Specific antibody-dependent interactions between macrophages and lipid haptens in planar lipid monolayers.

Hafeman, Dean G.; Tscharner, Vinzenz von; McConnell, Harden M.

doi:10.1073/pnas.78.7.4552

Cited by 67 publications

(37 citation statements)

References 11 publications

(18 reference statements)

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“…Recent studies of the lipid monolayers on alkylated glass slides indicate that first-order transitions of the monolayers can be induced by changes in lateral pressure (25,26) and by temperature changes (unpublished data). Clearly an alkylated glass slide cannot participate in such cooperative transitions.…”

Section: Landau-de Gennes Free Energymentioning

confidence: 93%

Theory of periodic structures in lipid bilayer membranes

Falkovitz

Seul

Frisch

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

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Section: Landau-de Gennes Free Energymentioning

confidence: 93%

Theory of periodic structures in lipid bilayer membranes

Falkovitz

Seul

Frisch

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

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“…Monolayer membranes composed of dipalmitoylphosphatidylcholine (DPPC) were deposited at room temperature from an air-water interface (pressure, 30 dyn/cm) onto an alkylated glass cover slip as described previously (5,12). (Fig.…”

Section: Monolayer Membranesmentioning

confidence: 99%

Lipid monolayer-coated solid surfaces do not perturb the lateral motion and distribution of C3b receptors on neutrophils.

Hafeman¹,

Smith²,

Fearon³

et al. 1982

The Journal of Cell Biology

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We have used epifluorescence and photobleaching techniques to study the lateral distribution and motion of fluorescein-conjugated Fab fragments of anti-C3b receptor antibody bound to human neutrophils when the cells rest on various solid supports (microscope slides or cover slips). Supports composed of quartz, glass, or alkylated glass induce cellular adhesion, spreading, and an extensive lateral redistribution of C3b receptors (but not HLA antigens). The neutrophil C3b receptors become patchy, and the patches apparently undergo nonrandom translational motion. Many patches are found on the upper surfaces of the cells removed from the region of cell membrane-glass contact. In contrast, neutrophils supported by lipid monolayer-coated glass do not adhere or spread, and the C3b receptor remains uniform and diffuses freely (D = 2 x 10 -1° cm2/s).Previous studies of the lateral motion and distribution of the complement (C3b) receptor on peripheral blood leukocytes using fluorescent antireceptor antibody and antireceptor antibody fragments have indicated that these receptors are highly clustered (3, 9). Indirect measurements have also suggested that the C3b receptor is immobile on normal resting macrophages (4, 7, 8). Because cell surface receptors usually have a diffuse distribution in the absence of receptor-ligand interactions, we suspected that the clustered distribution of the C3b receptors might be a consequence of cell triggering. Concurrent studies in this laboratory have recently shown that macrophages (5), basophils (13), and neutrophils (D. G. Hafeman, M. Seul, and H. M. McConnell, unpublished observation) do not bind to lipid monolayer--coated alkylated glass surfaces in the absence of specific antibody-mediated monolayer--cell membrane interaction. We therefore undertook a study to determine whether interaction of cells with solid supports could influence the distribution and motion of the C3b receptor on human neutrophils. We find that the distribution and motion of the C3b receptor is strongly affected by the nature of the substrate on which the cells are examined. In contrast, HLA antigens are relatively unaffected by substrate contact. MATERIALS AND METHODS Preparation of CellsWhole blood was drawn from healthy human donors. Fibrin and platelets were removed by gentle swirling of the blood with 3-ram-diameter solid glass beads (one bead/ml) for 10 min. Blood was diluted with 1 vol of 0.85% NaCI. An aliquot of 6% dextran in 0.85% NaC1 (T-500; Pharmacia Fine Chemicals, Piscataway, btJ) was added to give a final dextran concentration of 0.6%. The erythrocytes were allowed to sediment at 1 g for 40 rain at 24°C. The supernatant was layered onto a cushion of Ficoll-Hypaque (Pharmacia) and granulocytes and mononuclear leukocytes were separated by centrifugation at 400 g for 40 min. Each cell fraction was washed twice in Ca +*-and Mg++-free Earle's balanced salt solution with 0.2% gelatin and buffered with 25 mM HEPES (pH 7.4). All ceils were resuspended at 5 x 10e/ml in cell buffer (CB; 2.0 mM CaCI2...

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“…Epifluorescence and total internal reflection spectroscopy have been used recently to study the interaction of cells ofthe immune system with planar phospholipid membranes (3,8,9). Investigations using optical microscopy are much more easily carried out on substrate-supported flat membranes than on vesicles and liposomes.…”

mentioning

confidence: 99%

Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates.

Subramaniam¹,

Seul²,

McConnell³

1986

Proc. Natl. Acad. Sci. U.S.A.

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We have measured the lateral mobility of fluoresceinated monoclonal IgG antibodies bound specifically to a spin label lipid hapten in phospholipid monolayers supported on alkylated silicon oxide surfaces. Dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine monolayers containing 5 mol% of the lipid hapten were transferred by conventional Langmuir-Blodgett techniques onto substrates alkylated with hydrocarbon chains containing 10, 16, and 18 carbon atoms. We show that the diffusion of the bound antibodies depends on their lateral density, the composition of the lipid monolayer, and the nature of lipid coupling to hydrocarbon chains on the alkylated substrate. Antibody diffusion coefficients at low antibody densities are within a factor of 2 of those displayed by the lipid hapten in the absence of the bound antibody. High antibody densities result in reduced antibody mobility, but the lateral diffusion of unbound lipids is unaffected.There is much current interest in the design of appropriate model systems to investigate the structure and function of cellular membranes. In recent work from our laboratory, a variety of model phospholipid membranes has been used in attempts to understand the molecular mechanisms ofcell-cell communication in the immune system (1-3). The biochemical functions of some of these cells are triggered by interactions involving antibody molecules bound to suitable receptors on cell surfaces. For example, mast cells are triggered by the crosslinking of receptor-bound IgE, and macrophages can be triggered by the binding of IgG-antigen complexes to membrane-bound F, receptors (4, 5).The specific binding and subsequent interaction of antibodies bound to lipid haptens in model phospholipid membranes is representative of a general class of events involving ligands and receptors on cell surfaces. New and improved methods for reconstitution of receptors into artificial membranes have led to much progress in understanding membrane phenomena in the immune and nervous systems (6, 7). However, many fundamental questions remain unanswered due to the unavailability of suitable model systems.Epifluorescence and total internal reflection spectroscopy have been used recently to study the interaction of cells ofthe immune system with planar phospholipid membranes (3, 8, 9). Investigations using optical microscopy are much more easily carried out on substrate-supported flat membranes than on vesicles and liposomes. Planar membranes containing histocompatibility antigens have been prepared by fusion of vesicles with solid glass substrates and employed in the study of antigen recognition by T cells (10-12). Planar monolayers and bilayers, transferred onto solid substrates from the air-water interface of a Langmuir trough, have been successfully employed as target membranes in experiments involving antibody-mediated binding of macrophages and basophils. Langmuir-Blodgett techniques afford a high degree of control over the composition and packing densities of molecules in these transferred ...

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Specific antibody-dependent interactions between macrophages and lipid haptens in planar lipid monolayers.

Cited by 67 publications

References 11 publications

Theory of periodic structures in lipid bilayer membranes

Theory of periodic structures in lipid bilayer membranes

Lipid monolayer-coated solid surfaces do not perturb the lateral motion and distribution of C3b receptors on neutrophils.

Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates.

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