Skip to main content
Log in

Patching and capping of LFA-1 molecules on human lymphocytes

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

The distribution and dynamics of LFA-1 molecules over the surface of human lymphocytes were analysed using immunogold label-fracture and fracture-flip methods. Patching and capping were induced by incubation at 37°C with antibodies directed against the alpha and beta chains respectively of the heterodimeric LFA-1 molecule, and were followed by immunofluorescence. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) to link LFA-1 molecules to the cytoskeleton increased the percentage of capped cells, implying a faster and more efficient process of capping. At all times of clustering or upon phorbol ester treatment, the concentration of LFA-1 in patches and then in caps was not accompained by a parallel concentration of membrane particles on the freeze-fractured plasma membranes. Our results support the role of the cytoskeleton in regulating the capping phenomenon and in controlling the structural organization of the plasma membranes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Andersson Forsman C, Pinto da Silva P (1988) Fracture-flip: new high-resolution images of cell surfaces after carbon stabilization of freeze-fractured membranes. J Cell Sci 90:531–541

    Google Scholar 

  • Bourguignon LYW, Bourguignon GJ (1984) Capping and the cytoskeleton. Int Rev Cytol 87:195–224

    Google Scholar 

  • Burn P, Kupfer A, Singer SJ (1988) Dynamic membrane-cytoskeletal interactions: specific association of integrin and talin arises in vivo after phorbol ester treatment of peripheral blood lymphocytes. Proc Natl Acad Sci USA 85:497–501

    Google Scholar 

  • Elgsaeter A, Branton D (1974) Intramembrane particle aggregation in erythrocyte ghosts. J Cell Biol 63:1018–1030

    Google Scholar 

  • Fougerolles AR de, Springer TA (1992) Intercellular adhesion molecule 3, a third adhesion counter-receptor for lymphocyte function-associated molecule 1 on resting lymphocytes. J Exp Med 175:185–190

    Google Scholar 

  • Hynes RO (1992) Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69:11–25

    Google Scholar 

  • Kammer GM, Smith JA, Mitchell R (1983) Capping of human T cell specific determinants: kinetics of capping and receptor re-expressing and regulation by the cytoskeleton. J Immunol 130:38–44

    Google Scholar 

  • Kupfer A, Singer SJ (1989) The specific interaction of helper T cells and antigen-presenting B cells. J Exp Med 170:1697–1713

    Google Scholar 

  • O'Rourke AM, Mescher MF (1990) T-cell receptor-activated adhesion systems. Curr Opin Cell Biol 2:888–893

    Google Scholar 

  • Otteskog P, Wanger L, Sundqvist KG (1983) Cytochalasins distinguish by their action resting human T lymphocytes from activated T-cell blasts. Exp Cell Res 144:443–454

    Google Scholar 

  • Pardi R, Bender JR, Dettori C, Giannazza E, Engleman EG (1989) Heterogeneous distribution and transmembrane signaling properties of lymphocyte function-associated antigens (LFA-1) in human lymphocyte subsets. J Immunol 143:3157–3166

    Google Scholar 

  • Pardi R, Inverardi L, Rugarli C, Bender JR (1992) Antigen-receptor complex stimulation triggers protein kinase C-dependent CD11a/CD18-cytoskeleton association in T lymphocytes. J Cell Biol 116:1211–1220

    Google Scholar 

  • Pavan P, Mancini P, Frati L, Torrisi MR, Pinto da Silva P (1989a) Molecular cytochemistry of CD3 and CD4 antigens in human lymphocytes as studied by label-fracture and by fracture-label. Biochim Biophys Acta 978:158–168

    Google Scholar 

  • Pavan A, Mancini P, Cirone M, Frati L, Torrisi MR, Pinto da Silva P (1989b) Capping of HLA antigens in human lymphocytes as followed by immunogold label-fracture. J Histochem Cytochem 37:1489–1496

    Google Scholar 

  • Pavan A, Mancini P, Lucania G, Frati L, Torrisi MR (1990) High-resolution surface views of human lymphocytes during capping of CD4 and HLA antigens as revealed by immunogold fracture-flip. J Cell Sci 96:151–157

    Google Scholar 

  • Pavan A, Covelli E, Pascale MC, Lucania G, Bonatti S, Pinto da Silva P, Torrisi MR (1992) Dynamics of transmembrane proteins during Sindbis virus budding. J Cell Sci 102:55–62

    Google Scholar 

  • Pinto da Silva P, Kan FWK (1984) Label-fracture: a method for high resolution labeling of cell surfaces. J Cell Biol 99:1156–1161

    Google Scholar 

  • Singer SJ (1992) Intercellular communication and cell-cell adhesion. Science 255:1671–1677

    Google Scholar 

  • Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes from immunoelectron microscopy. J Cell Biol 90:533–536

    Google Scholar 

  • Springer TA (1990) Adhesion receptors of the immune system. Nature 346:425–434

    Google Scholar 

  • Ursitti JA, Pumplin DW, Bloch RJ (1991) Ultrastructure of the human erythrocyte cytoskeleton and its attachment to the membrane. Cell Motil Cytoskel 19:227–243

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pavan, A., Lucania, G., Sansolini, T. et al. Patching and capping of LFA-1 molecules on human lymphocytes. Histochemistry 98, 253–258 (1992). https://doi.org/10.1007/BF00271039

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00271039

Keywords

Navigation