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. 1974 Mar;71(3):932–936. doi: 10.1073/pnas.71.3.932

Hybrid Antibody-Induced Topographical Redistribution of Surface Immunoglobulins, Alloantigens, and Concanavalin A Receptors on Mouse Lymphoid Cells

Christopher W Stackpole 1, Lawrence T De Milio 1, Ulrich Hämmerling 1, Janet B Jacobson 1, Michael P Lardis 1
PMCID: PMC388131  PMID: 4595577

Abstract

Redistribution of surface immunoglobulins, H-2b, Thy-1.2, and TL.1,2,3 alloantigens, and concanavalin A receptors on mouse lymphoid cells induced by hybrid rabbit F(ab′)2 antibody (anti-mouse immunoglobulin/anti-visual marker or anti-concanavalin A/anti-visual marker) was studied by immunofluorescence. When used directly to label surface immunoglobulin, and indirectly to label alloantigens and concanavalin A receptors, hybrid antibodies induced similar displacement of all surface components from a uniform distribution into “patches” and “caps” at 37°. One hybrid antibody preparation, antimouse immunoglobulin/anti-ferritin, contained negligible amounts of bivalent anti-mouse immunoglobulin antibody, and was therefore “monovalent” for the antimouse immunoglobulin specificity. This observation suggests that factors other than multivalent crosslinking are responsible for hybrid antibody-induced redistribution of cell-surface components. Cap formation induced by hybrid antibody was enhanced markedly by attachment of the visual marker, either ferritin or southern bean mosaic virus, at 37°. At -5°, hybrid antibody does not displace uniformly distributed H-2b alloantigen-alloantibody complexes, but patches of label develop when ferritin attaches to the hybrid antibody. These results explain the patchy distribution of cell-surface components, which is a temperature-independent characteristic of labeling with hybrid antibodies and visual markers for electron microscopy.

Keywords: cell surface, immunofluorescence, cap formation, ferritin

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agrawal B. B., Goldstein I. J. Protein-carbohydrate interaction. VI. Isolation of concanavalin A by specific adsorption on cross-linked dextran gels. Biochim Biophys Acta. 1967 Oct 23;147(2):262–271. [PubMed] [Google Scholar]
  2. Aoki T., Hämmerling U., De Harven E., Boyse E. A., Old L. J. Antigenic structure of cell surfaces. An immunoferritin study of the occurrence and topography of H-2' theta, and TL alloantigens on mouse cells. J Exp Med. 1969 Nov 1;130(5):979–1001. doi: 10.1084/jem.130.5.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Davis W. C., Alspaugh M. A., Stimpfling J. H., Walford R. L. Cellular surface distribution of transplantation antigens: discrepancy between direct and indirect labeling techniques. Tissue Antigens. 1971;1(2):89–93. doi: 10.1111/j.1399-0039.1971.tb00083.x. [DOI] [PubMed] [Google Scholar]
  4. Davis W. C. H-2 antigen on cell membranes: an explanation for the alteration of distribution by indirect labeling techniques. Science. 1972 Mar 3;175(4025):1006–1008. doi: 10.1126/science.175.4025.1006. [DOI] [PubMed] [Google Scholar]
  5. Edelman G. M., Yahara I., Wang J. L. Receptor mobility and receptor-cytoplasmic interactions in lymphocytes. Proc Natl Acad Sci U S A. 1973 May;70(5):1442–1446. doi: 10.1073/pnas.70.5.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frye L. D., Edidin M. The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci. 1970 Sep;7(2):319–335. doi: 10.1242/jcs.7.2.319. [DOI] [PubMed] [Google Scholar]
  7. Gingell D. Membrane permeability change by aggregation of mobile glycoprotein units. J Theor Biol. 1973 Mar;38(3):677–679. doi: 10.1016/0022-5193(73)90266-x. [DOI] [PubMed] [Google Scholar]
  8. Hämmerling U., Aoki T., Wood H. A., Old L. J., Boyse E. A., de Harvin E. New visual markers of antibody for electron microscopy. Nature. 1969 Sep 13;223(5211):1158–1159. doi: 10.1038/2231158a0. [DOI] [PubMed] [Google Scholar]
  9. Hämmerling U., Aoki T., de Harven E., Boyse E. A., Old L. J. Use of hybrid antibody with anti-gamma-G and anti-ferritin specificities in locating cell surface antigens by electron microscopy. J Exp Med. 1968 Dec 1;128(6):1461–1473. doi: 10.1084/jem.128.6.1461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hämmerling U., Rajewsky K. Evidence for surface-associated immunoglobulin on T and B lymphocytes. Eur J Immunol. 1971 Dec;1(6):447–452. doi: 10.1002/eji.1830010608. [DOI] [PubMed] [Google Scholar]
  11. Loor F., Forni L., Pernis B. The dynamic state of the lymphocyte membrane. Factors affecting the distribution and turnover of surface immunoglobulins. Eur J Immunol. 1972 Jun;2(3):203–212. doi: 10.1002/eji.1830020304. [DOI] [PubMed] [Google Scholar]
  12. Neauport-Sautes C., Lilly F., Silvestre D., Kourilsky F. M. Independence of H-2K and H-2D antigenic determinants on the surface of mouse lymphocytes. J Exp Med. 1973 Feb 1;137(2):511–526. doi: 10.1084/jem.137.2.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pinto da Silva P. Translational mobility of the membrane intercalated particles of human erythrocyte ghosts. pH-dependent, reversible aggregation. J Cell Biol. 1972 Jun;53(3):777–787. doi: 10.1083/jcb.53.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  15. Stackpole C. W., Aoki T., Boyse E. A., Old L. J., Lumley-Frank J., De Harven E. Cell surface antigens: serial sectioning of single cells as an approach to topographical analysis. Science. 1971 Apr 30;172(3982):472–474. doi: 10.1126/science.172.3982.472. [DOI] [PubMed] [Google Scholar]
  16. Tillack T. W., Scott R. E., Marchesi V. T. The structure of erythrocyte membranes studied by freeze-etching. II. Localization of receptors for phytohemagglutinin and influenza virus to the intramembranous particles. J Exp Med. 1972 Jun 1;135(6):1209–1227. doi: 10.1084/jem.135.6.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Unanue E. R., Perkins W. D., Karnovsky M. J. Ligand-induced movement of lymphocyte membrane macromolecules. I. Analysis by immunofluorescence and ultrastructural radioautography. J Exp Med. 1972 Oct 1;136(4):885–906. doi: 10.1084/jem.136.4.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yahara I., Edelman G. M. Restriction of the mobility of lymphocyte immunoglobulin receptors by concanavalin A. Proc Natl Acad Sci U S A. 1972 Mar;69(3):608–612. doi: 10.1073/pnas.69.3.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. de Petris S., Raff M. C. Distribution of immunoglobulin on the surface of mouse lymphoid cells as determined by immunoferritin electron microscopy. Antibody-induced, temperature-dependent redistribution and its implications for membrane structure. Eur J Immunol. 1972 Dec;2(6):523–535. doi: 10.1002/eji.1830020611. [DOI] [PubMed] [Google Scholar]

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