selection. , 1993, The Journal of biological chemistry.