Mapping of antigenic and functional epitopes on the alpha- and beta- subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and Mac-1

Mouse Mac-1, a complement receptor-associated surface structure on macrophages, and LFA-1, a function-associated structure on lymphocytes, comprise a novel family of leukocyte differentiation antigens participating in adhesive cell interactions. Mac-1 and LFA-1 contain alpha-subunits of 170,000 and 180,000 Mr, respectively, and beta- subunits of 95,000 Mr noncovalently associated in alpha 1 beta 1 complexes. The structural relation between the alpha- and between the beta-subunits, and the location of functionally important sites on the molecules, have been probed with antibodies. Both non-cross-reactive and cross-reactive monoclonal antibodies (MAb) and antisera prepared to the purified molecules or the LFA-1 alpha-subunits were used. Reactivity with individual subunits was studied by immunoprecipitation after dissociation induced by high pH treatment, or by immunoblotting after SDS-PAGE. Cross-reactive epitopes on Mac-1 and LFA-1 were found to be present on the beta-subunits, which were immunologically identical. Non-cross-reactive epitopes that are distinctive for Mac-1 or LFA-1 were localized to the alpha-subunits. MAb to LFA-1 alpha- subunit epitopes inhibited CTL-mediated killing. Two MAb to Mac-1 alpha- subunit epitopes but not a third MAb to a spatially distinct alpha- epitope inhibited complement receptor function. Neither function was inhibited by a MAb binding to a common beta-subunit epitope. Therefore, sites of Mac-1 and LFA-1 involved in their respective adhesion-related functions, as well as distinctive structural features, have been localized to the alpha-subunits.

[1]  S. Gromkowski,et al.  The Mechanism of CTL‐Mediated Killing: Monoclonal Antibody Analysis of the Roles of Killer and Target‐Cell Membrane Proteins , 1983, Immunological reviews.

[2]  T. Springer,et al.  Biosynthesis and assembly of the alpha and beta subunits of Mac-1, a macrophage glycoprotein associated with complement receptor function. , 1983, The Journal of biological chemistry.

[3]  T. Springer,et al.  Expression and induction in vitro of macrophage differentiation antigens on murine cell lines. , 1983, Journal of immunology.

[4]  J. Strominger,et al.  Three distinct antigens associated with human T-lymphocyte-mediated cytolysis: LFA-1, LFA-2, and LFA-3. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Kelso,et al.  Clonal Heterogeneity in the Functional Requirement for Lyt‐2/3 Molecules on Cytolytic T Lymphocytes (CTL): Possible Implications for the Affinity of CTL Antigen Receptors , 1982, Immunological reviews.

[6]  Z. Eshhar,et al.  Lymphoid Cell Surface Interaction Structures Detected Using Cytolysis‐Inhibiting Monoclonal Antibodies , 1982, Immunological reviews.

[7]  F. Sánchez‐Madrid,et al.  LFA‐1 and Lyt‐2,3, Molecules Associated with T Lymphocyte‐Mediated Killing; and Mac‐1, an LFA‐1 Homologue Associated with Complement Receptor Function 1 , 1982, Immunological reviews.

[8]  Rossi Gd.  Structure and function of membrane complement receptors. Summary. , 1982 .

[9]  T. Springer,et al.  LFA-1 but not Lyt-2 is associated with killing activity of cytotoxic T lymphocyte hybridomas , 1982, Nature.

[10]  T. Springer,et al.  Purification and structural characterization of LFA-1, a lymphocyte function-associated antigen, and Mac-1, a related macrophage differentiation antigen associated with the type three complement receptor. , 1982, The Journal of biological chemistry.

[11]  R. Schreiber,et al.  Anti-mac-1 Selectively Inhibits the Mouse and Human Type Three Complement Receptor* , 2003 .

[12]  S. Wright,et al.  Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes , 1982, The Journal of experimental medicine.

[13]  F. Sánchez‐Madrid,et al.  Antigens involved in mouse cytolytic T-lymphocyte (CTL)-mediated killing: functional screening and topographic relationship. , 1982, Cellular immunology.

[14]  T. Springer,et al.  Structural homology of a macrophage differentiation antigen and an antigen involved in T-cell-mediated killing , 1982, Nature.

[15]  J. Kaufman,et al.  cDNA clones for the heavy chain of HLA-DR antigens obtained after immunopurification of polysomes by monoclonal antibody. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Lambris,et al.  Identification of a C3bi-specific membrane complement receptor that is expressed on lymphocytes, monocytes, neutrophils, and erythrocytes , 1982, The Journal of experimental medicine.

[17]  A. McMichael,et al.  Monoclonal Antibody to Human Platelet Glycoprotein I II. EFFECTS ON HUMAN PLATELET FUNCTION , 1981, British journal of haematology.

[18]  M. Omary,et al.  Molecular complexity of leukocyte surface glycoproteins related to the macrophage differentiation antigen Mac-1 , 1981, The Journal of experimental medicine.

[19]  M. Taniguchi,et al.  Mouse alloantibodies capable of blocking cytotoxic T cell function. III. Studies of the mechanism of blocking of CML by anti-Lyt-2 antibodies. , 1981, Journal of immunology.

[20]  T. Springer,et al.  Monoclonal antibody to a novel lymphocyte function-associated antigen (LFA-1): mechanism of blockade of T lymphocyte-mediated killing and effects on other T and B lymphocyte functions. , 1981, Journal of immunology.

[21]  R. Germain,et al.  A novel lymphocyte function-associated antigen (LFA-1): cellular distribution, quantitative expression, and structure. , 1981, Journal of immunology.

[22]  T. Springer,et al.  Lymphocyte function-associated antigen 1 (LFA-1): a surface antigen distinct from Lyt-2,3 that participates in T lymphocyte-mediated killing. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[23]  T. Springer.  Monoclonal antibody analysis of complex biological systems. Combination of cell hybridization and immunoadsorbents in a novel cascade procedure and its application to the macrophage cell surface. , 1981, The Journal of biological chemistry.

[24]  T. Springer,et al.  Cross-reaction of a rat-anti-mouse phagocyte-specific monoclonal antibody (anti-Mac-1) with human monocytes and natural killer cells. , 1981, Journal of immunology.

[25]  G. Berke.  Interaction of cytotoxic T lymphocytes and target cells. , 1980, Progress in allergy.

[26]  E. Reinherz,et al.  A monoclonal antibody reactive with human peripheral blood monocytes. , 1980, Journal of immunology.

[27]  H. Orr,et al.  Assembly and maturation of HLA-A and HLA-B antigens in vivo , 1979, Cell.

[28]  E. Unanue,et al.  The specific binding of Listeria monocytogenes-immune T lymphocytes to macrophages. I. Quantitation and role of H-2 gene products , 1979, The Journal of experimental medicine.

[29]  J. Michl,et al.  Effects of immobilized immune complexes on Fc- and complement-receptor function in resident and thioglycollate-elicited mouse peritoneal macrophages , 1979, The Journal of experimental medicine.

[30]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Wilson,et al.  Amino acid sequence of California quail lysozyme. Effect of evolutionary substitutions on the antigenic structure of lysozyme. , 1979, Biochemistry.

[32]  G. Galfré,et al.  Mac‐1: a macrophage differentiation antigen identified by monoclonal antibody , 1979, European journal of immunology.

[33]  A. Wilson,et al.  Further studies of an anomalous cross-reaction involving worm and vertebrate lysozymes. , 1978, Immunochemistry.

[34]  P. Fraker,et al.  Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. , 1978, Biochemical and biophysical research communications.

[35]  K. Rock,et al.  The role of H-2 linked genes in helper T-cell function. II. Isolation on antigen-pulsed macrophages of two separate populations of F1 helper T cells each specific for antigen and one set of parental H-2 products , 1978, The Journal of experimental medicine.

[36]  R A Laskey,et al.  Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film , 1977, FEBS letters.

[37]  J. Kaufman,et al.  Purification of HLA-linked B lymphocyte alloantigens in immunologically active form by preparative sodium dodecyl sulfate-gel electrophoresis and studies on their subunit association. , 1977, The Journal of biological chemistry.

[38]  S. Silverstein,et al.  Studies of the macrophage complement receptor. Alteration of receptor function upon macrophage activation , 1975, The Journal of experimental medicine.

[39]  V. Nussenzweig,et al.  RECEPTORS FOR COMPLEMENT ON LEUKOCYTES , 1968, The Journal of experimental medicine.

[40]  F. Sánchez‐Madrid,et al.  Monoclonal antibodies specific for rat IgG1, IgG2a, and IgG2b subclasses, and kappa chain monotypic and allotypic determinants: reagents for use with rat monoclonal antibodies. , 1982, Hybridoma.

[41]  C. Milstein,et al.  Preparation of monoclonal antibodies: strategies and procedures. , 1981, Methods in enzymology.

[42]  C. Milstein,et al.  [1] Preparation of monoclonal antibodies: Strategies and procedures , 1981 .

[43]  T. Springer.  Cell-Surface Differentiation in the Mouse , 1980 .

[44]  G. D. Ross.  Analysis of the different types of leukocyte membrane complement receptors and their interaction with the complement system. , 1980, Journal of immunological methods.